In silico and in vitro screening of marrubiin and marrubiin derivatives for antidiabetic activity on PTP1ß, C2C12 myocytes, chang liver hepatocytes and 3T3-L1 adipocytes
- Authors: Nicholas, Rudi Berto
- Date: 2013
- Subjects: Hypoglycemic agents , Diabetes -- Treatment -- South Africa
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10347 , http://hdl.handle.net/10948/d1020638
- Description: Diabetes mellitus (DM) is a life changing disease which affects a large portion of the population and the economy through high medical costs and loss of productivity. Marrubiin (MAR), a diterpenoid isolated from Leonotis leonurus, a plant indigenous to Southern Africa, is used by traditional healers to alleviate DM symptoms. This study aims to screen the inhibitory potential of MAR and MAR derivatives on PTP1β and glucose uptake properties of Chang liver, C2C12 and 3T3-L1 cells. Marrubiin and 19 of its derivatives were tested to determine the inhibition constants for PTP1β. A Ki of 21 μM and 0.047 μM was detected for oleanolic acid in silico and in vitro, respectively. All other diterpene derivatives did not display substantial levels of inhibition of PTP1β. Treatment of Chang liver cells with the various MAR derivatives (10 μM) did not significantly increase glucose uptake beyond metformin, a known antidiabetic drug. The various treatments showed a protective/proliferative effect on the C2C12 muscle cells with two MAR treatments (DC16 and DC18) significantly increasing glucose uptake as compared to metformin in C2C12 muscle cells. It was noted that DC17, DC18 and MAR significantly increased glucose uptake in 3T3-L1 adipocytes, relative to the control. Contrary to cytotoxicity studies with Chang liver and C2C12 muscle cells, adipocytes displayed no cytotoxicity to treatments while a significant increase in cell viability was seen for DC9 and DC15. To unravel the mechanism of action, Western blotting analysis was completed and an increased expression of PTP1β was observed for treatments with DC17 and DC6 was seen in adipocytes, while DC18 and metformin decreased expression significantly. This correlated with a significant decrease of Ser 612 phosphorylation of insulin receptor substrate (IRS1) for DC17. Real time qPCR of IRS1 and GLUT4 highlighted that DC17 and MAR were able to significantly increase expression of IRS1 and GLUT4, respectively. The results show that MAR and the selected derivatives (DC6, DC17, DC18) have been found to increase glucose uptake in peripheral tissue types with IRS1, GLUT4 and PTP1β being associated with the mechanism of action. However, a complete understanding of the mechanisms is yet to be established.
- Full Text:
- Date Issued: 2013
- Authors: Nicholas, Rudi Berto
- Date: 2013
- Subjects: Hypoglycemic agents , Diabetes -- Treatment -- South Africa
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10347 , http://hdl.handle.net/10948/d1020638
- Description: Diabetes mellitus (DM) is a life changing disease which affects a large portion of the population and the economy through high medical costs and loss of productivity. Marrubiin (MAR), a diterpenoid isolated from Leonotis leonurus, a plant indigenous to Southern Africa, is used by traditional healers to alleviate DM symptoms. This study aims to screen the inhibitory potential of MAR and MAR derivatives on PTP1β and glucose uptake properties of Chang liver, C2C12 and 3T3-L1 cells. Marrubiin and 19 of its derivatives were tested to determine the inhibition constants for PTP1β. A Ki of 21 μM and 0.047 μM was detected for oleanolic acid in silico and in vitro, respectively. All other diterpene derivatives did not display substantial levels of inhibition of PTP1β. Treatment of Chang liver cells with the various MAR derivatives (10 μM) did not significantly increase glucose uptake beyond metformin, a known antidiabetic drug. The various treatments showed a protective/proliferative effect on the C2C12 muscle cells with two MAR treatments (DC16 and DC18) significantly increasing glucose uptake as compared to metformin in C2C12 muscle cells. It was noted that DC17, DC18 and MAR significantly increased glucose uptake in 3T3-L1 adipocytes, relative to the control. Contrary to cytotoxicity studies with Chang liver and C2C12 muscle cells, adipocytes displayed no cytotoxicity to treatments while a significant increase in cell viability was seen for DC9 and DC15. To unravel the mechanism of action, Western blotting analysis was completed and an increased expression of PTP1β was observed for treatments with DC17 and DC6 was seen in adipocytes, while DC18 and metformin decreased expression significantly. This correlated with a significant decrease of Ser 612 phosphorylation of insulin receptor substrate (IRS1) for DC17. Real time qPCR of IRS1 and GLUT4 highlighted that DC17 and MAR were able to significantly increase expression of IRS1 and GLUT4, respectively. The results show that MAR and the selected derivatives (DC6, DC17, DC18) have been found to increase glucose uptake in peripheral tissue types with IRS1, GLUT4 and PTP1β being associated with the mechanism of action. However, a complete understanding of the mechanisms is yet to be established.
