Chemical transformations and phytochemical studies of bioactive components from extracts of Rosmarinus officinalis L
- Authors: Okoh, Omobola Oluranti
- Date: 2010
- Subjects: Essences and essential oils , Rosmarinus , Lamiaceae , Solution (Chemistry) , Extractive distillation , Medicinal plants , Bioactive compounds
- Language: English
- Type: Thesis , Doctoral , PhD (Chemistry)
- Identifier: vital:11331 , http://hdl.handle.net/10353/354 , Essences and essential oils , Rosmarinus , Lamiaceae , Solution (Chemistry) , Extractive distillation , Medicinal plants , Bioactive compounds
- Description: Variations in the yield, chemical composition, antibacterial, and antioxidant properties of the essential oils of Rosmarinus officinalis L. cultivated in Alice, Eastern Cape of South Africa over a period of 12 months using the solvent-free microwave extraction and traditional hydrodistillation methods were evaluated. The GC-MS analyses of the essential oils revealed the presence of 33 compounds with 1,8-cineole, a-pinene, camphor, verbenone, bornyl acetate and camphene constituting about 80 percent of the oils throughout the period of investigation, with the solvent-free microwave extraction method generally yielding more of the major components than the hydrodistillation method. Each of the major components of the oils varied in quantity and quality of yield at different periods of the year. The method of extraction and time of harvest are of importance to the quantity and quality of essential oil of Rosmarinus officinalis. Higher amounts of oxygenated monoterpenes such as borneol, camphor, terpene- 4-ol, linalool, a-terpeneol were present in the oil of SFME in comparison with HD. However, HD oil contained more monoterpene hydrocarbons such as a-pinene, camphene, β-pinene, myrcene, a-phellanderene, 1,8-cineole, trans- β-ocimene, γ-teprinene, and cis-sabinene hydrate than SFME extracted oil. Accumulation of monoterpene alcohols and ketones was observed during maturation process of Rosmarinus leaves. Quantitative evaluation of antibacterial activity, minimum inhibitory concentration values were determined using a serial microplate dilution method. The essential oils obtained using both methods of extraction were active against all the bacteria tested at a concentration of 10 mg mL-1. The minimum inhibitory concentrations for the SFME extracted oils ranged between 0.23 and 1.88 mg mL-1, while those of the HD extracted oils varied between 0.94 and 7.5 mg mL-1, thus suggesting that the oil obtained by solvent free microwave extraction was more active against bacteria than the oil obtained through hydrodistillation. The antioxidant and free radical scavenging activity of the obtained oils were tested by means of 1,1-diphenyl-2-picrylhydrazyl radical (DPPH+) assay and β- carotene bleaching test. In the DPPH+ assay, while the free radical scavenging activity of the oil obtained by SFME method showed percentage inhibitions of between 48.8 percent and 67 percent, the HD derived oil showed inhibitions of between 52.2 percent and 65.30 percent at concentrations of 0.33, 0.50 and 1.0 mg mL-1, respectively. In the β-carotene bleaching assay, the percentage inhibition increased with increasing concentration of both oils with a higher antioxidant activity of the oil obtained through the SFME than the HD method. Thin layer chromatography (TLC) was used to analyze the chemical composition of the extracts using three eluent solvent systems of varying polarities i. e. CEF, BEA and EMW and sprayed with vanillin-sulfuric acid. The chemical composition of the different extracts was similar with the exception of methanol and water extracts which had only one or two visible compounds after treating with vanillin-spray reagent. To evaluate the number of antibacterial compounds present in the fractions, bioautography was used against two most important nosocomial microorganisms. S. aureus (Gram positive) and E. coli (Gram negative). Nearly all the crude serial extraction fractions contained compounds that inhibited the growth of E. coli. The hexane extract had the most lines of inhibition followed by ethyl acetate. Bioassay-guided fractionation against E. coli was used to isolate antibacterial compounds. The largest number of antibacterial compounds occurred in the hexane fraction. Furthermore we tried to complete the characterization by extracting and studying other biologically important plant metabolites such as phenolic compounds to evaluate the antioxidant capacity of Rosmarinus extracts.
- Full Text:
- Date Issued: 2010
- Authors: Okoh, Omobola Oluranti
- Date: 2010
- Subjects: Essences and essential oils , Rosmarinus , Lamiaceae , Solution (Chemistry) , Extractive distillation , Medicinal plants , Bioactive compounds
- Language: English
- Type: Thesis , Doctoral , PhD (Chemistry)
- Identifier: vital:11331 , http://hdl.handle.net/10353/354 , Essences and essential oils , Rosmarinus , Lamiaceae , Solution (Chemistry) , Extractive distillation , Medicinal plants , Bioactive compounds
- Description: Variations in the yield, chemical composition, antibacterial, and antioxidant properties of the essential oils of Rosmarinus officinalis L. cultivated in Alice, Eastern Cape of South Africa over a period of 12 months using the solvent-free microwave extraction and traditional hydrodistillation methods were evaluated. The GC-MS analyses of the essential oils revealed the presence of 33 compounds with 1,8-cineole, a-pinene, camphor, verbenone, bornyl acetate and camphene constituting about 80 percent of the oils throughout the period of investigation, with the solvent-free microwave extraction method generally yielding more of the major components than the hydrodistillation method. Each of the major components of the oils varied in quantity and quality of yield at different periods of the year. The method of extraction and time of harvest are of importance to the quantity and quality of essential oil of Rosmarinus officinalis. Higher amounts of oxygenated monoterpenes such as borneol, camphor, terpene- 4-ol, linalool, a-terpeneol were present in the oil of SFME in comparison with HD. However, HD oil contained more monoterpene hydrocarbons such as a-pinene, camphene, β-pinene, myrcene, a-phellanderene, 1,8-cineole, trans- β-ocimene, γ-teprinene, and cis-sabinene hydrate than SFME extracted oil. Accumulation of monoterpene alcohols and ketones was observed during maturation process of Rosmarinus leaves. Quantitative evaluation of antibacterial activity, minimum inhibitory concentration values were determined using a serial microplate dilution method. The essential oils obtained using both methods of extraction were active against all the bacteria tested at a concentration of 10 mg mL-1. The minimum inhibitory concentrations for the SFME extracted oils ranged between 0.23 and 1.88 mg mL-1, while those of the HD extracted oils varied between 0.94 and 7.5 mg mL-1, thus suggesting that the oil obtained by solvent free microwave extraction was more active against bacteria than the oil obtained through hydrodistillation. The antioxidant and free radical scavenging activity of the obtained oils were tested by means of 1,1-diphenyl-2-picrylhydrazyl radical (DPPH+) assay and β- carotene bleaching test. In the DPPH+ assay, while the free radical scavenging activity of the oil obtained by SFME method showed percentage inhibitions of between 48.8 percent and 67 percent, the HD derived oil showed inhibitions of between 52.2 percent and 65.30 percent at concentrations of 0.33, 0.50 and 1.0 mg mL-1, respectively. In the β-carotene bleaching assay, the percentage inhibition increased with increasing concentration of both oils with a higher antioxidant activity of the oil obtained through the SFME than the HD method. Thin layer chromatography (TLC) was used to analyze the chemical composition of the extracts using three eluent solvent systems of varying polarities i. e. CEF, BEA and EMW and sprayed with vanillin-sulfuric acid. The chemical composition of the different extracts was similar with the exception of methanol and water extracts which had only one or two visible compounds after treating with vanillin-spray reagent. To evaluate the number of antibacterial compounds present in the fractions, bioautography was used against two most important nosocomial microorganisms. S. aureus (Gram positive) and E. coli (Gram negative). Nearly all the crude serial extraction fractions contained compounds that inhibited the growth of E. coli. The hexane extract had the most lines of inhibition followed by ethyl acetate. Bioassay-guided fractionation against E. coli was used to isolate antibacterial compounds. The largest number of antibacterial compounds occurred in the hexane fraction. Furthermore we tried to complete the characterization by extracting and studying other biologically important plant metabolites such as phenolic compounds to evaluate the antioxidant capacity of Rosmarinus extracts.
- Full Text:
- Date Issued: 2010
Outcomes of Drug Resistant Tuberculosis in Two Rural District Hospitals, Eastern Cape Province, South Africa
- Authors: Lotz, John-D Knipe
- Date: 2021-02
- Subjects: Medicinal plants
- Language: English
- Type: Masters theses , text
- Identifier: http://hdl.handle.net/11260/6834 , vital:51018
- Description: Tuberculosis (TB) is still rampant in South Africa, and drug resistant tuberculosis (DR-TB) forms a significant part of this burden on both the health care system and economy. A number of interventions have recently been introduced to help curb the growing epidemic of DR-TB, including increasing access to novel and repurposed drugs, decentralisation of care, and a new shorter (9-11 month) treatment regimen recently endorsed by the World Health Organization (WHO). Significantly, this new regimen has now also become injectable-free (also known as an all-oral regimen). However, at the time of implementation in 2017, the shorter regimen was yet to be proven effective in a programmatic setting in South Africa. This is a retrospective cohort study to describe the outcomes in patients on short and long DR-TB treatment regimens, over five years, at two treatment sites in a rural setting in the Eastern Cape province of South Africa. It is the hope that elucidation of factors involved in affecting outcomes in DR-TB may direct future interventions in these two facilities, and the wider DR-TB program in South Africa , Thesis (Masters) -- Faculty of Health Sciences, 2021
- Full Text:
- Date Issued: 2021-02
- Authors: Lotz, John-D Knipe
- Date: 2021-02
- Subjects: Medicinal plants
- Language: English
- Type: Masters theses , text
- Identifier: http://hdl.handle.net/11260/6834 , vital:51018
- Description: Tuberculosis (TB) is still rampant in South Africa, and drug resistant tuberculosis (DR-TB) forms a significant part of this burden on both the health care system and economy. A number of interventions have recently been introduced to help curb the growing epidemic of DR-TB, including increasing access to novel and repurposed drugs, decentralisation of care, and a new shorter (9-11 month) treatment regimen recently endorsed by the World Health Organization (WHO). Significantly, this new regimen has now also become injectable-free (also known as an all-oral regimen). However, at the time of implementation in 2017, the shorter regimen was yet to be proven effective in a programmatic setting in South Africa. This is a retrospective cohort study to describe the outcomes in patients on short and long DR-TB treatment regimens, over five years, at two treatment sites in a rural setting in the Eastern Cape province of South Africa. It is the hope that elucidation of factors involved in affecting outcomes in DR-TB may direct future interventions in these two facilities, and the wider DR-TB program in South Africa , Thesis (Masters) -- Faculty of Health Sciences, 2021
- Full Text:
- Date Issued: 2021-02
African traditional medicine-antiretroviral interactions : effects of Sutherlandia frutescens on the pharmacokinetics of Atazanavir
- Authors: Müller, Adrienne Carmel
- Date: 2011 , 2011-03-28
- Subjects: Antiretroviral agents , Medicinal plants , Traditional medicine , AIDS (Disease) -- Treatment , HIV infections -- Drug therapy , Drug interactions , Pharmacokinetics
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:3859 , http://hdl.handle.net/10962/d1013373
- Description: In response to the urgent call for investigations into antiretroviral (ARV)-African traditional medicine (ATM) interactions, this research was undertaken to ascertain whether chronic administration of the ATM, Sutherlandia frutescens (SF) may alter the bioavailability of the protease inhibitor (PI), atazanavir (ATV), which may impact on the safety or efficacy of the ARV. Prior to investigating a potential interaction between ATV and SF in vitro and in vivo, a high performance liquid chromatography method with ultraviolet detection (HPLC-UV) was developed and validated for the bioanalysis of ATV in human plasma and liver microsomes. An improved and efficient analytical method with minimal use of solvents and short run time was achieved in comparison to methods published in the literature. In addition, the method was selective, linear, accurate and precise for quantitative analysis of ATV in these studies. Molecular docking studies were conducted to compare the binding modes and affinities of ATV and two major SF constituents, Sutherlandioside B and Sutherlandin C, with the efflux transporter, P-glycoprotein (P-gp) and the CYP450 isoenzyme, CYP3A4 to determine the potential for these phytochemicals to competitively inhibit the binding of ATV to these two proteins, which are mediators of absorption and metabolism. These studies revealed that modulation of P-gp transport of ATV by Sutherlandioside B and Sutherlandin C was not likely to occur via competitive inhibition. The results further indicated that weak competitive inhibition of CYP3A4 may possibly occur in the presence of either of these two SF constituents. The Caco-2 cell line was used as an in vitro model of human intestinal absorption. Accumulation studies in these cells were conducted to ascertain whether extracts and constituents of SF have the ability to alter the absorption of ATV. The results showed that the aqueous extract of SF significantly reduced ATV accumulation, suggesting decreased ATV absorption, whilst a triterpenoid glycoside fraction isolated from SF exhibited an opposing effect. Analogous responses were elicited by the aqueous extract and a triterpenoid glycoside fraction in similar accumulation studies in P-gp overexpressing Madin–Darby Canine Kidney Strain II cells (MDCKII-MDR1), which signified that the effects of this extract and component on ATV transport in the Caco-2 cells were P-gp-mediated. The quantitative analysis of ATV in human liver microsomes after co-incubation with extracts and components of SF was conducted to determine the effects of SF on the metabolism of ATV. The aqueous and methanolic extracts of SF inhibited ATV metabolism, whilst the triterpenoid glycoside fraction had a converse effect. Analogous effects by the extracts were demonstrated in experiments conducted in CYP3A4-transfected microsomes, suggesting that the inhibition of ATV metabolism in the liver microsomes by these SF extracts was CYP3A4-mediated. A combination of Sutherlandiosides C and D also inhibited CYP3A4-mediated ATV metabolism, which was in contrast to the response elicited by the triterpenoid fraction in the liver microsomes, where other unidentified compounds, shown to be present therein, may have contributed to the activation of ATV metabolism. The in vitro studies revealed the potential for SF to alter the bioavailability of ATV, therefore a clinical study in which the effect of a multiple dose regimen of SF on the pharmacokinetics (PK) of a single dose of ATV was conducted in healthy male volunteers. The statistical analysis showed that the 90 % confidence intervals around the geometric mean ratios (ATV + SF/ATV alone) for both Cmax and AUC0-24 hours, fell well below the lower limit of the "no-effect" boundary of 0.8 – 1.25, implying that the bioavailability of ATV was significantly reduced in this cohort of subjects. It may thus be concluded that if the reduction in bioavailability observed in this clinical study is found to be clinically relevant, co-administration of SF commercial dosage forms and ATV in HIV/AIDS patients may potentially result in subtherapeutic ATV levels, which may in turn contribute to ATV resistance and/or treatment failure. This research has therefore highlighted the potential risk for toxicity or lack of efficacy of ARV regimens which may result when ATMs and PIs are used concurrently and that patients and health care practitioners alike should be aware of these perils.
- Full Text:
- Date Issued: 2011
- Authors: Müller, Adrienne Carmel
- Date: 2011 , 2011-03-28
- Subjects: Antiretroviral agents , Medicinal plants , Traditional medicine , AIDS (Disease) -- Treatment , HIV infections -- Drug therapy , Drug interactions , Pharmacokinetics
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:3859 , http://hdl.handle.net/10962/d1013373
- Description: In response to the urgent call for investigations into antiretroviral (ARV)-African traditional medicine (ATM) interactions, this research was undertaken to ascertain whether chronic administration of the ATM, Sutherlandia frutescens (SF) may alter the bioavailability of the protease inhibitor (PI), atazanavir (ATV), which may impact on the safety or efficacy of the ARV. Prior to investigating a potential interaction between ATV and SF in vitro and in vivo, a high performance liquid chromatography method with ultraviolet detection (HPLC-UV) was developed and validated for the bioanalysis of ATV in human plasma and liver microsomes. An improved and efficient analytical method with minimal use of solvents and short run time was achieved in comparison to methods published in the literature. In addition, the method was selective, linear, accurate and precise for quantitative analysis of ATV in these studies. Molecular docking studies were conducted to compare the binding modes and affinities of ATV and two major SF constituents, Sutherlandioside B and Sutherlandin C, with the efflux transporter, P-glycoprotein (P-gp) and the CYP450 isoenzyme, CYP3A4 to determine the potential for these phytochemicals to competitively inhibit the binding of ATV to these two proteins, which are mediators of absorption and metabolism. These studies revealed that modulation of P-gp transport of ATV by Sutherlandioside B and Sutherlandin C was not likely to occur via competitive inhibition. The results further indicated that weak competitive inhibition of CYP3A4 may possibly occur in the presence of either of these two SF constituents. The Caco-2 cell line was used as an in vitro model of human intestinal absorption. Accumulation studies in these cells were conducted to ascertain whether extracts and constituents of SF have the ability to alter the absorption of ATV. The results showed that the aqueous extract of SF significantly reduced ATV accumulation, suggesting decreased ATV absorption, whilst a triterpenoid glycoside fraction isolated from SF exhibited an opposing effect. Analogous responses were elicited by the aqueous extract and a triterpenoid glycoside fraction in similar accumulation studies in P-gp overexpressing Madin–Darby Canine Kidney Strain II cells (MDCKII-MDR1), which signified that the effects of this extract and component on ATV transport in the Caco-2 cells were P-gp-mediated. The quantitative analysis of ATV in human liver microsomes after co-incubation with extracts and components of SF was conducted to determine the effects of SF on the metabolism of ATV. The aqueous and methanolic extracts of SF inhibited ATV metabolism, whilst the triterpenoid glycoside fraction had a converse effect. Analogous effects by the extracts were demonstrated in experiments conducted in CYP3A4-transfected microsomes, suggesting that the inhibition of ATV metabolism in the liver microsomes by these SF extracts was CYP3A4-mediated. A combination of Sutherlandiosides C and D also inhibited CYP3A4-mediated ATV metabolism, which was in contrast to the response elicited by the triterpenoid fraction in the liver microsomes, where other unidentified compounds, shown to be present therein, may have contributed to the activation of ATV metabolism. The in vitro studies revealed the potential for SF to alter the bioavailability of ATV, therefore a clinical study in which the effect of a multiple dose regimen of SF on the pharmacokinetics (PK) of a single dose of ATV was conducted in healthy male volunteers. The statistical analysis showed that the 90 % confidence intervals around the geometric mean ratios (ATV + SF/ATV alone) for both Cmax and AUC0-24 hours, fell well below the lower limit of the "no-effect" boundary of 0.8 – 1.25, implying that the bioavailability of ATV was significantly reduced in this cohort of subjects. It may thus be concluded that if the reduction in bioavailability observed in this clinical study is found to be clinically relevant, co-administration of SF commercial dosage forms and ATV in HIV/AIDS patients may potentially result in subtherapeutic ATV levels, which may in turn contribute to ATV resistance and/or treatment failure. This research has therefore highlighted the potential risk for toxicity or lack of efficacy of ARV regimens which may result when ATMs and PIs are used concurrently and that patients and health care practitioners alike should be aware of these perils.
