Evaluating antidiabetic properties of selected African medicinal plants in a cell-based model
- Sirkhotte, Saeedah, Reddy, Shanika
- Authors: Sirkhotte, Saeedah , Reddy, Shanika
- Date: 2023-12
- Subjects: Medicinal plants -- South Africa , Insulin resistance , Gluconeogenesis
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10948/62600 , vital:72827
- Description: The World Health Organisation lists diabetes as one of the major non-communicable diseases affecting the world, and its prevalence is expected to increase rapidly. Type II diabetes mellitus (T2DM) is characterised by insulin resistance and impaired blood glucose control. T2DM is of growing concern within South Africa, with 10.8% of the population currently diagnosed. The popularity, as well as availability, of traditional plant-based medicine in South Africa, might provide a solution. Antidiabetic potential is commonly tested by in vitro assays, however the methods to test this potential via gluconeogenesis are limited. This project aimed to optimise an in vitro hepatic gluconeogenesis model. In addition, this project aimed to determine the antidiabetic properties of Prunus africana, Hypoxis stellipilis, and Eriocephalus africanus by in vitro analysis. These properties were examined in human hepatoma cells (C3A cell line) and rat pancreatic cells (INS1 cell line) and methods included: cytotoxicity analysis, Amplex® red glucose oxidase assay, antibody staining, gene expression analysis by RT-qPCR, oxidative stress analysis, and calcium signaling for insulin release. Insulin resistance was successfully induced by exposing C3A hepatocarcinoma cells to a combination treatment of 50 μM dexamethasone, 1.25 mM fructose and 0.125 mM palmitic acid for a period of three days. Thereafter, gluconeogenesis was assessed using the Amplex® red glucose oxidase assay. The established model was effective in inducing insulin resistance and upregulating gluconeogenesis. Of the tested plant extracts, H. stellipilis showed the most potential as an antidiabetic treatment. It had low toxicity, significantly decreased hepatic glucose production and reduced the amount of phosphoenolpyruvate carboxykinase (PCK) and well as PCK gene expression, and reduced lipid content and reactive oxygen species (ROS) in the C3A cell line. H. stellipilis increased calcium signalling in INS1 rat insulinoma cells, however there was a decrease in expression of genes for insulin and glucose transporter 2 after 6 hr exposure. H. stellipilis appears be beneficial as an antidiabetic treatment. Although antidiabetic studies have been done on other Hypoxis species, this is the first study on the effects of H. stellipilis on gluconeogenesis and diabetes. , Thesis (MSc) -- Faculty of Science, School of Biomolecular & Chemical Sciences, 2023
- Full Text:
- Date Issued: 2023-12
- Authors: Sirkhotte, Saeedah , Reddy, Shanika
- Date: 2023-12
- Subjects: Medicinal plants -- South Africa , Insulin resistance , Gluconeogenesis
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10948/62600 , vital:72827
- Description: The World Health Organisation lists diabetes as one of the major non-communicable diseases affecting the world, and its prevalence is expected to increase rapidly. Type II diabetes mellitus (T2DM) is characterised by insulin resistance and impaired blood glucose control. T2DM is of growing concern within South Africa, with 10.8% of the population currently diagnosed. The popularity, as well as availability, of traditional plant-based medicine in South Africa, might provide a solution. Antidiabetic potential is commonly tested by in vitro assays, however the methods to test this potential via gluconeogenesis are limited. This project aimed to optimise an in vitro hepatic gluconeogenesis model. In addition, this project aimed to determine the antidiabetic properties of Prunus africana, Hypoxis stellipilis, and Eriocephalus africanus by in vitro analysis. These properties were examined in human hepatoma cells (C3A cell line) and rat pancreatic cells (INS1 cell line) and methods included: cytotoxicity analysis, Amplex® red glucose oxidase assay, antibody staining, gene expression analysis by RT-qPCR, oxidative stress analysis, and calcium signaling for insulin release. Insulin resistance was successfully induced by exposing C3A hepatocarcinoma cells to a combination treatment of 50 μM dexamethasone, 1.25 mM fructose and 0.125 mM palmitic acid for a period of three days. Thereafter, gluconeogenesis was assessed using the Amplex® red glucose oxidase assay. The established model was effective in inducing insulin resistance and upregulating gluconeogenesis. Of the tested plant extracts, H. stellipilis showed the most potential as an antidiabetic treatment. It had low toxicity, significantly decreased hepatic glucose production and reduced the amount of phosphoenolpyruvate carboxykinase (PCK) and well as PCK gene expression, and reduced lipid content and reactive oxygen species (ROS) in the C3A cell line. H. stellipilis increased calcium signalling in INS1 rat insulinoma cells, however there was a decrease in expression of genes for insulin and glucose transporter 2 after 6 hr exposure. H. stellipilis appears be beneficial as an antidiabetic treatment. Although antidiabetic studies have been done on other Hypoxis species, this is the first study on the effects of H. stellipilis on gluconeogenesis and diabetes. , Thesis (MSc) -- Faculty of Science, School of Biomolecular & Chemical Sciences, 2023