- Full Text:
- Date Issued: 2013
The South African community pharmacist and Type 2 Diabetes Mellitus a pharmaceutical care intervention
- Authors: Hill, Peter William
- Date: 2009
- Subjects: Pharmacist and patient -- South Africa , Pharmaceutical services -- Patients , Pharmaceutical services -- South Africa , Pharmacists -- South Africa , Diabetes -- Treatment -- South Africa , Community health services -- South Africa
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:3760 , http://hdl.handle.net/10962/d1003238 , Pharmacist and patient -- South Africa , Pharmaceutical services -- Patients , Pharmaceutical services -- South Africa , Pharmacists -- South Africa , Diabetes -- Treatment -- South Africa , Community health services -- South Africa
- Description: Type 2 diabetes mellitus is a chronic disease of pandemic magnitude, increasingly contributing to the disease burden of countries in the developing world, largely because of the effects of unhealthy lifestyles fuelled by unbridled urbanisation. In certain settings, patients with diabetes are more likely to have a healthcare encounter with a pharmacist than with any other healthcare provider. The overall aim of the study was to investigate the potential of South African community pharmacists to positively influence patient adherence and metabolic control in Type 2 diabetes. The designated primary endpoint was glycated haemoglobin, with the intermediate health outcomes of blood lipids, serum creatinine, blood pressure and body mass index serving as secondary endpoints. Community pharmacists and their associated Type 2 diabetes patients were recruited from areas throughout South Africa using the communication media of various nonstatutory pharmacy organisations. Although 156 pharmacists initially indicated interest in participating in the study, only 28 pharmacists and 153 patients were enrolled prior to baseline data collection. Of these, 16 pharmacists and 57 patients participated in the study for the full twelve months. Baseline clinical and psychosocial data were collected, after which pharmacists and their patients were randomised, nine pharmacists and 34 patients to the intervention group and 8 pharmacists and 27 patients to the control group. The sample size calculation revealed that each group required the participation of a minimum of 35 patients. Control pharmacists were requested to offer standard pharmaceutical care, while the intervention pharmacists were provided with a scope of practice diabetes care plan to guide the diabetes care they were to provide. Data were again collected 12-months postbaseline. At baseline, proportionally more intervention patients (82.4%) than control patients (59.3%) were using only oral anti-diabetes agents (i.e. not in combination with insulin), while insulin usage, either alone or in combination with oral agents was conversely greater in the control group (40.7%) than in the intervention group (17.6%) (Chi-squared test, p=0.013). Approximately half of the patients (53.8% control and 47.1% intervention) reported having their HbA1c levels measured in terms of accepted guidelines. There was no significant difference in HbA1c between the groups at the end of the study (Independent t-test, p=0.514). In the control group, the mean HbA1c increased from 7.3±1.2% to 7.6±1.5%, while for the intervention patients the variable remained almost constant (8.2±2.0% at baseline and 8.2±1.8% at post-baseline). Similarly, there were no significant differences between the groups with regard to any of the designated secondary clinical endpoints. Adherence to medication and self-management recommendations was similarly good for both groups. There were no significant differences between the two groups for any of the other psychosocial variables measured. In conclusion, intervention pharmacists were not able to significantly influence glycaemic control or therapeutic adherence compared to the control pharmacists.