- Full Text:
- Date Issued: 2011
Antibacterial properties of the methanol extract of helichrysum pedunculatum
- Authors: Ncube, Nqobile S
- Date: 2008
- Subjects: Medicinal plants , Methanol , Helichrysum
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11241 , http://hdl.handle.net/10353/461 , Medicinal plants , Methanol , Helichrysum
- Description: The methanol extract of Helichrisum pedunculatum was screened for antimicrobial activity up to a concentration of 5 mg/ml using the agar dilution technique. A number of test bacterial isolates, comprising both Gram negative and Gram positive organisms were susceptible to the crude extract of the plant. The minimum inhibitory concentrations (MICs) of the extract ranged between 1 and 5 mg/ml for the susceptible organisms. The MICs of the selected antibiotics, chloramphenicol and penicillin, ranged between 2 and 4 mg/L, and 2 and 32 mg/L respectively against Bacillus cereus, Proteus vulgaris and Staphylococcus aureus OKOH1. Bactericidal activity was determined by the time kill assay. The methanol extract of the plant was not bactericidal at 1 × MIC for B. cereus, P. vulgaris and Staph. aureus OKOH1. At 2 × MIC the extract was bacteriostatic against B. cereus but bactericidal against P. vulgaris and Staph. aureus OKOH1. Combination studies were done at 1/2 × MIC, 1 × MIC and 2 × MIC of the plant extract with 1 × MIC of the antibiotics. Combinations of the plant extract and chloramphenicol resulted in mostly indifferent interactions against P. vulgaris and Staph. aureus OKOH1 but synergistic interactions at higher concentration of the plant extract for B. cereus. Penicillin combinations gave synergistic interactions at lower concentrations of the plant for P.vulgaris and Staph. aureus OKOH1 but was mostly indifferent for B. cereus.
- Full Text:
- Date Issued: 2008
- Authors: Ncube, Nqobile S
- Date: 2008
- Subjects: Medicinal plants , Methanol , Helichrysum
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11241 , http://hdl.handle.net/10353/461 , Medicinal plants , Methanol , Helichrysum
- Description: The methanol extract of Helichrisum pedunculatum was screened for antimicrobial activity up to a concentration of 5 mg/ml using the agar dilution technique. A number of test bacterial isolates, comprising both Gram negative and Gram positive organisms were susceptible to the crude extract of the plant. The minimum inhibitory concentrations (MICs) of the extract ranged between 1 and 5 mg/ml for the susceptible organisms. The MICs of the selected antibiotics, chloramphenicol and penicillin, ranged between 2 and 4 mg/L, and 2 and 32 mg/L respectively against Bacillus cereus, Proteus vulgaris and Staphylococcus aureus OKOH1. Bactericidal activity was determined by the time kill assay. The methanol extract of the plant was not bactericidal at 1 × MIC for B. cereus, P. vulgaris and Staph. aureus OKOH1. At 2 × MIC the extract was bacteriostatic against B. cereus but bactericidal against P. vulgaris and Staph. aureus OKOH1. Combination studies were done at 1/2 × MIC, 1 × MIC and 2 × MIC of the plant extract with 1 × MIC of the antibiotics. Combinations of the plant extract and chloramphenicol resulted in mostly indifferent interactions against P. vulgaris and Staph. aureus OKOH1 but synergistic interactions at higher concentration of the plant extract for B. cereus. Penicillin combinations gave synergistic interactions at lower concentrations of the plant for P.vulgaris and Staph. aureus OKOH1 but was mostly indifferent for B. cereus.
- Full Text:
- Date Issued: 2008
The effect of in vitro digestion on selected biological activities of Hypoxis sobolifera corms
- Authors: Van Rooyen, Anzel
- Date: 2013
- Subjects: Potatoes -- Therapeutic use , Medicinal plants
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10344 , http://hdl.handle.net/10948/d1020058
- Description: In South Africa part of the cultural and religious beliefs of the African people is the use of traditional remedies to treat diseases. These remedies are obtained from medicinal plants (Steenkamp, 2003). One of the most frequently traded plants in the Eastern Cape is Hypoxis, commonly known as Afrika patat, or African potato. South African traditional healers instruct patients to brew the fresh Hypoxis corm as a tea and then ingest it (Steenkamp, 2006a). This prompted an investigation into the digestive stability of a traditionally prepared Hypoxis extract. The H. sobolifera extracts were digested using a simulated gastric/small intestinal digestion and their biological activity determined. The hot water H. sobolifera extract before digestion only showed cytotoxic activity against cancer cell lines at very high concentrations which are not likely to be achieved under normal ingestion circumstances. In Chang liver cells on the other hand, chronic exposure to the hot water H. sobolifera extract increased glucose uptake in amounts similar to that of metformin. On the negative side, the glucose utilization stimulation was lost due to the simulated digestion process. The significant inhibition of AGEs by hot water H. sobolifera extract (IC50 of 6.3 Ig/ml) is a very encouraging result as treatment in the management of diabetes. This activity was only slightly reduced by the in vitro digestion process. Also observed was enzyme inhibition activity by traditionally prepared H. sobolifera, with ∝-amylase being inhibited (IC50 of approximately 250 Ig/ml) and therefore preventing or limiting starch breakdown. From the DPPH results it was clear that H. sobolifera, even when digested, is a potent anti-oxidant (IC50 of 134.4 Ig/ml when undigested compared to 162.9 when digested with β-glucosidase added to stomach digestive step). HPLC and TLC experiments revealed that rooperol which has previously been thought to be the compound responsible for the anti-oxidant activity in Hypoxis extracts, was absent from the traditional extract of H. sobolifera and therefore cannot be the sole compound exhibiting anti-oxidant activity; other compounds such as phenolics may be contributing. The phenolic and flavonoid content results revealed very highconcentrations of these compounds in the traditionally prepared H. sobolifera extract. These compounds may therefore play major roles in all of the biological activities observed from treatment with Hypoxis spp. The ROS results yielded interesting and promising results. Using standard or traditionally prepared H. sobolifera extracts, activation of differentiated U937 cells with PMA was greatly enhanced by cotreatment with the extracts, while extracts on their own did not cause significant activation. Future studies should investigate this property of the extracts as a promising immune boosterThe HPLC results showed that hypoxoside was undetectable in the hot water traditional extract and the TLC anti-oxidant experiment proved that rooperol is not present in the hot water traditional extract after treatment with β-glucosidase. This indicates that neither one of the Hypoxis compounds previously believed to be responsible for the biological activities observed are present in the extract when prepared the traditional way. Therefore, the biological activities observed in this study can be attributed to other phytochemical compounds.
- Full Text:
- Date Issued: 2013
- Authors: Van Rooyen, Anzel
- Date: 2013
- Subjects: Potatoes -- Therapeutic use , Medicinal plants
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10344 , http://hdl.handle.net/10948/d1020058
- Description: In South Africa part of the cultural and religious beliefs of the African people is the use of traditional remedies to treat diseases. These remedies are obtained from medicinal plants (Steenkamp, 2003). One of the most frequently traded plants in the Eastern Cape is Hypoxis, commonly known as Afrika patat, or African potato. South African traditional healers instruct patients to brew the fresh Hypoxis corm as a tea and then ingest it (Steenkamp, 2006a). This prompted an investigation into the digestive stability of a traditionally prepared Hypoxis extract. The H. sobolifera extracts were digested using a simulated gastric/small intestinal digestion and their biological activity determined. The hot water H. sobolifera extract before digestion only showed cytotoxic activity against cancer cell lines at very high concentrations which are not likely to be achieved under normal ingestion circumstances. In Chang liver cells on the other hand, chronic exposure to the hot water H. sobolifera extract increased glucose uptake in amounts similar to that of metformin. On the negative side, the glucose utilization stimulation was lost due to the simulated digestion process. The significant inhibition of AGEs by hot water H. sobolifera extract (IC50 of 6.3 Ig/ml) is a very encouraging result as treatment in the management of diabetes. This activity was only slightly reduced by the in vitro digestion process. Also observed was enzyme inhibition activity by traditionally prepared H. sobolifera, with ∝-amylase being inhibited (IC50 of approximately 250 Ig/ml) and therefore preventing or limiting starch breakdown. From the DPPH results it was clear that H. sobolifera, even when digested, is a potent anti-oxidant (IC50 of 134.4 Ig/ml when undigested compared to 162.9 when digested with β-glucosidase added to stomach digestive step). HPLC and TLC experiments revealed that rooperol which has previously been thought to be the compound responsible for the anti-oxidant activity in Hypoxis extracts, was absent from the traditional extract of H. sobolifera and therefore cannot be the sole compound exhibiting anti-oxidant activity; other compounds such as phenolics may be contributing. The phenolic and flavonoid content results revealed very highconcentrations of these compounds in the traditionally prepared H. sobolifera extract. These compounds may therefore play major roles in all of the biological activities observed from treatment with Hypoxis spp. The ROS results yielded interesting and promising results. Using standard or traditionally prepared H. sobolifera extracts, activation of differentiated U937 cells with PMA was greatly enhanced by cotreatment with the extracts, while extracts on their own did not cause significant activation. Future studies should investigate this property of the extracts as a promising immune boosterThe HPLC results showed that hypoxoside was undetectable in the hot water traditional extract and the TLC anti-oxidant experiment proved that rooperol is not present in the hot water traditional extract after treatment with β-glucosidase. This indicates that neither one of the Hypoxis compounds previously believed to be responsible for the biological activities observed are present in the extract when prepared the traditional way. Therefore, the biological activities observed in this study can be attributed to other phytochemical compounds.
- Full Text:
- Date Issued: 2013
Biochemical evaluation of Tulbaghia violacea harv.rhizomes in diet induced hypercholestrolemic rats
- Olorunnisola, Olubukola Sinbad
- Authors: Olorunnisola, Olubukola Sinbad
- Date: 2012
- Subjects: Violaceae , Anticoagulants (Medicine) , Antineoplastic agents , Rats , Hypercholesteremia , Cardiovascular agents , Medicinal plants
- Language: English
- Type: Thesis , Doctoral , PhD (Biochemistry)
- Identifier: vital:11273 , http://hdl.handle.net/10353/d1006900 , Violaceae , Anticoagulants (Medicine) , Antineoplastic agents , Rats , Hypercholesteremia , Cardiovascular agents , Medicinal plants
- Description: Discovery of cheap, nontoxic and readily available antiatherosclerotic drugs is an extraordinary challenge in this modern world. Atherosclerosis and cardiovascular diseases have been predicted to be the leading cause of death by the year 2030. Hence, this thesis was designed to search for plant (s) with anti-atherogenic properties, investigate its possible side effects and extrapolate its likely mechanism(s) of action. An ethnobotanical survey was employed in identification of locally important plants used for the management and treatment of cardiovascular diseases and its predisposing factors in Nkonkobe Municipality, Eastern Cape in South Africa. Information on the names of plants, their parts used and methods of preparation was collected through a questionnaire which was administered to herbalists, traditional healers and rural dwellers. The most frequently used plant (Rhizomes of Tulbaghia violacea Harv.) was investigated for toxicity using brine shrimp lethality (in vitro) and in vivo toxicity test (acute and subchronic) on rats to determine safety dosage. The in vitro antioxidant and free radical scavenging activity of the plant was investigated using models such as 1,1-diphenyl-2- picrylhydrazyl (DPPH), superoxide anions, hydrogen peroxide (H2O2), nitric oxide (NO), 2,2’- azinobis [3-ethylbenzothiazoline-6-sulfonic acid] diammonium salt (ABTS), lipid peroxidation inhibition and the ferric reducing agent. Phytochemical content and the effect of oral administration of fresh methanolic extract rhizomes of Tulbaghia violacea (250, 500 mg/kg. bwt/day) on Lipid peroxidation (TBARS), serum and tissue antioxidant enzymes in normal, hypercholesterolemic and diet induced atherogenic rats were also assessed. More so, the potential of the extract (250 and 500 mg/kg. bwt) to protect against atherogenic diet (4 percentage cholesterol 1 pecentage cholic acid and 0.5 percentage thiouracil) induced fatty streaks formation, dyslipidemia, oxidative stress and endothelial dysfunction was also investigated. Ethnobotanical study revealed that 19 plant species are used for the treatment of heart related diseases in the Municipality. 53 percentage of the plants mentioned were used for the management of chest pain, 47 percentage for high blood pressure, 42 percent for heart disease, 16 percentage for stroke and 11 percentage for the treatment of hypercholesterolemia. Tulbaghia violacea was repeatedly mentioned as the plant species used for the treatment of high blood pressure and predisposing factors in the study area. The brine shrimp cytotoxicity test revealed that fresh, dried methanolic extracts and essential oil of the T. violacea exhibited a high degree of cytotoxic activity with IC50 values of 18.18 (fresh) and 19.24 (dried) μg/ml. An IC50 value of 12. 59 μg/ml was obtained for the essential oil of the plant. The low cytotoxicity values obtained, suggested that rhizome of T. violacea may serve as a potential source of antimicrobial and anticancer agents. In vivo acute study of single oral administration of 5g/kg dose does not produce mortality or significant behavioral changes during 14 days observation. In the sub-chronic study, the extract (250, 500 mg/kg/bwt/ day) administered for a period of 28 days showed no mortality or morbidity. The weekly body and organ weight of the rats showed no significant differences between the control and the rats treated with the extract. The extract at all doses does not show any effect on of biomarkers of liver or renal damage. However, a significant decrease in the activity of ƔGT was observed in the extract treated groups. Hematological evaluation revealed that oral administration of fresh methanolic extracts of rhizomes of T. violacea does not cause anaemia or leucocytosis in the animals. Furthermore, histopathology results of the internal organs revealed no detectable inflammation. These results demonstrated that the rhizome extract of T. violacea was potentially safe for consumption orally even in chronic concentration. In vitro antioxidant evaluation showed that the essential oil, fresh and dried methanolic extracts exhibited potent antioxidant activities in a concentration dependent manner. Phytochemical investigation reveals that the fresh and the dry extract of RTV are rich in flavonoid, flavonol, phenols, tannin and proanthocyanidin, while the essential oil contained dimethy disulfide, dimethyl trisulfide, (methyl methylthio) methyl, 2,4-dithiapentane (11.35 percent) and (methylthio) acetic acid, 2- (methylthio) ethanol, 3-(methylthio) - and propanenitrile (7.20 percent). The fresh extract had higher radicals scavenging activity than the essential oil or dried extract, with 50 percentage inhibition of DPPH, hydrogen peroxide and lipid peroxidation at a concentration of 35.0 ± 0.12, 19.3 ± 0.11 and 17.9 ± 0.15 μg/ml respectively. Oral administration of methanolic extract of RTV in 125, 250 and 500 mg/kg to female Wistar rats significantly inhibited reduction of glutathione (GSH), superoxide dismutase (SOD) and catalase (CAT). The extracts also inhibited (p< 0.05) lipid peroxidation in normal, high cholesterol and diet induced atherosclerosis fed rats in a dose dependant manner. Also the extract (250 and 500 mg/kg/bwt/day) caused a significant (p<0.05) improvement in body weight of treated animals compared with untreated hypercholesterolemia control rats. The extracts also protected significantly (p<0.05) against atherogenic diet induced liver damage or fatty streaks formation in the aorta as revealed by histological examination. The anti-cholesterolemia and anti-atherosclerotic activities of the extract compared favorably well with standard drugs Gemfibrozil and Atorvastatin respectively. Conclusively, rhizomes of T. violacea possess significant anti-atherogenic activity and its mechanism of action(s) may be due to its antioxidant and anti-hypercholesterolemia properties. The results of this study also suggested that rhizome of T. violacea is relatively safe for human consumption and it may be used as an alternative to garlic.