- Full Text:
- Date Issued: 2023-12
Antiproliferative activity of novel rhenium complexes and medicinal plant extracts
- Oosthuizen, Kenneth Thomas, Venables, Luanne
- Authors: Oosthuizen, Kenneth Thomas , Venables, Luanne
- Date: 2021-04
- Subjects: Gqeberha (South Africa) , Eastern Cape (South Africa) , Medicinal plants -- South Africa
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10948/52028 , vital:43441
- Description: Cancer is a complex, multifactorial disease that affects millions of individuals every year. The adverse side effects and escalating costs of current therapies coupled with the increased incidence of resistance to these therapies make it imperative that we explore novel treatments for the disease. This study investigated two avenues for novel drug design namely, novel synthetic compounds and medicinal plant extracts. The benzimidazole ring system has shown potential as a scaffold for designing novel anticancer agents. Conjugation of the metal rhenium to novel variants of this ring system open up the possibility of designing novel drugs that serve both a diagnostic and a therapeutic function. This study investigated the in vitro anticancer potential of ten such complexes against selected breast and cervical cancer cell lines. The selectivity of the complexes for cancer cells over normal cells was also investigated while the mechanism of action of effective complexes was determined by exploring cell cycle arrest, biochemical markers of apoptosis and mitochondrial membrane disruption. All ten complexes were screened against MCF7 breast and HeLa cervical cancer cell lines with four showing antiproliferative activity against both cancer cell lines and one showing cell line specific toxicity against MCF7 breast cancer cells. When looking at the SAR of the compounds it was noted that activity was higher in compounds which contain two potentially bidentate benzothiazole ligands, while compounds with potentially tridentate ligands show good activity, but only if the benzothiazole moiety is not involved in coordination. The complexes showed greater antiproliferative activity against the MCF7 breast cancer cells with IC50 values ranging from 3.2 to 7.0 µM versus a range of 7.6 to 24.4 µM being obtained on the HeLa cervical cancer cells. Antiproliferative complexes were tested against confluent and log phase Vero cells at their respective IC50 values to determine their effect on “normal” cells. Confluent Vero cells showed less cell death than those in log phase indicating that the complexes show preference for proliferating cells. The mechanism of action of the complexes was studied on both cancer cell lines via cell cycle analysis and apoptosis assays investigating phosphatidylserine translocation, caspase -3 and -8 activation and mitochondrial membrane potential with most complexes inducing cell cycle arrest followed by cell death via both the extrinsic and intrinsic pathways of apoptosis. Since 1940, 49% of all the available anticancer drugs approved for cancer treatment were natural products or directly derived from natural products. Plants are an excellent source of secondary metabolites, many of which are unique chemical compounds that cannot be synthesized in a laboratory. Ethnobotanical surveys conducted in conjunction with Traditional Health Practitioners of the Mkuranga and Same districts in Tanzania identified 25 plants that are used for the treatment of cancer. Four of these plants (A. mossambicensis, C. adenocaule, C. pseudopulchelusis and R. natalensis) with ethnobotanically reported anticancer usage showed cytotoxic activity against brine shrimp and were selected for further in vitro anticancer studies. All four plants were found to have antiproliferative activity against HeLa cervical cancer cells with IC50 values ranging from 3.4 to 50.8 µg/mL. This study was also tasked with investigating the mechanism of action of C. pseudopulchelus on HeLa cervical cancer cells by exploring cell cycle arrest, biochemical markers of apoptosis and mitochondrial membrane x disruption. C. pseudopulchelus caused early M phase arrest followed by slippage and subsequent cell death via the extrinsic pathway of apoptosis. In conclusion, this study showed that both novel complexes as well as medicinal plant extracts represent an interesting avenue for the development of novel anticancer drugs that are cheaper and produce less side effects than current therapies. , Thesis (MSc) -- Faculty of Science, School of Biomolecular and Chemical Sciences, 2021