- Full Text:
- Date Issued: 2009
- Authors: Hill, Peter William
- Date: 2009
- Subjects: Pharmacist and patient -- South Africa , Pharmaceutical services -- Patients , Pharmaceutical services -- South Africa , Pharmacists -- South Africa , Diabetes -- Treatment -- South Africa , Community health services -- South Africa
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:3760 , http://hdl.handle.net/10962/d1003238 , Pharmacist and patient -- South Africa , Pharmaceutical services -- Patients , Pharmaceutical services -- South Africa , Pharmacists -- South Africa , Diabetes -- Treatment -- South Africa , Community health services -- South Africa
- Description: Type 2 diabetes mellitus is a chronic disease of pandemic magnitude, increasingly contributing to the disease burden of countries in the developing world, largely because of the effects of unhealthy lifestyles fuelled by unbridled urbanisation. In certain settings, patients with diabetes are more likely to have a healthcare encounter with a pharmacist than with any other healthcare provider. The overall aim of the study was to investigate the potential of South African community pharmacists to positively influence patient adherence and metabolic control in Type 2 diabetes. The designated primary endpoint was glycated haemoglobin, with the intermediate health outcomes of blood lipids, serum creatinine, blood pressure and body mass index serving as secondary endpoints. Community pharmacists and their associated Type 2 diabetes patients were recruited from areas throughout South Africa using the communication media of various nonstatutory pharmacy organisations. Although 156 pharmacists initially indicated interest in participating in the study, only 28 pharmacists and 153 patients were enrolled prior to baseline data collection. Of these, 16 pharmacists and 57 patients participated in the study for the full twelve months. Baseline clinical and psychosocial data were collected, after which pharmacists and their patients were randomised, nine pharmacists and 34 patients to the intervention group and 8 pharmacists and 27 patients to the control group. The sample size calculation revealed that each group required the participation of a minimum of 35 patients. Control pharmacists were requested to offer standard pharmaceutical care, while the intervention pharmacists were provided with a scope of practice diabetes care plan to guide the diabetes care they were to provide. Data were again collected 12-months postbaseline. At baseline, proportionally more intervention patients (82.4%) than control patients (59.3%) were using only oral anti-diabetes agents (i.e. not in combination with insulin), while insulin usage, either alone or in combination with oral agents was conversely greater in the control group (40.7%) than in the intervention group (17.6%) (Chi-squared test, p=0.013). Approximately half of the patients (53.8% control and 47.1% intervention) reported having their HbA1c levels measured in terms of accepted guidelines. There was no significant difference in HbA1c between the groups at the end of the study (Independent t-test, p=0.514). In the control group, the mean HbA1c increased from 7.3±1.2% to 7.6±1.5%, while for the intervention patients the variable remained almost constant (8.2±2.0% at baseline and 8.2±1.8% at post-baseline). Similarly, there were no significant differences between the groups with regard to any of the designated secondary clinical endpoints. Adherence to medication and self-management recommendations was similarly good for both groups. There were no significant differences between the two groups for any of the other psychosocial variables measured. In conclusion, intervention pharmacists were not able to significantly influence glycaemic control or therapeutic adherence compared to the control pharmacists.
- Full Text:
- Date Issued: 2009
Experiences of the Xhosa diabetic patient
- Authors: Ngamlana, Zodumo Princess
- Date: 2006
- Subjects: Diabetes -- Treatment -- South Africa , Diabetics -- Rehabilitation -- South Africa , Xhosa (African people) -- South Africa
- Language: English
- Type: Thesis , Masters , MCur
- Identifier: vital:10014 , http://hdl.handle.net/10948/465 , Diabetes -- Treatment -- South Africa , Diabetics -- Rehabilitation -- South Africa , Xhosa (African people) -- South Africa
- Description: This study will be focussing on the experiences of Xhosa-speaking patients with DM utilising the NMMM public hospitals complex. In the OPD patients are assessed and treated for all chronic conditions including DM, and patients are seen at monthly intervals or when necessary. The OPD serves the neighbouring black population from the surrounding townships as well as the informal settlements. The effects of urbanisation have resulted in this area having a semi–rural, semi-urban population that is mostly Xhosa speaking. The bulk of the population is unemployed, while others are living on a minimal income. Unemployment in the Eastern Cape ranges from 40% in rural areas, rising to 50 - 60% in the urban areas (Proposed aluminium Pechiney smelter within the Coega IDZ, 2002:4-8). In some homes there is no or little money to buy food, and even less to make use of a health service. Some people live on either a social grant for the elderly, a grant for young children or a disability grant and most people in this area have an income below the level at which payment of taxes for contribution to the economy is possible.