- Full Text:
- Date Issued: 2012
- Authors: Olorunnisola, Olubukola Sinbad
- Date: 2012
- Subjects: Violaceae , Anticoagulants (Medicine) , Antineoplastic agents , Rats , Hypercholesteremia , Cardiovascular agents , Medicinal plants
- Language: English
- Type: Thesis , Doctoral , PhD (Biochemistry)
- Identifier: vital:11273 , http://hdl.handle.net/10353/d1006900 , Violaceae , Anticoagulants (Medicine) , Antineoplastic agents , Rats , Hypercholesteremia , Cardiovascular agents , Medicinal plants
- Description: Discovery of cheap, nontoxic and readily available antiatherosclerotic drugs is an extraordinary challenge in this modern world. Atherosclerosis and cardiovascular diseases have been predicted to be the leading cause of death by the year 2030. Hence, this thesis was designed to search for plant (s) with anti-atherogenic properties, investigate its possible side effects and extrapolate its likely mechanism(s) of action. An ethnobotanical survey was employed in identification of locally important plants used for the management and treatment of cardiovascular diseases and its predisposing factors in Nkonkobe Municipality, Eastern Cape in South Africa. Information on the names of plants, their parts used and methods of preparation was collected through a questionnaire which was administered to herbalists, traditional healers and rural dwellers. The most frequently used plant (Rhizomes of Tulbaghia violacea Harv.) was investigated for toxicity using brine shrimp lethality (in vitro) and in vivo toxicity test (acute and subchronic) on rats to determine safety dosage. The in vitro antioxidant and free radical scavenging activity of the plant was investigated using models such as 1,1-diphenyl-2- picrylhydrazyl (DPPH), superoxide anions, hydrogen peroxide (H2O2), nitric oxide (NO), 2,2’- azinobis [3-ethylbenzothiazoline-6-sulfonic acid] diammonium salt (ABTS), lipid peroxidation inhibition and the ferric reducing agent. Phytochemical content and the effect of oral administration of fresh methanolic extract rhizomes of Tulbaghia violacea (250, 500 mg/kg. bwt/day) on Lipid peroxidation (TBARS), serum and tissue antioxidant enzymes in normal, hypercholesterolemic and diet induced atherogenic rats were also assessed. More so, the potential of the extract (250 and 500 mg/kg. bwt) to protect against atherogenic diet (4 percentage cholesterol 1 pecentage cholic acid and 0.5 percentage thiouracil) induced fatty streaks formation, dyslipidemia, oxidative stress and endothelial dysfunction was also investigated. Ethnobotanical study revealed that 19 plant species are used for the treatment of heart related diseases in the Municipality. 53 percentage of the plants mentioned were used for the management of chest pain, 47 percentage for high blood pressure, 42 percent for heart disease, 16 percentage for stroke and 11 percentage for the treatment of hypercholesterolemia. Tulbaghia violacea was repeatedly mentioned as the plant species used for the treatment of high blood pressure and predisposing factors in the study area. The brine shrimp cytotoxicity test revealed that fresh, dried methanolic extracts and essential oil of the T. violacea exhibited a high degree of cytotoxic activity with IC50 values of 18.18 (fresh) and 19.24 (dried) μg/ml. An IC50 value of 12. 59 μg/ml was obtained for the essential oil of the plant. The low cytotoxicity values obtained, suggested that rhizome of T. violacea may serve as a potential source of antimicrobial and anticancer agents. In vivo acute study of single oral administration of 5g/kg dose does not produce mortality or significant behavioral changes during 14 days observation. In the sub-chronic study, the extract (250, 500 mg/kg/bwt/ day) administered for a period of 28 days showed no mortality or morbidity. The weekly body and organ weight of the rats showed no significant differences between the control and the rats treated with the extract. The extract at all doses does not show any effect on of biomarkers of liver or renal damage. However, a significant decrease in the activity of ƔGT was observed in the extract treated groups. Hematological evaluation revealed that oral administration of fresh methanolic extracts of rhizomes of T. violacea does not cause anaemia or leucocytosis in the animals. Furthermore, histopathology results of the internal organs revealed no detectable inflammation. These results demonstrated that the rhizome extract of T. violacea was potentially safe for consumption orally even in chronic concentration. In vitro antioxidant evaluation showed that the essential oil, fresh and dried methanolic extracts exhibited potent antioxidant activities in a concentration dependent manner. Phytochemical investigation reveals that the fresh and the dry extract of RTV are rich in flavonoid, flavonol, phenols, tannin and proanthocyanidin, while the essential oil contained dimethy disulfide, dimethyl trisulfide, (methyl methylthio) methyl, 2,4-dithiapentane (11.35 percent) and (methylthio) acetic acid, 2- (methylthio) ethanol, 3-(methylthio) - and propanenitrile (7.20 percent). The fresh extract had higher radicals scavenging activity than the essential oil or dried extract, with 50 percentage inhibition of DPPH, hydrogen peroxide and lipid peroxidation at a concentration of 35.0 ± 0.12, 19.3 ± 0.11 and 17.9 ± 0.15 μg/ml respectively. Oral administration of methanolic extract of RTV in 125, 250 and 500 mg/kg to female Wistar rats significantly inhibited reduction of glutathione (GSH), superoxide dismutase (SOD) and catalase (CAT). The extracts also inhibited (p< 0.05) lipid peroxidation in normal, high cholesterol and diet induced atherosclerosis fed rats in a dose dependant manner. Also the extract (250 and 500 mg/kg/bwt/day) caused a significant (p<0.05) improvement in body weight of treated animals compared with untreated hypercholesterolemia control rats. The extracts also protected significantly (p<0.05) against atherogenic diet induced liver damage or fatty streaks formation in the aorta as revealed by histological examination. The anti-cholesterolemia and anti-atherosclerotic activities of the extract compared favorably well with standard drugs Gemfibrozil and Atorvastatin respectively. Conclusively, rhizomes of T. violacea possess significant anti-atherogenic activity and its mechanism of action(s) may be due to its antioxidant and anti-hypercholesterolemia properties. The results of this study also suggested that rhizome of T. violacea is relatively safe for human consumption and it may be used as an alternative to garlic.
- Full Text:
- Date Issued: 2012
Medicinal properties of Moringa (Moringa Oleifera Lam) leaves and the effect of its use as a supplement on goat growth performance and meat characteristics
- Moyo, Busani https://orcid.org/0000-0001-7002-7266
- Authors: Moyo, Busani https://orcid.org/0000-0001-7002-7266
- Date: 2011-09
- Subjects: Moringa , Medicinal plants
- Language: English
- Type: Doctoral theses , text
- Identifier: http://hdl.handle.net/10353/24376 , vital:62662
- Description: The main objective of the study was to determine if feeding goats with Moringa oleifera leaves would lead to an increase in productivity and in value of the meat. The proximate, van Soet, atomic absorption spectrophotometric and soxhlet extraction methods were used to determine the nutritional value M. oleifera leaves of the South African. The in-vitro antimicrobial screening methods were used to determine antimicrobial activities M. oleifera extracts while in vitro and invivo models were used to determine the antioxidant activities of M. oleifera leaves. An evaluation of the potential of M. oleifera leaf meal as a feed supplement in terms of its effect on helminth load, goat growth performance, carcass characteristics, meat quality attributes, nutritional and consumer sensory characteristics of goat meat was done. A total of 24, eight month old goats were randomly allocated to dietary treatments of M. oleifera leaf meal (MOL), sunflower seed cake (SC) and GH (grass hay) which was the control. All the groups were fed on basal diet of grass hay ad libitum and 200g wheat bran per head per day. The MOL group was given an additional 200 g of dried M. oleifera leaves while the SC group was offered 170 g sunflower seed cake per head/day. The study showed that the dried leaves had crude protein levels of 30.3 percent, polyunsaturated fatty acids (52.21 percent), Saturated fatty acids (43.31), n-3 (44.57 percent), n-6 (7.64 percent), 19 amino acids, vitamin E (77 mg/100 g) and Beta-carotene (18.5 mg/100 g). The M. oleifera leaf extracts showed antibacterial activities against Escherichia coli, Enterobacter cloace, Proteus vulgaris, Staphylococcus aureus and Micrococcus kristinae. The supplementation of goats with MOL and SC resulted in decreased feacal larval count and lower Haemonchus contortus, Trichostrongylus colubriforms and Oesophagastum columbianum worm burdens than those in the non-supplemented goats. Goats supplemented with SC and MOL had higher average daily weight gain and heavier carcasses than those in the GH group. Higher pH1 scores were observed in chevon from GH diet than the supplemented ones. The MOL and SC supplemented goats had chevon with higher values for lightness (L*) 24 hr post-mortem than the one from the GH group. The redness (a*) values of chevon 24 hr post mortem was highest in MOL supplemented goats. Warner Bratzler shear force (WBSF) values of SC (30.1 N) and MOL (29.8 N) supplemented goats were lower than those from GH diet (32.6 N). Chevon from goats fed GH diet had significantly higher cooking losses (29.5 percent) than that from MOL (25.4 percent) and SC (25.6 percent) fed groups. It was observed that chevon from MOL and SC supplemented groups had higher crude protein (23.57 and 22.95 percent, respectively) than the one from the GH group (21.20 percent). Cholesterol levels were higher in chevon from SC (42.84) supplemented goats than those from MOL (38.76) and GH (35.63 mg). Chevon from GH and MOL group had higher (P < 0.05) proportions of PUFA, n-3, PUFA/SFA ratio and lower n-6/n-3 ratio. Mean consumer scores for first bite, aroma, flavour and juiceness were higher in the MOL group than in the GH group (P < 0.05). The acetone extract exhibited higher concentrations of total flavonoids, flavonols, phenolics. The acetone extracts depicted higher percentage inhibition against DPPH, ABTS and nitric oxide radicals which were comparable with reference antioxidant (vitamin C and BHT). The M. oleifera leaf meal increased the antioxidant activity of GSH, SOD and catalase. Moringa oleifera leaves also exhibited medicinal properties by having anthelmintic, antibacterial activities and showed antioxidant properties. It was also observed that protein supplementation improved the animal growth performance, the physico-chemical characteristics, nutritional and fatty acids composition of meat hence meeting the consumer needs. , Thesis (PhD) -- Faculty of Science and Agriculture, 2011
- Full Text:
- Date Issued: 2011-09
- Authors: Moyo, Busani https://orcid.org/0000-0001-7002-7266
- Date: 2011-09
- Subjects: Moringa , Medicinal plants
- Language: English
- Type: Doctoral theses , text
- Identifier: http://hdl.handle.net/10353/24376 , vital:62662
- Description: The main objective of the study was to determine if feeding goats with Moringa oleifera leaves would lead to an increase in productivity and in value of the meat. The proximate, van Soet, atomic absorption spectrophotometric and soxhlet extraction methods were used to determine the nutritional value M. oleifera leaves of the South African. The in-vitro antimicrobial screening methods were used to determine antimicrobial activities M. oleifera extracts while in vitro and invivo models were used to determine the antioxidant activities of M. oleifera leaves. An evaluation of the potential of M. oleifera leaf meal as a feed supplement in terms of its effect on helminth load, goat growth performance, carcass characteristics, meat quality attributes, nutritional and consumer sensory characteristics of goat meat was done. A total of 24, eight month old goats were randomly allocated to dietary treatments of M. oleifera leaf meal (MOL), sunflower seed cake (SC) and GH (grass hay) which was the control. All the groups were fed on basal diet of grass hay ad libitum and 200g wheat bran per head per day. The MOL group was given an additional 200 g of dried M. oleifera leaves while the SC group was offered 170 g sunflower seed cake per head/day. The study showed that the dried leaves had crude protein levels of 30.3 percent, polyunsaturated fatty acids (52.21 percent), Saturated fatty acids (43.31), n-3 (44.57 percent), n-6 (7.64 percent), 19 amino acids, vitamin E (77 mg/100 g) and Beta-carotene (18.5 mg/100 g). The M. oleifera leaf extracts showed antibacterial activities against Escherichia coli, Enterobacter cloace, Proteus vulgaris, Staphylococcus aureus and Micrococcus kristinae. The supplementation of goats with MOL and SC resulted in decreased feacal larval count and lower Haemonchus contortus, Trichostrongylus colubriforms and Oesophagastum columbianum worm burdens than those in the non-supplemented goats. Goats supplemented with SC and MOL had higher average daily weight gain and heavier carcasses than those in the GH group. Higher pH1 scores were observed in chevon from GH diet than the supplemented ones. The MOL and SC supplemented goats had chevon with higher values for lightness (L*) 24 hr post-mortem than the one from the GH group. The redness (a*) values of chevon 24 hr post mortem was highest in MOL supplemented goats. Warner Bratzler shear force (WBSF) values of SC (30.1 N) and MOL (29.8 N) supplemented goats were lower than those from GH diet (32.6 N). Chevon from goats fed GH diet had significantly higher cooking losses (29.5 percent) than that from MOL (25.4 percent) and SC (25.6 percent) fed groups. It was observed that chevon from MOL and SC supplemented groups had higher crude protein (23.57 and 22.95 percent, respectively) than the one from the GH group (21.20 percent). Cholesterol levels were higher in chevon from SC (42.84) supplemented goats than those from MOL (38.76) and GH (35.63 mg). Chevon from GH and MOL group had higher (P < 0.05) proportions of PUFA, n-3, PUFA/SFA ratio and lower n-6/n-3 ratio. Mean consumer scores for first bite, aroma, flavour and juiceness were higher in the MOL group than in the GH group (P < 0.05). The acetone extract exhibited higher concentrations of total flavonoids, flavonols, phenolics. The acetone extracts depicted higher percentage inhibition against DPPH, ABTS and nitric oxide radicals which were comparable with reference antioxidant (vitamin C and BHT). The M. oleifera leaf meal increased the antioxidant activity of GSH, SOD and catalase. Moringa oleifera leaves also exhibited medicinal properties by having anthelmintic, antibacterial activities and showed antioxidant properties. It was also observed that protein supplementation improved the animal growth performance, the physico-chemical characteristics, nutritional and fatty acids composition of meat hence meeting the consumer needs. , Thesis (PhD) -- Faculty of Science and Agriculture, 2011
- Full Text:
- Date Issued: 2011-09
The medicinal plant Sutherlandia Frutescens regulates gene expression to reverse insulin resistace in rats
- Authors: Fortuin, Melissa
- Date: 2013
- Subjects: Insulin resistance , Medicinal plants , Genetic regulation , Insulin resistance -- Animal models
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10349 , http://hdl.handle.net/10948/d1020823
- Description: Obesity can lead to Type 2 Diabetes, both conditions increase in association with physical inactivity and high-energy diets, resulting in elevated blood glucose, decreased insulin sensitivity and increased insulin resistance. Sutherlandia frutescens (S.frutescens), an anti-diabetic plant, reverses and prevents insulin resistance in a rat model and human cell culture model. Gene expression analysis in hepatocyte cultures, identified genes down regulated in insulin resistance and up regulated by S.frutescens. These included genes encoding vesicle transporter proteins, hypothesised to be linked to hepatic lipid accumulation and lipid droplet formation during insulin resistance. The aim of this study was to investigate critical genes involved in lipid droplet formation, vesicle assembly and transport in high fat diet (HFD)-induced insulin resistant rat liver tissue during the development of insulin resistance and the reversal of these changes by S.frutescens. Rats were fed a low fat diet (LFD) or HFD supplemented with S.frutescens for 2, 4 and 8 weeks. Rats fed a HFD for 12 weeks developed insulin resistance, confirmed by plasma glucose and insulin levels (compared to normal controls). Groups of these rats were gavaged with S. frutescens (50mg/kg BW), Metformin (13mg/kg BW) or water for a further 4 weeks and starved for 12 hours, anaesthetized and blood removed by heart puncture. Liver was stored in RNA-Later™ for qRT-PCR and snap-frozen in liquid nitrogen for western blotting and confocal microscopy analysis. Changes in expression of vesicle transporter genes VAMP3 and NSF were analysed by qRT-PCR and changes in the protein expression by western blotting analysis. Proteins were localised within the liver by confocal immunohistochemistry using ZEN lite™ software. Statistical analysis was performed using One-Way ANOVA and unpaired t-test. mRNA gene expression of vesicle transport components VAMP3, NSF and SNAP25 showed relatively moderate changes with considerable individual variation within control or experimental groups. Uncorrelated changes in mRNA and protein products were found and may be due to differential regulation by siRNA. Proteins also showed altered staining patterns in high fat diet rats that reverted towards normal on S. frutescens treatment, potentially reflecting functional changes associated with transport of lipid-filled vesicles.