- Full Text: false
- Date Issued: 2021-04
- Authors: Oosthuizen, Kenneth Thomas , Venables, Luanne
- Date: 2021-04
- Subjects: Gqeberha (South Africa) , Eastern Cape (South Africa) , Medicinal plants -- South Africa
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10948/52028 , vital:43441
- Description: Cancer is a complex, multifactorial disease that affects millions of individuals every year. The adverse side effects and escalating costs of current therapies coupled with the increased incidence of resistance to these therapies make it imperative that we explore novel treatments for the disease. This study investigated two avenues for novel drug design namely, novel synthetic compounds and medicinal plant extracts. The benzimidazole ring system has shown potential as a scaffold for designing novel anticancer agents. Conjugation of the metal rhenium to novel variants of this ring system open up the possibility of designing novel drugs that serve both a diagnostic and a therapeutic function. This study investigated the in vitro anticancer potential of ten such complexes against selected breast and cervical cancer cell lines. The selectivity of the complexes for cancer cells over normal cells was also investigated while the mechanism of action of effective complexes was determined by exploring cell cycle arrest, biochemical markers of apoptosis and mitochondrial membrane disruption. All ten complexes were screened against MCF7 breast and HeLa cervical cancer cell lines with four showing antiproliferative activity against both cancer cell lines and one showing cell line specific toxicity against MCF7 breast cancer cells. When looking at the SAR of the compounds it was noted that activity was higher in compounds which contain two potentially bidentate benzothiazole ligands, while compounds with potentially tridentate ligands show good activity, but only if the benzothiazole moiety is not involved in coordination. The complexes showed greater antiproliferative activity against the MCF7 breast cancer cells with IC50 values ranging from 3.2 to 7.0 µM versus a range of 7.6 to 24.4 µM being obtained on the HeLa cervical cancer cells. Antiproliferative complexes were tested against confluent and log phase Vero cells at their respective IC50 values to determine their effect on “normal” cells. Confluent Vero cells showed less cell death than those in log phase indicating that the complexes show preference for proliferating cells. The mechanism of action of the complexes was studied on both cancer cell lines via cell cycle analysis and apoptosis assays investigating phosphatidylserine translocation, caspase -3 and -8 activation and mitochondrial membrane potential with most complexes inducing cell cycle arrest followed by cell death via both the extrinsic and intrinsic pathways of apoptosis. Since 1940, 49% of all the available anticancer drugs approved for cancer treatment were natural products or directly derived from natural products. Plants are an excellent source of secondary metabolites, many of which are unique chemical compounds that cannot be synthesized in a laboratory. Ethnobotanical surveys conducted in conjunction with Traditional Health Practitioners of the Mkuranga and Same districts in Tanzania identified 25 plants that are used for the treatment of cancer. Four of these plants (A. mossambicensis, C. adenocaule, C. pseudopulchelusis and R. natalensis) with ethnobotanically reported anticancer usage showed cytotoxic activity against brine shrimp and were selected for further in vitro anticancer studies. All four plants were found to have antiproliferative activity against HeLa cervical cancer cells with IC50 values ranging from 3.4 to 50.8 µg/mL. This study was also tasked with investigating the mechanism of action of C. pseudopulchelus on HeLa cervical cancer cells by exploring cell cycle arrest, biochemical markers of apoptosis and mitochondrial membrane x disruption. C. pseudopulchelus caused early M phase arrest followed by slippage and subsequent cell death via the extrinsic pathway of apoptosis. In conclusion, this study showed that both novel complexes as well as medicinal plant extracts represent an interesting avenue for the development of novel anticancer drugs that are cheaper and produce less side effects than current therapies. , Thesis (MSc) -- Faculty of Science, School of Biomolecular and Chemical Sciences, 2021
- Full Text: false
- Date Issued: 2021-04
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