- Full Text:
- Date Issued: 2013
- Authors: Fortuin, Melissa
- Date: 2013
- Subjects: Insulin resistance , Medicinal plants , Genetic regulation , Insulin resistance -- Animal models
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10349 , http://hdl.handle.net/10948/d1020823
- Description: Obesity can lead to Type 2 Diabetes, both conditions increase in association with physical inactivity and high-energy diets, resulting in elevated blood glucose, decreased insulin sensitivity and increased insulin resistance. Sutherlandia frutescens (S.frutescens), an anti-diabetic plant, reverses and prevents insulin resistance in a rat model and human cell culture model. Gene expression analysis in hepatocyte cultures, identified genes down regulated in insulin resistance and up regulated by S.frutescens. These included genes encoding vesicle transporter proteins, hypothesised to be linked to hepatic lipid accumulation and lipid droplet formation during insulin resistance. The aim of this study was to investigate critical genes involved in lipid droplet formation, vesicle assembly and transport in high fat diet (HFD)-induced insulin resistant rat liver tissue during the development of insulin resistance and the reversal of these changes by S.frutescens. Rats were fed a low fat diet (LFD) or HFD supplemented with S.frutescens for 2, 4 and 8 weeks. Rats fed a HFD for 12 weeks developed insulin resistance, confirmed by plasma glucose and insulin levels (compared to normal controls). Groups of these rats were gavaged with S. frutescens (50mg/kg BW), Metformin (13mg/kg BW) or water for a further 4 weeks and starved for 12 hours, anaesthetized and blood removed by heart puncture. Liver was stored in RNA-Later™ for qRT-PCR and snap-frozen in liquid nitrogen for western blotting and confocal microscopy analysis. Changes in expression of vesicle transporter genes VAMP3 and NSF were analysed by qRT-PCR and changes in the protein expression by western blotting analysis. Proteins were localised within the liver by confocal immunohistochemistry using ZEN lite™ software. Statistical analysis was performed using One-Way ANOVA and unpaired t-test. mRNA gene expression of vesicle transport components VAMP3, NSF and SNAP25 showed relatively moderate changes with considerable individual variation within control or experimental groups. Uncorrelated changes in mRNA and protein products were found and may be due to differential regulation by siRNA. Proteins also showed altered staining patterns in high fat diet rats that reverted towards normal on S. frutescens treatment, potentially reflecting functional changes associated with transport of lipid-filled vesicles.
- Full Text:
- Date Issued: 2013
Phytochemical analysis and bioactivity of Garcinia Kola (Heckel) seeds on selected bacterial pathogens
- Seanego, Christinah Tshephisho
- Authors: Seanego, Christinah Tshephisho
- Date: 2012
- Subjects: Drug resistance in microorganisms , Garcinia , Antibiotics , Medicinal plants , Microbial sensitivity tests , Streptococcal infections , Streptococcus , Staphylococcus aureus infections , Salmonella typhimurium , Traditional medicine
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11259 , http://hdl.handle.net/10353/420 , Drug resistance in microorganisms , Garcinia , Antibiotics , Medicinal plants , Microbial sensitivity tests , Streptococcal infections , Streptococcus , Staphylococcus aureus infections , Salmonella typhimurium , Traditional medicine
- Description: Garcinia kola is one of the plants used in folklore remedies for the treatment of microbial infections. Bacterial resistance to commonly used antibiotics has necessitated the search for newer and alternative compounds for the treatment of drug resistant microbial infections. This study focuses on the bioactivity of G. kola seeds on Streptococcus pyogenes (ATCC 49399), Staphylococcus aureus (NCTC 6571), Plesiomonas Shigelloides (ATCC 51903) and Salmonella typhimurium (ATCC 13311), organisms which can cause illnesses from mild to severe with potentially fatal outcomes. The crude ethyl acetate, ethanol, methanol, acetone and aqueous extracts were screened by agar-well diffusion method and the activities of the extract were further determined by Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) assays. The inhibition zones ranged from 0 - 24 mm, while MIC and MBC of the extract ranged between 0.04 - 1.25 mg/mL and 0.081 - 2.5 mg/mL respectively. Chloroform/ Ethyl Acetate/ Formic acid (CEF) solvent system separated more active compounds followed by Ethyl Acetate/ Methanol/ Water (EMW) and Benzene/ Ethanol/ Ammonium Hydroxide (BEA). The extracts were fractionated by Thin Layer Chromatography (TLC). Bioautography was used to assess the activity of the possible classes of compounds present in the more active extracts. Column chromatography was used to purify the active compounds from the mixture while Gas Chromatography-Mass Spectrometry (GC-MS) was used to identify the phyto components of the fractions. The MIC of the fractions ranged between 0.0006 - 2.5 mg/mL. CEF 3 (F3), CEF 11 (F11) and CEF 12 (F12) revealed the presence of high levels fatty acids Linoleic acid, 1, 2-Benzenedicarboxylic acid and 2, 3-Dihydro-3, 5-dihydroxy-6-methyl, respectively. The results obtained from this study justify the use of this plant in traditional medicine and provide leads which could be further exploited for the development of new and potent antimicrobials.
- Full Text:
- Date Issued: 2012
- Authors: Seanego, Christinah Tshephisho
- Date: 2012
- Subjects: Drug resistance in microorganisms , Garcinia , Antibiotics , Medicinal plants , Microbial sensitivity tests , Streptococcal infections , Streptococcus , Staphylococcus aureus infections , Salmonella typhimurium , Traditional medicine
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11259 , http://hdl.handle.net/10353/420 , Drug resistance in microorganisms , Garcinia , Antibiotics , Medicinal plants , Microbial sensitivity tests , Streptococcal infections , Streptococcus , Staphylococcus aureus infections , Salmonella typhimurium , Traditional medicine
- Description: Garcinia kola is one of the plants used in folklore remedies for the treatment of microbial infections. Bacterial resistance to commonly used antibiotics has necessitated the search for newer and alternative compounds for the treatment of drug resistant microbial infections. This study focuses on the bioactivity of G. kola seeds on Streptococcus pyogenes (ATCC 49399), Staphylococcus aureus (NCTC 6571), Plesiomonas Shigelloides (ATCC 51903) and Salmonella typhimurium (ATCC 13311), organisms which can cause illnesses from mild to severe with potentially fatal outcomes. The crude ethyl acetate, ethanol, methanol, acetone and aqueous extracts were screened by agar-well diffusion method and the activities of the extract were further determined by Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) assays. The inhibition zones ranged from 0 - 24 mm, while MIC and MBC of the extract ranged between 0.04 - 1.25 mg/mL and 0.081 - 2.5 mg/mL respectively. Chloroform/ Ethyl Acetate/ Formic acid (CEF) solvent system separated more active compounds followed by Ethyl Acetate/ Methanol/ Water (EMW) and Benzene/ Ethanol/ Ammonium Hydroxide (BEA). The extracts were fractionated by Thin Layer Chromatography (TLC). Bioautography was used to assess the activity of the possible classes of compounds present in the more active extracts. Column chromatography was used to purify the active compounds from the mixture while Gas Chromatography-Mass Spectrometry (GC-MS) was used to identify the phyto components of the fractions. The MIC of the fractions ranged between 0.0006 - 2.5 mg/mL. CEF 3 (F3), CEF 11 (F11) and CEF 12 (F12) revealed the presence of high levels fatty acids Linoleic acid, 1, 2-Benzenedicarboxylic acid and 2, 3-Dihydro-3, 5-dihydroxy-6-methyl, respectively. The results obtained from this study justify the use of this plant in traditional medicine and provide leads which could be further exploited for the development of new and potent antimicrobials.
- Full Text:
- Date Issued: 2012
The in vitro biological activities of three Hypoxis species and their active compounds
- Authors: Boukes, Gerhardt Johannes
- Date: 2010
- Subjects: Potatoes -- Africa , Potatoes -- Therapeutic use , Medicinal plants
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:10322 , http://hdl.handle.net/10948/1228 , Potatoes -- Africa , Potatoes -- Therapeutic use , Medicinal plants
- Description: The African potato is used as an African traditional medicine for its nutritional and medicinal properties. Most research has been carried out on H. hemerocallidea, with very little or nothing on other Hypoxis spp. The main aim of this project was to provide scientific data on the anticancer, anti-inflammatory and antioxidant properties of H. hemerocallidea, H. stellipilis and H. sobolifera chloroform extracts and their active compounds. The hypoxoside and phytosterol contents of the three Hypoxis spp. were determined using TLC, HPLC and GC. H. hemerocallidea and H. sobolifera chloroform extracts contained the highest amounts of hypoxoside and β-sitosterol, respectively. For the anticancer properties, cytotoxicity of the Hypoxis extracts and its purified compounds were determined against the HeLa, HT-29 and MCF-7 cancer cell lines (using MTT), and PBMCs (using CellTiter-Blue®). H. sobolifera had the best cytotoxicity against the three cancer cell lines, whereas H. stellipilis stimulated HeLa and HT-29 cancer cell growth. IC50 values of hypoxoside and rooperol were determined. DNA cell cycle arrest (using PI staining) occurred in the late G1/early S (confirmed by increased p21Waf1/Cip1 expression) and G2/M phases after 15 and 48 hrs, respectively, when treated with Hypoxis extracts and rooperol. H. sobolifera and rooperol activated caspase-3 and -7 (using fluorescently labelled antibodies) in HeLa and HT-29 cancer cells, and caspase-7 in MCF-7 cancer cells after 48 hrs. Annexin V binding to phosphatidylserines in rooperol treated U937 cells confirmed early apoptosis after 15 hrs. The TUNEL assay showed DNA fragmentation in the three cancer cell lines when treated with H. sobolifera and rooperol for 48 hrs. A shift pass the G2/M phase has led to the investigation of endoreduplication, which was confirmed by cell/nucleus size, and anti-apoptotic proteins (Akt, phospho-Akt, phospho-Bcl-2 and p21Waf1/Cip1). U937 cell differentiation to monocyte-macrophages was optimized using PMA and 1,25(OH)2D3, which was confirmed by morphological and biochemical changes. For the anti-inflammatory properties, Hypoxis extracts and rooperol significantly increased NO production in monocyte-macrophages (pre-loaded with DAF-2 DA) and phagocytosis of pHrodoTM E. coli BioParticles®. The treatments had no effect on COX-2 expression in monocyte-macrophages. The phytosterols significantly increased IL-1β and IL-6 secretion xv (using the FlowCytomix Multiplex human Th1/Th2 10plex Kit I) in the PBMCs of one donor. For the antioxidant properties, Hypoxis extracts and rooperol significantly increased ROS production in undifferentiated and differentiated U937 cells, which were pre-loaded with DCFH-DA. Hypoxis extracts and purified compounds had ferric reducing activities, but only rooperol had ferric reducing activities significantly greater than ascorbic acid. β-sitosterol, campesterol and cholesterol significantly increased SOD activity in Chang liver cells, while H. stellipilis, H. sobolifera and rooperol decreased SOD activity. Anticancer, anti-inflammatory and antioxidant properties of the Hypoxis extracts may be attributed to the β-sitosterol content, because Hypoxis chloroform extracts contained very little or no hypoxoside. Unidentified compounds, and synergistic and additive effects of the compounds may have contributed to the biological effects. This study confirms previous reports that rooperol is the active compound. Results provide scientific data on the medicinal properties of one of the most frequently used medicinal plants in South Africa.
- Full Text:
- Date Issued: 2010
- Authors: Boukes, Gerhardt Johannes
- Date: 2010
- Subjects: Potatoes -- Africa , Potatoes -- Therapeutic use , Medicinal plants
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:10322 , http://hdl.handle.net/10948/1228 , Potatoes -- Africa , Potatoes -- Therapeutic use , Medicinal plants
- Description: The African potato is used as an African traditional medicine for its nutritional and medicinal properties. Most research has been carried out on H. hemerocallidea, with very little or nothing on other Hypoxis spp. The main aim of this project was to provide scientific data on the anticancer, anti-inflammatory and antioxidant properties of H. hemerocallidea, H. stellipilis and H. sobolifera chloroform extracts and their active compounds. The hypoxoside and phytosterol contents of the three Hypoxis spp. were determined using TLC, HPLC and GC. H. hemerocallidea and H. sobolifera chloroform extracts contained the highest amounts of hypoxoside and β-sitosterol, respectively. For the anticancer properties, cytotoxicity of the Hypoxis extracts and its purified compounds were determined against the HeLa, HT-29 and MCF-7 cancer cell lines (using MTT), and PBMCs (using CellTiter-Blue®). H. sobolifera had the best cytotoxicity against the three cancer cell lines, whereas H. stellipilis stimulated HeLa and HT-29 cancer cell growth. IC50 values of hypoxoside and rooperol were determined. DNA cell cycle arrest (using PI staining) occurred in the late G1/early S (confirmed by increased p21Waf1/Cip1 expression) and G2/M phases after 15 and 48 hrs, respectively, when treated with Hypoxis extracts and rooperol. H. sobolifera and rooperol activated caspase-3 and -7 (using fluorescently labelled antibodies) in HeLa and HT-29 cancer cells, and caspase-7 in MCF-7 cancer cells after 48 hrs. Annexin V binding to phosphatidylserines in rooperol treated U937 cells confirmed early apoptosis after 15 hrs. The TUNEL assay showed DNA fragmentation in the three cancer cell lines when treated with H. sobolifera and rooperol for 48 hrs. A shift pass the G2/M phase has led to the investigation of endoreduplication, which was confirmed by cell/nucleus size, and anti-apoptotic proteins (Akt, phospho-Akt, phospho-Bcl-2 and p21Waf1/Cip1). U937 cell differentiation to monocyte-macrophages was optimized using PMA and 1,25(OH)2D3, which was confirmed by morphological and biochemical changes. For the anti-inflammatory properties, Hypoxis extracts and rooperol significantly increased NO production in monocyte-macrophages (pre-loaded with DAF-2 DA) and phagocytosis of pHrodoTM E. coli BioParticles®. The treatments had no effect on COX-2 expression in monocyte-macrophages. The phytosterols significantly increased IL-1β and IL-6 secretion xv (using the FlowCytomix Multiplex human Th1/Th2 10plex Kit I) in the PBMCs of one donor. For the antioxidant properties, Hypoxis extracts and rooperol significantly increased ROS production in undifferentiated and differentiated U937 cells, which were pre-loaded with DCFH-DA. Hypoxis extracts and purified compounds had ferric reducing activities, but only rooperol had ferric reducing activities significantly greater than ascorbic acid. β-sitosterol, campesterol and cholesterol significantly increased SOD activity in Chang liver cells, while H. stellipilis, H. sobolifera and rooperol decreased SOD activity. Anticancer, anti-inflammatory and antioxidant properties of the Hypoxis extracts may be attributed to the β-sitosterol content, because Hypoxis chloroform extracts contained very little or no hypoxoside. Unidentified compounds, and synergistic and additive effects of the compounds may have contributed to the biological effects. This study confirms previous reports that rooperol is the active compound. Results provide scientific data on the medicinal properties of one of the most frequently used medicinal plants in South Africa.
- Full Text:
- Date Issued: 2010
In vitro cytotoxic effects of selected Nigerian medicinal plant extracts on cancer cell lines
- Authors: Baatjies, Lucinda
- Date: 2012
- Subjects: Cancer -- Treatment , Cancer cells , Medicinal plants , Plant extracts , Traditional medicine , Public health
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10316 , http://hdl.handle.net/10948/d1008191 , Cancer -- Treatment , Cancer cells , Medicinal plants , Plant extracts , Traditional medicine , Public health
- Description: Cancer is a disease that imposes a heavy burden on public health and poses a challenge to science. The World Health Organization estimates that 80 percent of people in developing countries of the world rely on traditional medicine for their primary health needs, and about 85 percent of traditional medicine involves the use of plant extracts. This is particularly true in Africa where a large percentage of the population depends upon medicinal plants for health care. Therefore, detailed screening and evaluation of bioactive substances for chemotherapeutic purposes of African plants are urgently warranted. Furthermore, this will serve to validate the efficacy and safety of African traditional medicine. The current study investigated the in vitro cytotoxic effects of 17 ethanolic extracts of the following 16 plants used in traditional anticancer medicine in Nigeria: Sapium ellipticum leaves, Sapium ellipticum stembark, Combretum paniculatum, Celosia trigyna, Pupalia lappacea, Justica extensa, Hedranthera barteri leaves, Alternanthera sessilis, Ethulia conyzoides leaves, Lannea nigritana stembark, Combretum zenkeri root, Combretum molle leaves, Adenanthera parvoniana, Lannea acida, Cyathula achyranthoides, Drymaria cordata, Cyathula prostrata, against HeLa cancer cells. Five of the most promising extracts (Sapium ellipticum leaves, Combretum paniculatum, Celosia trigyna, Drymaria cordata, Cyathula prostrata) were selected for further screening against HT29 and MCF-7 cancer cells. Of the five, the first two were investigated further based on their activities in the screening phase. The S. ellipticum leaf extract yielded IC50 values of 88.60 ± 0.03 and 93.03 ± 0.03 μg/ml against HeLa and MCF-7, respectively. The toxicity was also evaluated on normal cells and an IC50 of 77.66 μg/ml was obtained for peripheral blood mononuclear cells (PBMCs). The IC50 values for proliferating and confluent Chang liver cells were both >125 μg/ml. These results suggest that the extract may be selective for specific cell types. Bio-assay guided fractionation of the S. ellipticum ethanolic extract yielded two active fractions; chloroform and ethyl acetate. Two compounds isolated from the chloroform extract were screened against the three cancer cell lines and found to be inactive. Three compounds were isolated from the ethyl acetate fraction and revealed IC50 values < 62.5 and < 31 μg/ml against MCF-7. Unfortunately these two compounds soon lost activity before any further work could be done on them and work was continued with the crude extract.
- Full Text:
- Date Issued: 2012
- Authors: Baatjies, Lucinda
- Date: 2012
- Subjects: Cancer -- Treatment , Cancer cells , Medicinal plants , Plant extracts , Traditional medicine , Public health
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10316 , http://hdl.handle.net/10948/d1008191 , Cancer -- Treatment , Cancer cells , Medicinal plants , Plant extracts , Traditional medicine , Public health
- Description: Cancer is a disease that imposes a heavy burden on public health and poses a challenge to science. The World Health Organization estimates that 80 percent of people in developing countries of the world rely on traditional medicine for their primary health needs, and about 85 percent of traditional medicine involves the use of plant extracts. This is particularly true in Africa where a large percentage of the population depends upon medicinal plants for health care. Therefore, detailed screening and evaluation of bioactive substances for chemotherapeutic purposes of African plants are urgently warranted. Furthermore, this will serve to validate the efficacy and safety of African traditional medicine. The current study investigated the in vitro cytotoxic effects of 17 ethanolic extracts of the following 16 plants used in traditional anticancer medicine in Nigeria: Sapium ellipticum leaves, Sapium ellipticum stembark, Combretum paniculatum, Celosia trigyna, Pupalia lappacea, Justica extensa, Hedranthera barteri leaves, Alternanthera sessilis, Ethulia conyzoides leaves, Lannea nigritana stembark, Combretum zenkeri root, Combretum molle leaves, Adenanthera parvoniana, Lannea acida, Cyathula achyranthoides, Drymaria cordata, Cyathula prostrata, against HeLa cancer cells. Five of the most promising extracts (Sapium ellipticum leaves, Combretum paniculatum, Celosia trigyna, Drymaria cordata, Cyathula prostrata) were selected for further screening against HT29 and MCF-7 cancer cells. Of the five, the first two were investigated further based on their activities in the screening phase. The S. ellipticum leaf extract yielded IC50 values of 88.60 ± 0.03 and 93.03 ± 0.03 μg/ml against HeLa and MCF-7, respectively. The toxicity was also evaluated on normal cells and an IC50 of 77.66 μg/ml was obtained for peripheral blood mononuclear cells (PBMCs). The IC50 values for proliferating and confluent Chang liver cells were both >125 μg/ml. These results suggest that the extract may be selective for specific cell types. Bio-assay guided fractionation of the S. ellipticum ethanolic extract yielded two active fractions; chloroform and ethyl acetate. Two compounds isolated from the chloroform extract were screened against the three cancer cell lines and found to be inactive. Three compounds were isolated from the ethyl acetate fraction and revealed IC50 values < 62.5 and < 31 μg/ml against MCF-7. Unfortunately these two compounds soon lost activity before any further work could be done on them and work was continued with the crude extract.
- Full Text:
- Date Issued: 2012
Phytochemical analysis and antibacterial properties of aqueous and ethanol extracts of Brachylaena elliptica (Thurb.) dc. and Brachylaena ilicifolia (Lam.) Phill & Schweick
- Authors: Sagbo, Idowu Jonas
- Date: 2015
- Subjects: Medicinal plants , Traditional medicine , Herbs -- Therapeutic use
- Language: English
- Type: Thesis , Masters , MSc (Biochemistry)
- Identifier: vital:11297 , http://hdl.handle.net/10353/d1021289 , Medicinal plants , Traditional medicine , Herbs -- Therapeutic use
- Description: Resistance of human pathogenic bacterial strains results in selective pressure against known antibiotic. However, plant derived compounds that possess antibacterial potential are currently being investigated for treatment of wound infections in diabetic patients as they are inexpensive and non-toxic. Hence, this dissertation was designed to evaluate two medicinal plants (Brachylaena elliptica and Brachylaena ilicifolia) traditionally used in the treatment of various diseases such as diabetes, and its secondary complications in diabetic patients. The in vitro antioxidant activity of both plants were evaluated using DPPH (1, 1-diphenylhydrazl), ferric reducing power, ABTS (2, 2’-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid), NO (nitric oxide) and H2O2 (hydrogen peroxide) techniques. The antibacterial test and Minimum inhibitory concentration (MIC) was determined by agar dilution method against 5 bacteria strains (Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus pyogene, Proteus vulgaris and Proteus mirabilis) infecting wounds in diabetic patients using amoxicillin and ciprofloxacin as positive control. The phytochemical analyses were assessed using standard published methods. Identification of bioactive components in essential oils of both plants were assessed using GCMS. The aqueous and ethanol extracts of both plants were also evaluated to identify bioactive components using LC-MS. The results of the phytochemical analysis revealed the presence of phenols, tannins, flavanoids, flavanols, proanthocyanidins, saponins and alkaloids in both plants. Both plants indicated strong antioxidant activities which might be due to the presence of bioactive compounds. The aqueous and ethanol leaf extracts of both plants demonstrated appreciable broad spectrum activities against these wound pathogens with MIC ranging between 5 and 0.3 mg/ml. The GC-MS analysis of the essential oils of both plants revealed the presence of monoterpenes, oxygenated sesquiterpenes, phenolics and esters. The LC-MS analysis of the aqueous and ethanol leaf extracts of both plants showed that both plants are rich in alkaloids, terpenes, terpenoids, monoterpernoids, and flavanoids. Conclusively, this study has partially justified the ethnomedicinal use of B. elliptica and B.licifolia leaves for the treatment of various diseases, including diabetes and wound infections caused by bacteria in diabetic patients. These may be attributed to the presence of antioxidant compound such as phenols, flavanoids, saponins, tannins, alkaloids and other phytochemical compounds.
- Full Text:
- Date Issued: 2015
- Authors: Sagbo, Idowu Jonas
- Date: 2015
- Subjects: Medicinal plants , Traditional medicine , Herbs -- Therapeutic use
- Language: English
- Type: Thesis , Masters , MSc (Biochemistry)
- Identifier: vital:11297 , http://hdl.handle.net/10353/d1021289 , Medicinal plants , Traditional medicine , Herbs -- Therapeutic use
- Description: Resistance of human pathogenic bacterial strains results in selective pressure against known antibiotic. However, plant derived compounds that possess antibacterial potential are currently being investigated for treatment of wound infections in diabetic patients as they are inexpensive and non-toxic. Hence, this dissertation was designed to evaluate two medicinal plants (Brachylaena elliptica and Brachylaena ilicifolia) traditionally used in the treatment of various diseases such as diabetes, and its secondary complications in diabetic patients. The in vitro antioxidant activity of both plants were evaluated using DPPH (1, 1-diphenylhydrazl), ferric reducing power, ABTS (2, 2’-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid), NO (nitric oxide) and H2O2 (hydrogen peroxide) techniques. The antibacterial test and Minimum inhibitory concentration (MIC) was determined by agar dilution method against 5 bacteria strains (Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus pyogene, Proteus vulgaris and Proteus mirabilis) infecting wounds in diabetic patients using amoxicillin and ciprofloxacin as positive control. The phytochemical analyses were assessed using standard published methods. Identification of bioactive components in essential oils of both plants were assessed using GCMS. The aqueous and ethanol extracts of both plants were also evaluated to identify bioactive components using LC-MS. The results of the phytochemical analysis revealed the presence of phenols, tannins, flavanoids, flavanols, proanthocyanidins, saponins and alkaloids in both plants. Both plants indicated strong antioxidant activities which might be due to the presence of bioactive compounds. The aqueous and ethanol leaf extracts of both plants demonstrated appreciable broad spectrum activities against these wound pathogens with MIC ranging between 5 and 0.3 mg/ml. The GC-MS analysis of the essential oils of both plants revealed the presence of monoterpenes, oxygenated sesquiterpenes, phenolics and esters. The LC-MS analysis of the aqueous and ethanol leaf extracts of both plants showed that both plants are rich in alkaloids, terpenes, terpenoids, monoterpernoids, and flavanoids. Conclusively, this study has partially justified the ethnomedicinal use of B. elliptica and B.licifolia leaves for the treatment of various diseases, including diabetes and wound infections caused by bacteria in diabetic patients. These may be attributed to the presence of antioxidant compound such as phenols, flavanoids, saponins, tannins, alkaloids and other phytochemical compounds.
- Full Text:
- Date Issued: 2015
Chang liver cell line as a model for Type II Diabetes in the liver and possible reversal of this condition by an indigenous medicinal plant
- Authors: Williams, Saralene Iona
- Date: 2009
- Subjects: Diabetes -- Alternative treatment , Medicinal plants , Traditional medicine , Liver -- Diseases
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:10339 , http://hdl.handle.net/10948/d1016179
- Description: The incidence of Type 2 Diabetes Mellittus (T2DM) is increasing world wide. In Africa the limited access to health care and the insidious course of the disease lead to more severe illness and diabetic complications. There is a need to find alternative approaches to treatment and prevention that address the problems and needs of Africa. Sutherlandia frutescens (S.frutescens) is a traditional herbal plant with known anti-diabetic properties, the precise mechanism of action of S.frutescens is not known. In order to develop new approaches for treatment and prevention of T2DM the pathophysiology of T2DM must be understood. T2DM is the final outcome of a multi-organ disease characterized by early defects in muscle, adipocytes, hepatocytes and pancreatic β-cells. In this study the role of the liver was investigated because of its central role in glucose and lipid metabolism. It is hard to differentiate between all the influences in an in vivo model, so the aim of this study was to develop an in vitro model of T2DM in Chang liver cells and to determine if S.frutescens can reverse the state of insulin resistance in this model. Different culture media conditions were screened to identify a method that can be used as the T2DM model in Chang liver cells. Serum free medium (MCBD-201) supplemented with human diabetic serum, (2.5%-10%), high insulin concentrations (0.1μM-1μM), high fructose concentrations (1-10mM). and a combination of high insulin and high fructose was used for this screening. Chang liver cells cultured in MCBD-201 medium supplemented with 1mM fructose and 0.1μM insulin showed reduced glucose uptake and increased lipid accumulation. The effect of two S.frutescens extracts, two anti-diabetic drugs, metformin and ciglitazone, and a hypolipidemic drug ciprofibrate were determined and shown to increase glucose uptake and reduce lipid accumulation. It was postulated that exposing the cells to excess nutrients in the form of high fructose would stimulate the cells to become adipogenic and accumulate lipids, which would interfere with the glucose uptake and induce insulin resistance. Gene expression of PPARγ, PPARα, and SREBP-1 transcription factors regulating lipid metabolism was determined in Chang liver cells cultured in insulin resistance inducing medium over a 48 hour time course. The expression of PPARγ, known to stimulate adipogenesis was increased after 6, 24 and 48 hours of exposure (P(H1)<0.0001). The expression of PPARα, known to stimulate β-oxidation expression, was significantly decreased after 24 hours of exposure (P(H1)<0.0001). The presence of the plant extracts in the insulin resistance inducing media protect against this increase in adipogenesis and decrease in β-oxidation after 48 hours of exposure by increasing PPARα expression and decreasing PPARγ expression. A PCR Array was performed which identified 32 more potential molecular targets of S.frutescens. Five of the 32 targets identified with the PCR Array were validated using qRT-PCR. These genes play a role in lipid and glucose metabolism and protection against oxidative stress and inflammation. In summary a cellular model of insulin resistace in hepatocytes has been established and the capacity of S.frutescens to reverse this process has been demonstrated by acting as a dual PPARγ/α agonist. New genes have been identified in the development of insulin resistance and as targets of S.frutescens.
- Full Text:
- Date Issued: 2009
- Authors: Williams, Saralene Iona
- Date: 2009
- Subjects: Diabetes -- Alternative treatment , Medicinal plants , Traditional medicine , Liver -- Diseases
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:10339 , http://hdl.handle.net/10948/d1016179
- Description: The incidence of Type 2 Diabetes Mellittus (T2DM) is increasing world wide. In Africa the limited access to health care and the insidious course of the disease lead to more severe illness and diabetic complications. There is a need to find alternative approaches to treatment and prevention that address the problems and needs of Africa. Sutherlandia frutescens (S.frutescens) is a traditional herbal plant with known anti-diabetic properties, the precise mechanism of action of S.frutescens is not known. In order to develop new approaches for treatment and prevention of T2DM the pathophysiology of T2DM must be understood. T2DM is the final outcome of a multi-organ disease characterized by early defects in muscle, adipocytes, hepatocytes and pancreatic β-cells. In this study the role of the liver was investigated because of its central role in glucose and lipid metabolism. It is hard to differentiate between all the influences in an in vivo model, so the aim of this study was to develop an in vitro model of T2DM in Chang liver cells and to determine if S.frutescens can reverse the state of insulin resistance in this model. Different culture media conditions were screened to identify a method that can be used as the T2DM model in Chang liver cells. Serum free medium (MCBD-201) supplemented with human diabetic serum, (2.5%-10%), high insulin concentrations (0.1μM-1μM), high fructose concentrations (1-10mM). and a combination of high insulin and high fructose was used for this screening. Chang liver cells cultured in MCBD-201 medium supplemented with 1mM fructose and 0.1μM insulin showed reduced glucose uptake and increased lipid accumulation. The effect of two S.frutescens extracts, two anti-diabetic drugs, metformin and ciglitazone, and a hypolipidemic drug ciprofibrate were determined and shown to increase glucose uptake and reduce lipid accumulation. It was postulated that exposing the cells to excess nutrients in the form of high fructose would stimulate the cells to become adipogenic and accumulate lipids, which would interfere with the glucose uptake and induce insulin resistance. Gene expression of PPARγ, PPARα, and SREBP-1 transcription factors regulating lipid metabolism was determined in Chang liver cells cultured in insulin resistance inducing medium over a 48 hour time course. The expression of PPARγ, known to stimulate adipogenesis was increased after 6, 24 and 48 hours of exposure (P(H1)<0.0001). The expression of PPARα, known to stimulate β-oxidation expression, was significantly decreased after 24 hours of exposure (P(H1)<0.0001). The presence of the plant extracts in the insulin resistance inducing media protect against this increase in adipogenesis and decrease in β-oxidation after 48 hours of exposure by increasing PPARα expression and decreasing PPARγ expression. A PCR Array was performed which identified 32 more potential molecular targets of S.frutescens. Five of the 32 targets identified with the PCR Array were validated using qRT-PCR. These genes play a role in lipid and glucose metabolism and protection against oxidative stress and inflammation. In summary a cellular model of insulin resistace in hepatocytes has been established and the capacity of S.frutescens to reverse this process has been demonstrated by acting as a dual PPARγ/α agonist. New genes have been identified in the development of insulin resistance and as targets of S.frutescens.
- Full Text:
- Date Issued: 2009
Pharmaceutical analysis and drug interaction studies : African potato (Hypoxis hemerocallidea)
- Purushothaman Nair, Vipin Devi Prasad
- Authors: Purushothaman Nair, Vipin Devi Prasad
- Date: 2006
- Subjects: Potatoes -- Africa , Potatoes -- Therapeutic use , AIDS (Disease) -- Treatment , HIV infections -- Drug therapy , Medicinal plants
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:3865 , http://hdl.handle.net/10962/d1015802
- Description: In order for a medicinal product to produce a consistent and reliable therapeutic response, it is essential that the final composition of the product is invariable and that the active ingredient/s is/are present in appropriate, non-toxic amounts. However, due to the complexity involved in the standardization of natural products, quality control (QC) criteria and procedures for the registration and market approval of such products are conspicuously absent in most countries around the world. African Potato (AP) is of great medical interest and this particular plant has gained tremendous popularity following the endorsement by the South African Minister of Health as a remedy for HIV/ AIDS patients. Very little information has appeared in the literature to describe methods for the quantitative analysis of hypoxoside, an important component in AP. It has also been claimed that sterols and sterolins present in AP are responsible for its medicinal property but is yet to be proven scientifically. To-date, no QC methods have been reported for the simultaneous quantitative analysis of the combination, β- sitosterol (BSS)/ stigmasterol (STG)/ stigmastanol (STN), purported to be present in preparations containing AP. The effect of concomitant administration of AP and other herbal medicines on the safety and efficacy of conventional medicines has not yet been fully determined. Amongst the objectives of this study was to develop and validate quantitative analytical methods that are suitable for the assay and quality control of plant material, extracts and commercial formulations containing AP. Hypoxoside was isolated from AP and characterized for use as a reference standard for the quality control of AP products and a stability-indicating HPLC/ UV assay method for the quantitative determination of hypoxoside was developed. In addition, a quantitative capillary zone electrophoretic (CZE) method was developed to determine hypoxoside, specifically for its advantages over HPLC. A HPLC method was also developed and validated for the quantitative analysis of BSS, STG and STN in commercially available oral dosage forms containing AP material or extracts thereof. The antioxidant activity of an aqueous extract of lyophilized corms of AP along with hypoxoside and rooperol were investigated. In comparison with the AP extracts and also with hypoxoside, rooperol showed significant antioxidant activity. The capacity of AP, (extracts, formulations, hypoxoside and rooperol as well as sterols to inhibit in vitro metabolism of drug substrates by human cytochrome P450 (CYP) enzymes such as CYP 3A4, 3A5 and CYP19 were investigated. Samples were also assessed for their effect on drug transport proteins such as P-glycoprotein (P-gp). Various extracts of AP, AP formulations, stigmasterol and the norlignans, in particular the aglycone rooperol, exhibited inhibitory effects on CYP 3A4, 3A5 and CYP19 mediated metabolism.These results suggest that concurrent therapy with AP and other medicines, in particular antiretroviral drugs, can have important implications for safety and efficacy. Large discrepancies in marker content between AP products were found. Dissolution testing of AP products was investigated as a QC tool and the results also revealed inconsistencies between different AP products.
- Full Text:
- Date Issued: 2006
- Authors: Purushothaman Nair, Vipin Devi Prasad
- Date: 2006
- Subjects: Potatoes -- Africa , Potatoes -- Therapeutic use , AIDS (Disease) -- Treatment , HIV infections -- Drug therapy , Medicinal plants
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:3865 , http://hdl.handle.net/10962/d1015802
- Description: In order for a medicinal product to produce a consistent and reliable therapeutic response, it is essential that the final composition of the product is invariable and that the active ingredient/s is/are present in appropriate, non-toxic amounts. However, due to the complexity involved in the standardization of natural products, quality control (QC) criteria and procedures for the registration and market approval of such products are conspicuously absent in most countries around the world. African Potato (AP) is of great medical interest and this particular plant has gained tremendous popularity following the endorsement by the South African Minister of Health as a remedy for HIV/ AIDS patients. Very little information has appeared in the literature to describe methods for the quantitative analysis of hypoxoside, an important component in AP. It has also been claimed that sterols and sterolins present in AP are responsible for its medicinal property but is yet to be proven scientifically. To-date, no QC methods have been reported for the simultaneous quantitative analysis of the combination, β- sitosterol (BSS)/ stigmasterol (STG)/ stigmastanol (STN), purported to be present in preparations containing AP. The effect of concomitant administration of AP and other herbal medicines on the safety and efficacy of conventional medicines has not yet been fully determined. Amongst the objectives of this study was to develop and validate quantitative analytical methods that are suitable for the assay and quality control of plant material, extracts and commercial formulations containing AP. Hypoxoside was isolated from AP and characterized for use as a reference standard for the quality control of AP products and a stability-indicating HPLC/ UV assay method for the quantitative determination of hypoxoside was developed. In addition, a quantitative capillary zone electrophoretic (CZE) method was developed to determine hypoxoside, specifically for its advantages over HPLC. A HPLC method was also developed and validated for the quantitative analysis of BSS, STG and STN in commercially available oral dosage forms containing AP material or extracts thereof. The antioxidant activity of an aqueous extract of lyophilized corms of AP along with hypoxoside and rooperol were investigated. In comparison with the AP extracts and also with hypoxoside, rooperol showed significant antioxidant activity. The capacity of AP, (extracts, formulations, hypoxoside and rooperol as well as sterols to inhibit in vitro metabolism of drug substrates by human cytochrome P450 (CYP) enzymes such as CYP 3A4, 3A5 and CYP19 were investigated. Samples were also assessed for their effect on drug transport proteins such as P-glycoprotein (P-gp). Various extracts of AP, AP formulations, stigmasterol and the norlignans, in particular the aglycone rooperol, exhibited inhibitory effects on CYP 3A4, 3A5 and CYP19 mediated metabolism.These results suggest that concurrent therapy with AP and other medicines, in particular antiretroviral drugs, can have important implications for safety and efficacy. Large discrepancies in marker content between AP products were found. Dissolution testing of AP products was investigated as a QC tool and the results also revealed inconsistencies between different AP products.
- Full Text:
- Date Issued: 2006
Phytochemical analyses and Brine shrimp (Artemia Salina) lethality studies on Syzygium cordatum
- Authors: Chiguvare, Herbert
- Date: 2013
- Subjects: Artemia , Crustacea , Chromatographic analysis , Medicinal plants , Essences and essential oils , Traditional medicine
- Language: English
- Type: Thesis , Masters , MSc (Chemistry)
- Identifier: vital:11336 , http://hdl.handle.net/10353/d1004352 , Artemia , Crustacea , Chromatographic analysis , Medicinal plants , Essences and essential oils , Traditional medicine
- Description: Syzygium cordatum Hoscht ex. C Krauss, also known as water berry, is normally used by the people of South Africa for respiratory ailments including tuberculosis, stomach complaints, treatment of wounds and as emetics. An extract of the leaves can be used as a purgative for diarrhoea treatment. The leaves of Syzygium cordatum Myrtaceae were obtained from the Eastern Cape Province of South Africa, air dried and sequential solvent extraction was done to obtain various non volatile crude extracts. The volatile extract, that is the essential oil was extracted from the leaves using hydrodistillation and analysis of compounds was done by GC/MS for composition. 32 compounds were obtained from the fresh leaves and 18 compounds were obtained from the dry leaves. The fresh oil contains caryophyllene (11.8 percent) and caryophyllene oxide (11.1 percent) as the main sesquiterpene component. α-Pinene(5.0 percent) was the only monoterpene compound identified in the fresh oil in substantial amount. The dry leaves oil had copanene (17.0 percent), β-Caryophellene (26.0 percent), cubenol (6.5 percent) and caryophellene oxide (14.2 percent) as the dominant constituent of the oil. Summary of the classes of compounds in the oil revealed that the chemical profile of both oils were dominated by sesquiterpenoid compounds. This is the first time that terpenoids compounds are being identified in both the fresh and dry leaf oil of S. cordatum. Hexane leaf extract was selected due to the interest in the terpenoid compounds. Column chromatography of the hexane crude gave five (5) of which two are fully reported. The isolates were fully elucidated using spectroscopic methods to be β-Sitosterol (HC3) and Friedela-3-one (HC1A/HC1D). Cytotoxicity analysis was carried out on the crude using the Brine shrimps assay. Isolates 1C and1D showed significant lethality using the brine shrimps assay with lethality values (LC50) of 4.105mg/ml for HC1C and 4.11mg/ml for 1D/1A respectively.
- Full Text:
- Date Issued: 2013
- Authors: Chiguvare, Herbert
- Date: 2013
- Subjects: Artemia , Crustacea , Chromatographic analysis , Medicinal plants , Essences and essential oils , Traditional medicine
- Language: English
- Type: Thesis , Masters , MSc (Chemistry)
- Identifier: vital:11336 , http://hdl.handle.net/10353/d1004352 , Artemia , Crustacea , Chromatographic analysis , Medicinal plants , Essences and essential oils , Traditional medicine
- Description: Syzygium cordatum Hoscht ex. C Krauss, also known as water berry, is normally used by the people of South Africa for respiratory ailments including tuberculosis, stomach complaints, treatment of wounds and as emetics. An extract of the leaves can be used as a purgative for diarrhoea treatment. The leaves of Syzygium cordatum Myrtaceae were obtained from the Eastern Cape Province of South Africa, air dried and sequential solvent extraction was done to obtain various non volatile crude extracts. The volatile extract, that is the essential oil was extracted from the leaves using hydrodistillation and analysis of compounds was done by GC/MS for composition. 32 compounds were obtained from the fresh leaves and 18 compounds were obtained from the dry leaves. The fresh oil contains caryophyllene (11.8 percent) and caryophyllene oxide (11.1 percent) as the main sesquiterpene component. α-Pinene(5.0 percent) was the only monoterpene compound identified in the fresh oil in substantial amount. The dry leaves oil had copanene (17.0 percent), β-Caryophellene (26.0 percent), cubenol (6.5 percent) and caryophellene oxide (14.2 percent) as the dominant constituent of the oil. Summary of the classes of compounds in the oil revealed that the chemical profile of both oils were dominated by sesquiterpenoid compounds. This is the first time that terpenoids compounds are being identified in both the fresh and dry leaf oil of S. cordatum. Hexane leaf extract was selected due to the interest in the terpenoid compounds. Column chromatography of the hexane crude gave five (5) of which two are fully reported. The isolates were fully elucidated using spectroscopic methods to be β-Sitosterol (HC3) and Friedela-3-one (HC1A/HC1D). Cytotoxicity analysis was carried out on the crude using the Brine shrimps assay. Isolates 1C and1D showed significant lethality using the brine shrimps assay with lethality values (LC50) of 4.105mg/ml for HC1C and 4.11mg/ml for 1D/1A respectively.
- Full Text:
- Date Issued: 2013
The antifungal activity of an aqueous Tulbaghia violacea plant extract against Aspergillus flavus
- Authors: Belewa, Xoliswa Vuyokazi
- Date: 2015
- Subjects: Medicinal plants , Antifungal agents , Fungi -- Biotechnology
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10948/5858 , vital:21001
- Description: Phytochemical analysis of both HEA1 and the crude plant extract showed the presence of phenolics, tannins and saponins. Saponins were the predominant secondary metabolites and were mostly abundant in the plant extract and to a lesser extent in the active compound. Steroidal saponins, tannins and phenolics were also detected in the plant extract, but only the phenolics were detected in the active compound. The results of the phytochemical analysis showed that those compounds that were not present in the active compound could be removed from the crude extract during the TLC purification process. Investigation on the mechanism of action of the crude plant extract on the sterol production by A. flavus showed that the plant extract affected ergosterol biosynthesis by causing an accumulation of oxidosqualene in the ergosterol biosynthetic pathway resulting in a decline in ergosterol production. An oscillatory response in lanosterol production was observed in the presence of the plant extract, which may be an adaptation mechanism of A. flavus to unfavourable conditions and compensation for the loss of enzyme activity which may have occurred as a result of the accumulation of oxidosqualene. The antifungal activity of the plant extract on ergosterol production by A. flavus may also be due to saponins which target the cell membrane and ergosterol production in fungi. The effect of the plant extract on the fungal cell wall of A. flavus also showed that the plant extract caused a decline in β-(1, 3) glucan production by inhibiting β-glucan synthase. The plant extract also affected the chitin synthesis pathway of A. flavus, by causing a decline in chitin production, which was due to the inhibition of chitin synthase. Investigation of chitinase production using 4MU substrates showed that the plant extract caused an accumulation of chitobioses, by activating chitobiosidases and endochitinases. A decline in N-acetylglucosaminidase activity in the presence of the plant extract was observed and this prevented the formation of N-acetylglucosamine. The accumulation of chitobiosidase and endochitinase may be as a result of autolysis that may be triggered by A. flavus as a survival mechanism in the presence of the plant extract and as a compensatory mechanism for the loss of β-glucans and chitin. The antifungal effect of the plant extract on various components of the cell wall of A. flavus, makes T. violacea aqueous plant extract an ideal chemotherapeutic agent against both human and plant pathogens of Aspergillus. The broad spectrum of antifungal activity of T. violacea against A. flavus also eliminates any chances of the fungus developing resistance towards it and would make it a candidate for use as a potential antifungal agent. Further identification and possible chemical synthesis is needed to shed light on the safety and efficacy of the active compound for further development as a chemotherapeutic agent.
- Full Text:
- Date Issued: 2015
- Authors: Belewa, Xoliswa Vuyokazi
- Date: 2015
- Subjects: Medicinal plants , Antifungal agents , Fungi -- Biotechnology
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10948/5858 , vital:21001
- Description: Phytochemical analysis of both HEA1 and the crude plant extract showed the presence of phenolics, tannins and saponins. Saponins were the predominant secondary metabolites and were mostly abundant in the plant extract and to a lesser extent in the active compound. Steroidal saponins, tannins and phenolics were also detected in the plant extract, but only the phenolics were detected in the active compound. The results of the phytochemical analysis showed that those compounds that were not present in the active compound could be removed from the crude extract during the TLC purification process. Investigation on the mechanism of action of the crude plant extract on the sterol production by A. flavus showed that the plant extract affected ergosterol biosynthesis by causing an accumulation of oxidosqualene in the ergosterol biosynthetic pathway resulting in a decline in ergosterol production. An oscillatory response in lanosterol production was observed in the presence of the plant extract, which may be an adaptation mechanism of A. flavus to unfavourable conditions and compensation for the loss of enzyme activity which may have occurred as a result of the accumulation of oxidosqualene. The antifungal activity of the plant extract on ergosterol production by A. flavus may also be due to saponins which target the cell membrane and ergosterol production in fungi. The effect of the plant extract on the fungal cell wall of A. flavus also showed that the plant extract caused a decline in β-(1, 3) glucan production by inhibiting β-glucan synthase. The plant extract also affected the chitin synthesis pathway of A. flavus, by causing a decline in chitin production, which was due to the inhibition of chitin synthase. Investigation of chitinase production using 4MU substrates showed that the plant extract caused an accumulation of chitobioses, by activating chitobiosidases and endochitinases. A decline in N-acetylglucosaminidase activity in the presence of the plant extract was observed and this prevented the formation of N-acetylglucosamine. The accumulation of chitobiosidase and endochitinase may be as a result of autolysis that may be triggered by A. flavus as a survival mechanism in the presence of the plant extract and as a compensatory mechanism for the loss of β-glucans and chitin. The antifungal effect of the plant extract on various components of the cell wall of A. flavus, makes T. violacea aqueous plant extract an ideal chemotherapeutic agent against both human and plant pathogens of Aspergillus. The broad spectrum of antifungal activity of T. violacea against A. flavus also eliminates any chances of the fungus developing resistance towards it and would make it a candidate for use as a potential antifungal agent. Further identification and possible chemical synthesis is needed to shed light on the safety and efficacy of the active compound for further development as a chemotherapeutic agent.
- Full Text:
- Date Issued: 2015
Biological activities and mechanisms of action of two ethnobotanically selected South African medicinal plants on some bacteria associated with gastrointestinal infections
- Olajuyigbe, Olufunmiso Olusola https://orcid.org/0000-0002-7889-0416
- Authors: Olajuyigbe, Olufunmiso Olusola https://orcid.org/0000-0002-7889-0416
- Date: 2012-08
- Subjects: Medicinal plants , Herbs -- Therapeutic use , Gastrointestinal system
- Language: English
- Type: Doctoral theses , text
- Identifier: http://hdl.handle.net/10353/25439 , vital:64249
- Description: In this study, 36 plant species representing 24 families were found to be commonly used for the treatment of a variety of gastrointestinal disorders in Eastern Cape, South Africa. The family Fabaceae had the highest number of species. Out of these, 47.06percent were used in the treatment of dysentery alone while 46.15percent were used in the treatment of diarrhoea. Acacia mearnsii De Wild and Ziziphus mucronata subsp. mucronata Willd were selected for this research because they are extensively used in folkloric medicine in South Africa and there was lack of scientific reports that documented their biological activities. The phytochemical screening, antioxidant activities, in vitro antimicrobial activities, cytotoxicity, the synergistic potentials and mechanisms of actions of these plants were investigated. The phytochemical screening and the antioxidant activities of the two species showed that the quantity of the phenolic compounds, flavonoids and proanthocyanidins detected differ significantly in the various extracts. Of the aqueous, acetone, ethanolic and methanolic extracts of A. mearnsii, the ethanolic extract had the highest flavonoids while the acetone extract had the highest phenolic contents. The proanthocyanidins were highest in the methanol extract while aqueous extracts had the least phytochemicals. Aqueous extract showed the least ferric reducing power but methanol extract indicated the highest reducing power. The reducing power of the extracts was lower than those obtained from the reference standard such as butylated hydroxytoluene (BHT), rutin and ascorbic acid. 2,2’-azinobis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) diammonium salt showed that ethanol extract exhibited the highest antioxidant activity at the highest concentration tested. Also, 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay indicated that ethanol extract had the highest radical scavenging activity at the lowest concentration and the activities of all the extracts decreased with increase in their concentrations. In Z. mucronata subsp. mucronata, the phenolics were significantly higher than the flavonoids and proanthocyanidin contents in all the extracts investigated. The ethanol extract had the highest antioxidant activity, followed by the acetone extract while the aqueous extract was the least active. Reacting with ABTS, the 50percent inhibitory concentrations (IC50) were (0.0429 ± 0.04 mg/ml) for aqueous, (0.0317 ± 0.04 mg/ml) for acetone and (0.0306 ± 0.04 mg/ml) for ethanol extracts while they inhibited DPPH radical with 50percent inhibitory concentration (IC50) values of 0.0646 ± 0.02 mg/ml (aqueous), 0.0482 ± 0.02 mg/ml (acetone) and 0.0422 ± 0.03 mg/ml (ethanol). The investigation showed that a positive linear correlation existed between the total phenolic content and antioxidant activity of the extracts and that these plants have strong antioxidant property and free radical scavenging capability. The in vitro antibacterial activities of Acacia mearnsii and Z. mucronata subsp. mucronata showed that their minimum inhibitory concentrations ranged between 0.039 mg/ml and 1.25 mg/ml. With the exception of acetone extract of A. mearnsii having MICs greater than 1.0 mg/ml for Enterococcus faecalis ATCC 29212 and Bacillus subtilis KZN, all other isolates had MICs less than 0.7 mg/ml. In all the bacteria treated with Z. mucronata subsp. mucronata extracts, Enterobacter cloacae ATCC 13047 had MIC greater than 1 mg/ml in methanol extract, Enterococcus faecalis ATCC 29212 and Staphylococcus aureus ATCC 6538 had MICs greater than 1 mg/ml in acetone extract while all other isolates were highly susceptible to the different extracts of Z. mucronata subsp. mucronata and had MICs less than 0.7 mg/ml. While aqueous extract was as active as the alcoholic extracts in A. mearnsii, that of Z. mucronata had no effect. The ethanol extracts exhibited the highest degree of antibacterial activity in both plants. This study, also, showed that the antifungal activity of A. mearnsii ranging 0.3125 – 5.0 mg/ml was higher than those of the different extracts of Z. mucronata subsp. mucronata ranging 1.25 – 10.0 mg/ml. It is evident from the results of the brine shrimp lethality assay that the crude extracts of A. mearnsii with the LC50 equaled 112.36 µg/ml and having the highest levels of toxicity (100percent) death at 500 μg/ml was non toxic (LC50 > 100 μg/ml) while the LC50 for Z. mucronata subsp. mucronata equaled 90.27 µg/ml indicated a low level of toxicity. The effects of combining the crude extracts of these plants with eight antibiotics were investigated by means of checkerboard and agar diffusion methods. On using the methanol extract of A. mearnsii, the agar diffusion assay showed that extract-kanamycin combination had zones of inhibition ≥ 20 ± 1.0 mm in all the bacteria tested (100percent), followed by extract chloramphenicol (90percent) > extract-ciprofloxacin = extract-tetracycline (70percent) > extract amoxicillin (60percent) > extract-nalidixic acid (50percent) > extract-erythromycin (40percent) > extract metronidazole (20percent). The checkerboard showed synergistic interaction (61.25percent), additivity/indifference (23.75percent) and antagonistic (15percent) effects. I, therefore, concluded that the antibacterial potentials of the antibiotics were improved and combining natural products with antibiotic could be a potential source of resistance-modifying agents useful against multi-drug resistant bacteria. The influences of these extracts on the ultrastructures, elemental components, protein and lipid leakages of five different bacteria were determined as the possible mechanisms of action of the extracts investigated. The scanning electron microscopy indicated varied ultrastructural changes in the morphology of bacterial cells treated with the extracts. The X-ray microanalysis showed significant differences between the elemental contents of extract-treated and untreated bacteria while lipids and proteins were leaked to a great extent from the extract-treated bacterial strains in comparison with the untreated ones. The possible mechanisms of action of the extracts may include inhibition of a significant step in peptidoglycan assembly, inhibition of metabolic processes, disruption of cell wall and cell membranes resulting in the efflux of lipid and protein in all the bacteria tested. The possible mechanism of action involved in the lipid and protein leakages in the bacterial cells could be attributed to lipid peroxidation and protein oxidation owing to the antioxidant activities of the extracts that were active beyond the protective levels. I concluded that the morphological changes and the observed leakages showed rapid killing, significant membrane depolarization resulting in leakages and efflux of disintegrated cellular materials. In general, this study has justified the ethnotherapeutic importance of A. mearnsii and Z. mucronata subsp. mucronata in the treatment of microbial infections by indicating the possible mechanisms of action of the crude extracts on the tested bacteria. , Thesis (PhD) -- Faculty of Science and Agriculture, 2012
- Full Text:
- Date Issued: 2012-08
- Authors: Olajuyigbe, Olufunmiso Olusola https://orcid.org/0000-0002-7889-0416
- Date: 2012-08
- Subjects: Medicinal plants , Herbs -- Therapeutic use , Gastrointestinal system
- Language: English
- Type: Doctoral theses , text
- Identifier: http://hdl.handle.net/10353/25439 , vital:64249
- Description: In this study, 36 plant species representing 24 families were found to be commonly used for the treatment of a variety of gastrointestinal disorders in Eastern Cape, South Africa. The family Fabaceae had the highest number of species. Out of these, 47.06percent were used in the treatment of dysentery alone while 46.15percent were used in the treatment of diarrhoea. Acacia mearnsii De Wild and Ziziphus mucronata subsp. mucronata Willd were selected for this research because they are extensively used in folkloric medicine in South Africa and there was lack of scientific reports that documented their biological activities. The phytochemical screening, antioxidant activities, in vitro antimicrobial activities, cytotoxicity, the synergistic potentials and mechanisms of actions of these plants were investigated. The phytochemical screening and the antioxidant activities of the two species showed that the quantity of the phenolic compounds, flavonoids and proanthocyanidins detected differ significantly in the various extracts. Of the aqueous, acetone, ethanolic and methanolic extracts of A. mearnsii, the ethanolic extract had the highest flavonoids while the acetone extract had the highest phenolic contents. The proanthocyanidins were highest in the methanol extract while aqueous extracts had the least phytochemicals. Aqueous extract showed the least ferric reducing power but methanol extract indicated the highest reducing power. The reducing power of the extracts was lower than those obtained from the reference standard such as butylated hydroxytoluene (BHT), rutin and ascorbic acid. 2,2’-azinobis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) diammonium salt showed that ethanol extract exhibited the highest antioxidant activity at the highest concentration tested. Also, 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay indicated that ethanol extract had the highest radical scavenging activity at the lowest concentration and the activities of all the extracts decreased with increase in their concentrations. In Z. mucronata subsp. mucronata, the phenolics were significantly higher than the flavonoids and proanthocyanidin contents in all the extracts investigated. The ethanol extract had the highest antioxidant activity, followed by the acetone extract while the aqueous extract was the least active. Reacting with ABTS, the 50percent inhibitory concentrations (IC50) were (0.0429 ± 0.04 mg/ml) for aqueous, (0.0317 ± 0.04 mg/ml) for acetone and (0.0306 ± 0.04 mg/ml) for ethanol extracts while they inhibited DPPH radical with 50percent inhibitory concentration (IC50) values of 0.0646 ± 0.02 mg/ml (aqueous), 0.0482 ± 0.02 mg/ml (acetone) and 0.0422 ± 0.03 mg/ml (ethanol). The investigation showed that a positive linear correlation existed between the total phenolic content and antioxidant activity of the extracts and that these plants have strong antioxidant property and free radical scavenging capability. The in vitro antibacterial activities of Acacia mearnsii and Z. mucronata subsp. mucronata showed that their minimum inhibitory concentrations ranged between 0.039 mg/ml and 1.25 mg/ml. With the exception of acetone extract of A. mearnsii having MICs greater than 1.0 mg/ml for Enterococcus faecalis ATCC 29212 and Bacillus subtilis KZN, all other isolates had MICs less than 0.7 mg/ml. In all the bacteria treated with Z. mucronata subsp. mucronata extracts, Enterobacter cloacae ATCC 13047 had MIC greater than 1 mg/ml in methanol extract, Enterococcus faecalis ATCC 29212 and Staphylococcus aureus ATCC 6538 had MICs greater than 1 mg/ml in acetone extract while all other isolates were highly susceptible to the different extracts of Z. mucronata subsp. mucronata and had MICs less than 0.7 mg/ml. While aqueous extract was as active as the alcoholic extracts in A. mearnsii, that of Z. mucronata had no effect. The ethanol extracts exhibited the highest degree of antibacterial activity in both plants. This study, also, showed that the antifungal activity of A. mearnsii ranging 0.3125 – 5.0 mg/ml was higher than those of the different extracts of Z. mucronata subsp. mucronata ranging 1.25 – 10.0 mg/ml. It is evident from the results of the brine shrimp lethality assay that the crude extracts of A. mearnsii with the LC50 equaled 112.36 µg/ml and having the highest levels of toxicity (100percent) death at 500 μg/ml was non toxic (LC50 > 100 μg/ml) while the LC50 for Z. mucronata subsp. mucronata equaled 90.27 µg/ml indicated a low level of toxicity. The effects of combining the crude extracts of these plants with eight antibiotics were investigated by means of checkerboard and agar diffusion methods. On using the methanol extract of A. mearnsii, the agar diffusion assay showed that extract-kanamycin combination had zones of inhibition ≥ 20 ± 1.0 mm in all the bacteria tested (100percent), followed by extract chloramphenicol (90percent) > extract-ciprofloxacin = extract-tetracycline (70percent) > extract amoxicillin (60percent) > extract-nalidixic acid (50percent) > extract-erythromycin (40percent) > extract metronidazole (20percent). The checkerboard showed synergistic interaction (61.25percent), additivity/indifference (23.75percent) and antagonistic (15percent) effects. I, therefore, concluded that the antibacterial potentials of the antibiotics were improved and combining natural products with antibiotic could be a potential source of resistance-modifying agents useful against multi-drug resistant bacteria. The influences of these extracts on the ultrastructures, elemental components, protein and lipid leakages of five different bacteria were determined as the possible mechanisms of action of the extracts investigated. The scanning electron microscopy indicated varied ultrastructural changes in the morphology of bacterial cells treated with the extracts. The X-ray microanalysis showed significant differences between the elemental contents of extract-treated and untreated bacteria while lipids and proteins were leaked to a great extent from the extract-treated bacterial strains in comparison with the untreated ones. The possible mechanisms of action of the extracts may include inhibition of a significant step in peptidoglycan assembly, inhibition of metabolic processes, disruption of cell wall and cell membranes resulting in the efflux of lipid and protein in all the bacteria tested. The possible mechanism of action involved in the lipid and protein leakages in the bacterial cells could be attributed to lipid peroxidation and protein oxidation owing to the antioxidant activities of the extracts that were active beyond the protective levels. I concluded that the morphological changes and the observed leakages showed rapid killing, significant membrane depolarization resulting in leakages and efflux of disintegrated cellular materials. In general, this study has justified the ethnotherapeutic importance of A. mearnsii and Z. mucronata subsp. mucronata in the treatment of microbial infections by indicating the possible mechanisms of action of the crude extracts on the tested bacteria. , Thesis (PhD) -- Faculty of Science and Agriculture, 2012
- Full Text:
- Date Issued: 2012-08
Pharmacological, toxicological and phytochemical evaluation of helichrysum petiolare hilliard & b.l. burtt - an indigenous plant traditionally used in the treatment of diabetes in the eastern cape province of South Africa
- Aladejana, Adebowale Emmanuel https://orcid.org/0000-0003-1871-926X
- Authors: Aladejana, Adebowale Emmanuel https://orcid.org/0000-0003-1871-926X
- Date: 2022-04
- Subjects: Diabetes -- Alternative treatment , Traditional medicine , Medicinal plants
- Language: English
- Type: Doctoral theses , text
- Identifier: http://hdl.handle.net/10353/22787 , vital:52755
- Description: Diabetes mellitus is one of the leading causes of death in South Africa, and it has already placed significant stress on the country’s health sector and economy. The orthodox hypoglycaemic drugs are not only ineffective in the management of the disease and its complications, but they also possess unwanted side effects. The need for alternative non-toxic drugs is therefore imperative. Various studies have listed several medicinal plants that can be successfully used in the herbal treatment of diabetes and have investigated them for their anti-diabetic potentials in vivo and/or in vitro. Out of the different potential herbal species, plants belonging to the Asteraceae family possess highly potent hypoglycaemic properties with negligible toxicities. Five Asteraceae plants widely used in different parts of South Africa for the treatment of diabetes were reviewed. The review provided an update of scientific evidence on the hypoglycaemic properties of the plants. However, Helichrysum petiolare was studied extensively in this study for its antidiabetic activity H. petiolare has been listed in many ethnobotanical surveys as a plant with potent hypoglycaemic potential, this, however, has not been properly verified in scientific literature and there has hardly been any study on the essential oil and nutritional composition, and antioxidant, antidiabetic, and cytotoxicity potentials of the plant. The effects of hydro-distillation (HD) and solvent-free microwave extraction (SFME) methods on the chemical constituents of H. petiolare-derived essential oils were evaluated. The SFME method had a higher yield of essential oil than the HD. There were substantial amounts of monoterpenes, monoterpene alcohols, sesquiterpenes, and sesquiterpene alcohols in both essential oils obtained, but these compounds were more profound in the SFME derived essential oil which has 62 compounds compared to the 52 derived through HD. The SFME derived essential oil can therefore be said to be of better quality than the HD method. The compounds obtained in the essential oils have high pharmaceutical and cosmetic value, and as observed in this study, their quantity is dependent on the method of extraction (Ibáñez and Blázquez, 2021; Kaur et al., 2021). The proximate analysis of the whole plant of H. petiolare showed high levels of Acid Detergent Fibre (ADF), vitamins (A, C and E), Neutral Detergent Fibre (NDF), and minerals. The high ADF level is believed to be responsible for the low energy, fat and carbohydrate levels observed in the study. The result showed a high level of oxalate and therefore suggests cooking of the plant before human consumption. Overall nutrition and mineral compositions of the plant showed that H. petiolare is immensely rich in vital nutrients that are of great importance to health and metabolism; these nutrients are suggested to be partly responsible for the plant’s useful medicinal properties. The phytochemical contents of the acetone (ACQ), ethanol (ETQ), and boiled (BAQ) and cold (CAQ) aqueous whole-plant extracts of Helichrysum petiolare were determined using standard phytochemical reaction methods. ABTS, DPPH, NO and TAC assays were used to evaluate their antioxidant properties. The highest total phenolic content (212,963 mg/g) was reported in the BAQ extract, while the ETQ had the highest flavonoid (172.393 mg/g) and proanthocyanidin contents (65.855 mg/g). Alkaloids, flavonols, and saponin were highest in the ACQ extract, while the CAQ had the lowest phytochemical content. Among the extracts, the BAQ had the highest DPPH•+ (IC50 0.02 mg/mL) and ABTS•+ (IC50 0.07) inhibition capacities, while the ETQ exhibited the highest NO• Inhibition (IC50 0.41 mg/mL) and TAC (IC50 0.19 mg/mL). These findings justify the use of H. petiolare in traditional medicine and further recommend the ETQ and BAQ extracts of the plant as more effective extracts for medicinal treatment. The hepatotoxicity (cytotoxicity, mitotoxicity and lipotoxicity) potential of the BAQ, CAQ and ETQ extracts of Helichrysum petiolare was evaluated using standard procedures. The results showed negligible BAQ and CAQ cytotoxicities, which were further, corroborated by stability in the mitochondrial membrane potentials and were congruent with the CAQ and BAQ results for steatosis and phospholipidosis. The data suggested favourable CAQ and BAQ toxicity profiles with limited risks for hepatotoxicity. The ETQ extract, however, showed significantly high levels of cytotoxicity and lipotoxicity, and a low level of mitotoxicity. Our result suggested a potential risk of the ETQ extract for hepatotoxicity but appears partly independent of direct mitochondrial involvement. Glucose uptake assay showed significantly increased glucose uptake in the BAQ and CAQ treated L6 and C3A cell lines. The CAQ extract enhanced glucose uptake more in the L6 myocytes than in the C3A cell-lines hepatocytes. The BAQ extract showed higher levels of inhibition on α–amylase and α-glucosidase activities as compared to CAQ. The BAQ and CAQ extracts of H. petiolare may, therefore, contain pharmacologically active and relatively non-toxic hypoglycaemic chemicals, which may be effective substitutes in the treatment of diabetes mellitus. This study provides up to date scientific information on the use of H. petiolare in the treatment of diabetes mellitus in the Eastern Cape of South Africa. It justifies the use of this plant in herbal medicine and sheds more light on its previously vaguely understood nutritional and medicinal potentials. More studies, however, need to be done to isolate, identify and purify the constituent bioactive compound(s). Their dosage of application and mode of action also needs to be understood. , Thesis (PhD) -- Faculty of Science and Agriculture, 2022
- Full Text:
- Date Issued: 2022-04
- Authors: Aladejana, Adebowale Emmanuel https://orcid.org/0000-0003-1871-926X
- Date: 2022-04
- Subjects: Diabetes -- Alternative treatment , Traditional medicine , Medicinal plants
- Language: English
- Type: Doctoral theses , text
- Identifier: http://hdl.handle.net/10353/22787 , vital:52755
- Description: Diabetes mellitus is one of the leading causes of death in South Africa, and it has already placed significant stress on the country’s health sector and economy. The orthodox hypoglycaemic drugs are not only ineffective in the management of the disease and its complications, but they also possess unwanted side effects. The need for alternative non-toxic drugs is therefore imperative. Various studies have listed several medicinal plants that can be successfully used in the herbal treatment of diabetes and have investigated them for their anti-diabetic potentials in vivo and/or in vitro. Out of the different potential herbal species, plants belonging to the Asteraceae family possess highly potent hypoglycaemic properties with negligible toxicities. Five Asteraceae plants widely used in different parts of South Africa for the treatment of diabetes were reviewed. The review provided an update of scientific evidence on the hypoglycaemic properties of the plants. However, Helichrysum petiolare was studied extensively in this study for its antidiabetic activity H. petiolare has been listed in many ethnobotanical surveys as a plant with potent hypoglycaemic potential, this, however, has not been properly verified in scientific literature and there has hardly been any study on the essential oil and nutritional composition, and antioxidant, antidiabetic, and cytotoxicity potentials of the plant. The effects of hydro-distillation (HD) and solvent-free microwave extraction (SFME) methods on the chemical constituents of H. petiolare-derived essential oils were evaluated. The SFME method had a higher yield of essential oil than the HD. There were substantial amounts of monoterpenes, monoterpene alcohols, sesquiterpenes, and sesquiterpene alcohols in both essential oils obtained, but these compounds were more profound in the SFME derived essential oil which has 62 compounds compared to the 52 derived through HD. The SFME derived essential oil can therefore be said to be of better quality than the HD method. The compounds obtained in the essential oils have high pharmaceutical and cosmetic value, and as observed in this study, their quantity is dependent on the method of extraction (Ibáñez and Blázquez, 2021; Kaur et al., 2021). The proximate analysis of the whole plant of H. petiolare showed high levels of Acid Detergent Fibre (ADF), vitamins (A, C and E), Neutral Detergent Fibre (NDF), and minerals. The high ADF level is believed to be responsible for the low energy, fat and carbohydrate levels observed in the study. The result showed a high level of oxalate and therefore suggests cooking of the plant before human consumption. Overall nutrition and mineral compositions of the plant showed that H. petiolare is immensely rich in vital nutrients that are of great importance to health and metabolism; these nutrients are suggested to be partly responsible for the plant’s useful medicinal properties. The phytochemical contents of the acetone (ACQ), ethanol (ETQ), and boiled (BAQ) and cold (CAQ) aqueous whole-plant extracts of Helichrysum petiolare were determined using standard phytochemical reaction methods. ABTS, DPPH, NO and TAC assays were used to evaluate their antioxidant properties. The highest total phenolic content (212,963 mg/g) was reported in the BAQ extract, while the ETQ had the highest flavonoid (172.393 mg/g) and proanthocyanidin contents (65.855 mg/g). Alkaloids, flavonols, and saponin were highest in the ACQ extract, while the CAQ had the lowest phytochemical content. Among the extracts, the BAQ had the highest DPPH•+ (IC50 0.02 mg/mL) and ABTS•+ (IC50 0.07) inhibition capacities, while the ETQ exhibited the highest NO• Inhibition (IC50 0.41 mg/mL) and TAC (IC50 0.19 mg/mL). These findings justify the use of H. petiolare in traditional medicine and further recommend the ETQ and BAQ extracts of the plant as more effective extracts for medicinal treatment. The hepatotoxicity (cytotoxicity, mitotoxicity and lipotoxicity) potential of the BAQ, CAQ and ETQ extracts of Helichrysum petiolare was evaluated using standard procedures. The results showed negligible BAQ and CAQ cytotoxicities, which were further, corroborated by stability in the mitochondrial membrane potentials and were congruent with the CAQ and BAQ results for steatosis and phospholipidosis. The data suggested favourable CAQ and BAQ toxicity profiles with limited risks for hepatotoxicity. The ETQ extract, however, showed significantly high levels of cytotoxicity and lipotoxicity, and a low level of mitotoxicity. Our result suggested a potential risk of the ETQ extract for hepatotoxicity but appears partly independent of direct mitochondrial involvement. Glucose uptake assay showed significantly increased glucose uptake in the BAQ and CAQ treated L6 and C3A cell lines. The CAQ extract enhanced glucose uptake more in the L6 myocytes than in the C3A cell-lines hepatocytes. The BAQ extract showed higher levels of inhibition on α–amylase and α-glucosidase activities as compared to CAQ. The BAQ and CAQ extracts of H. petiolare may, therefore, contain pharmacologically active and relatively non-toxic hypoglycaemic chemicals, which may be effective substitutes in the treatment of diabetes mellitus. This study provides up to date scientific information on the use of H. petiolare in the treatment of diabetes mellitus in the Eastern Cape of South Africa. It justifies the use of this plant in herbal medicine and sheds more light on its previously vaguely understood nutritional and medicinal potentials. More studies, however, need to be done to isolate, identify and purify the constituent bioactive compound(s). Their dosage of application and mode of action also needs to be understood. , Thesis (PhD) -- Faculty of Science and Agriculture, 2022
- Full Text:
- Date Issued: 2022-04
Evaluation of the toxicity of secondary metabolites in Solanum incanum L. to advance community knowledge
- Authors: Zivanayi, William
- Date: 2023-04
- Subjects: Solanum -- Zimbabwe , Pesticides -- Toxicology , Medicinal plants
- Language: English
- Type: Doctoral theses , text
- Identifier: http://hdl.handle.net/10948/61018 , vital:69686
- Description: The effects of pests and the need to produce adequate food have influenced small-scale farmers in disadvantaged communities to adopt and utilise natural plant pesticides to improve harvests in many Southern African Development Communities. However, the phytochemistry associated with these indigenous plants’ pesticide activity still needs to be explored. The lack of evidence of scientific knowledge of the plant species has caused a lot of health issues among the users of indigenous plant pesticides. Solanum incanum is among the plants utilised to control cabbage aphids in Mkoba village, Zimbabwe. Solanum species are known for their steroidal compounds which comprise glycoalkaloids and saponins. This study evaluated the knowledge, opinions, and attitudes of the vegetable peasant farming community in Gweru regarding their use of the indigenous plant (S. incanum) as a pesticide. The study also reported the phytochemical profiling, structural characterisation of the isolated compounds, and biological and pesticidal activity evaluation of phytochemicals isolated from S. incanum. A descriptive survey was carried out to assess the knowledge, attitude, and practices of a conveniently sampled group of vegetable farmers in Mkoba village who use S. incanum as a pesticide. Forty-nine respondents comprised of 19 males and 30 females of ages ranging from 15 to above 60 years took part in the study by answering an open and closed-ended questionnaire. The survey revealed that parents and neighbours were instrumental in disseminating pesticidal information in the community. Brassica napus were the most grown.vegetable and vulnerable to cabbage aphids. Mixed opinions amongst the respondents varied regarding the health and environmental impact of S. incanum as a pesticide. Seventy-five percent (75%) of the respondents supported the use of S. incanum as a pesticide whilst 25% claimed that the use of S. incanum was the source of the health problems experienced in the community. The survey demonstrated that (45)91% of the farmers displayed poor practices regarding the disposal of empty pesticide containers and the use of personal protective clothing. The most prevalent symptoms in the community were skin rash, nausea, headache, and poor vision and these symptoms were common in the age group 30 to 60 years. , Thesis (PhD) -- Faculty of Science, School of biomolecular and Chemical Sciences, 2023
- Full Text:
- Date Issued: 2023-04
- Authors: Zivanayi, William
- Date: 2023-04
- Subjects: Solanum -- Zimbabwe , Pesticides -- Toxicology , Medicinal plants
- Language: English
- Type: Doctoral theses , text
- Identifier: http://hdl.handle.net/10948/61018 , vital:69686
- Description: The effects of pests and the need to produce adequate food have influenced small-scale farmers in disadvantaged communities to adopt and utilise natural plant pesticides to improve harvests in many Southern African Development Communities. However, the phytochemistry associated with these indigenous plants’ pesticide activity still needs to be explored. The lack of evidence of scientific knowledge of the plant species has caused a lot of health issues among the users of indigenous plant pesticides. Solanum incanum is among the plants utilised to control cabbage aphids in Mkoba village, Zimbabwe. Solanum species are known for their steroidal compounds which comprise glycoalkaloids and saponins. This study evaluated the knowledge, opinions, and attitudes of the vegetable peasant farming community in Gweru regarding their use of the indigenous plant (S. incanum) as a pesticide. The study also reported the phytochemical profiling, structural characterisation of the isolated compounds, and biological and pesticidal activity evaluation of phytochemicals isolated from S. incanum. A descriptive survey was carried out to assess the knowledge, attitude, and practices of a conveniently sampled group of vegetable farmers in Mkoba village who use S. incanum as a pesticide. Forty-nine respondents comprised of 19 males and 30 females of ages ranging from 15 to above 60 years took part in the study by answering an open and closed-ended questionnaire. The survey revealed that parents and neighbours were instrumental in disseminating pesticidal information in the community. Brassica napus were the most grown.vegetable and vulnerable to cabbage aphids. Mixed opinions amongst the respondents varied regarding the health and environmental impact of S. incanum as a pesticide. Seventy-five percent (75%) of the respondents supported the use of S. incanum as a pesticide whilst 25% claimed that the use of S. incanum was the source of the health problems experienced in the community. The survey demonstrated that (45)91% of the farmers displayed poor practices regarding the disposal of empty pesticide containers and the use of personal protective clothing. The most prevalent symptoms in the community were skin rash, nausea, headache, and poor vision and these symptoms were common in the age group 30 to 60 years. , Thesis (PhD) -- Faculty of Science, School of biomolecular and Chemical Sciences, 2023
- Full Text:
- Date Issued: 2023-04
Variation in the essential oil composition of Calendula Officinalis L
- Authors: Okoh, Omobola Oluranti
- Date: 2008
- Subjects: Calendula (Genus) , Essences and essential oils , Medicinal plants , Calendula officinalis
- Language: English
- Type: Thesis , Masters , MSc (Chemistry)
- Identifier: vital:11334 , http://hdl.handle.net/10353/d1001150 , Calendula (Genus) , Essences and essential oils , Medicinal plants , Calendula officinalis
- Description: Variations in the yield, chemical composition, antibacterial, and antioxidant properties of the essential oils of Rosmarinus officinalis L. cultivated in Alice, Eastern Cape of South Africa over a period of 12 months using the solvent-free microwave extraction and traditional hydrodistillation methods were evaluated. The GC-MS analyses of the essential oils revealed the presence of 33 compounds with 1,8-cineole, a-pinene, camphor, verbenone, bornyl acetate and camphene constituting about 80 percent of the oils throughout the period of investigation, with the solvent-free microwave extraction method generally yielding more of the major components than the hydrodistillation method. Each of the major components of the oils varied in quantity and quality of yield at different periods of the year. The method of extraction and time of harvest are of importance to the quantity and quality of essential oil of Rosmarinus officinalis. Higher amounts of oxygenated monoterpenes such as borneol, camphor, terpene- 4-ol, linalool, a-terpeneol were present in the oil of SFME in comparison with HD. However, HD oil contained more monoterpene hydrocarbons such as a-pinene, camphene, β-pinene, myrcene, a-phellanderene, 1,8-cineole, trans- β-ocimene, γ-teprinene, and cis-sabinene hydrate than SFME extracted oil. Accumulation of monoterpene alcohols and ketones was observed during maturation process of Rosmarinus leaves. Quantitative evaluation of antibacterial activity, minimum inhibitory concentration values were determined using a serial microplate dilution method. The essential oils obtained using both methods of extraction were active against all the bacteria tested at a concentration of 10 mg mL-1. The minimum inhibitory concentrations for the SFME extracted oils ranged between 0.23 and 1.88 mg mL-1, while those of the HD extracted oils varied between 0.94 and 7.5 mg mL-1, thus suggesting that the oil obtained by solvent free microwave extraction was more active against bacteria than the oil obtained through hydrodistillation. The antioxidant and free radical scavenging activity of the obtained oils were tested by means of 1,1-diphenyl-2-picrylhydrazyl radical (DPPH+) assay and β- carotene bleaching test. In the DPPH+ assay, while the free radical scavenging activity of the oil obtained by SFME method showed percentage inhibitions of between 48.8 percent and 67 percent, the HD derived oil showed inhibitions of between 52.2 percent and 65.30 percent at concentrations of 0.33, 0.50 and 1.0 mg mL-1, respectively. In the β-carotene bleaching assay, the percentage inhibition increased with increasing concentration of both oils with a higher antioxidant activity of the oil obtained through the SFME than the HD method. Thin layer chromatography (TLC) was used to analyze the chemical composition of the extracts using three eluent solvent systems of varying polarities i. e. CEF, BEA and EMW and sprayed with vanillin-sulfuric acid. The chemical composition of the different extracts was similar with the exception of methanol and water extracts which had only one or two visible compounds after treating with vanillin-spray reagent. To evaluate the number of antibacterial compounds present in the fractions, bioautography was used against two most important nosocomial microorganisms. S. aureus (Gram positive) and E. coli (Gram negative). Nearly all the crude serial extraction fractions contained compounds that inhibited the growth of E. coli. The hexane extract had the most lines of inhibition followed by ethyl acetate. Bioassay-guided fractionation against E. coli was used to isolate antibacterial compounds. The largest number of antibacterial compounds occurred in the hexane fraction. Furthermore we tried to complete the characterization by extracting and studying other biologically important plant metabolites such as phenolic compounds to evaluate the antioxidant capacity of Rosmarinus extracts
- Full Text:
- Date Issued: 2008
- Authors: Okoh, Omobola Oluranti
- Date: 2008
- Subjects: Calendula (Genus) , Essences and essential oils , Medicinal plants , Calendula officinalis
- Language: English
- Type: Thesis , Masters , MSc (Chemistry)
- Identifier: vital:11334 , http://hdl.handle.net/10353/d1001150 , Calendula (Genus) , Essences and essential oils , Medicinal plants , Calendula officinalis
- Description: Variations in the yield, chemical composition, antibacterial, and antioxidant properties of the essential oils of Rosmarinus officinalis L. cultivated in Alice, Eastern Cape of South Africa over a period of 12 months using the solvent-free microwave extraction and traditional hydrodistillation methods were evaluated. The GC-MS analyses of the essential oils revealed the presence of 33 compounds with 1,8-cineole, a-pinene, camphor, verbenone, bornyl acetate and camphene constituting about 80 percent of the oils throughout the period of investigation, with the solvent-free microwave extraction method generally yielding more of the major components than the hydrodistillation method. Each of the major components of the oils varied in quantity and quality of yield at different periods of the year. The method of extraction and time of harvest are of importance to the quantity and quality of essential oil of Rosmarinus officinalis. Higher amounts of oxygenated monoterpenes such as borneol, camphor, terpene- 4-ol, linalool, a-terpeneol were present in the oil of SFME in comparison with HD. However, HD oil contained more monoterpene hydrocarbons such as a-pinene, camphene, β-pinene, myrcene, a-phellanderene, 1,8-cineole, trans- β-ocimene, γ-teprinene, and cis-sabinene hydrate than SFME extracted oil. Accumulation of monoterpene alcohols and ketones was observed during maturation process of Rosmarinus leaves. Quantitative evaluation of antibacterial activity, minimum inhibitory concentration values were determined using a serial microplate dilution method. The essential oils obtained using both methods of extraction were active against all the bacteria tested at a concentration of 10 mg mL-1. The minimum inhibitory concentrations for the SFME extracted oils ranged between 0.23 and 1.88 mg mL-1, while those of the HD extracted oils varied between 0.94 and 7.5 mg mL-1, thus suggesting that the oil obtained by solvent free microwave extraction was more active against bacteria than the oil obtained through hydrodistillation. The antioxidant and free radical scavenging activity of the obtained oils were tested by means of 1,1-diphenyl-2-picrylhydrazyl radical (DPPH+) assay and β- carotene bleaching test. In the DPPH+ assay, while the free radical scavenging activity of the oil obtained by SFME method showed percentage inhibitions of between 48.8 percent and 67 percent, the HD derived oil showed inhibitions of between 52.2 percent and 65.30 percent at concentrations of 0.33, 0.50 and 1.0 mg mL-1, respectively. In the β-carotene bleaching assay, the percentage inhibition increased with increasing concentration of both oils with a higher antioxidant activity of the oil obtained through the SFME than the HD method. Thin layer chromatography (TLC) was used to analyze the chemical composition of the extracts using three eluent solvent systems of varying polarities i. e. CEF, BEA and EMW and sprayed with vanillin-sulfuric acid. The chemical composition of the different extracts was similar with the exception of methanol and water extracts which had only one or two visible compounds after treating with vanillin-spray reagent. To evaluate the number of antibacterial compounds present in the fractions, bioautography was used against two most important nosocomial microorganisms. S. aureus (Gram positive) and E. coli (Gram negative). Nearly all the crude serial extraction fractions contained compounds that inhibited the growth of E. coli. The hexane extract had the most lines of inhibition followed by ethyl acetate. Bioassay-guided fractionation against E. coli was used to isolate antibacterial compounds. The largest number of antibacterial compounds occurred in the hexane fraction. Furthermore we tried to complete the characterization by extracting and studying other biologically important plant metabolites such as phenolic compounds to evaluate the antioxidant capacity of Rosmarinus extracts
- Full Text:
- Date Issued: 2008