- Full Text:
- Date Issued: 2006
- Authors: Ngamlana, Zodumo Princess
- Date: 2006
- Subjects: Diabetes -- Treatment -- South Africa , Diabetics -- Rehabilitation -- South Africa , Xhosa (African people) -- South Africa
- Language: English
- Type: Thesis , Masters , MCur
- Identifier: vital:10014 , http://hdl.handle.net/10948/465 , Diabetes -- Treatment -- South Africa , Diabetics -- Rehabilitation -- South Africa , Xhosa (African people) -- South Africa
- Description: This study will be focussing on the experiences of Xhosa-speaking patients with DM utilising the NMMM public hospitals complex. In the OPD patients are assessed and treated for all chronic conditions including DM, and patients are seen at monthly intervals or when necessary. The OPD serves the neighbouring black population from the surrounding townships as well as the informal settlements. The effects of urbanisation have resulted in this area having a semi–rural, semi-urban population that is mostly Xhosa speaking. The bulk of the population is unemployed, while others are living on a minimal income. Unemployment in the Eastern Cape ranges from 40% in rural areas, rising to 50 - 60% in the urban areas (Proposed aluminium Pechiney smelter within the Coega IDZ, 2002:4-8). In some homes there is no or little money to buy food, and even less to make use of a health service. Some people live on either a social grant for the elderly, a grant for young children or a disability grant and most people in this area have an income below the level at which payment of taxes for contribution to the economy is possible.
- Full Text:
- Date Issued: 2006
Optimisation of an in vitro model for anti-diabetic screening
- Authors: Wilson, Gayle Pamela
- Date: 2006
- Subjects: Hypoglycemic agents , Diabetes -- Treatment -- South Africa , Materia medica, Vegetable -- South Africa
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10308 , http://hdl.handle.net/10948/428 , Hypoglycemic agents , Diabetes -- Treatment -- South Africa , Materia medica, Vegetable -- South Africa
- Description: The need for alternative strategies for the prevention and treatment of diabetes is growing rapidly as type II diabetes is reaching epidemic status in our society. This need was the basis for the creation of this study, as it was necessary to start looking towards medicinal plants as potential antidiabetic treatment and no comprehensive in vitro model existed. In creating a model for determining the effects of alternative traditional medicines as antidiabetic potentiates, it was necessary that two metabolic pathways, namely glucose uptake and insulin secretion, which play a significant role in glucose homeostasis, be at the centre of our investigations. The objective of this project was to optimize the methodology required to screen and ultimately determine the effectiveness of the plant extracts Kankerbos and MRC2003, as antidiabetic potentiates, through observing their effects on glucose utilisation and insulin secretion. If these medicinal plants are going to make a positive contribution to the health of type II diabetic South Africans, then the determination of their efficacy is essential. The cell lines used in this study included 3T3-L1 preadipocytes, Chang liver, C2C12 muscle and INS-1 rat pancreatic cells. Each cell line represents a different in vivo organ that is known to have an influence on glucose homeostasis in our bodies, each with its own unique metabolic pathways and mechanisms of activity, thereby making each one a vital component in the study. The positive controls for the two models were insulin and metformin (glucose utilisation) and glibenclamide (insulin secretion). Insulin was shown to provide a significant increase in the amount of glucose taken up in C2C12 muscle and Chang liver cells for acute conditions. Chronic treatments with metformin provided a significant increase in glucose utilised by Chang liver cells. Glibenclamide was an effective positive control for stimulating insulin secretion by INS-1 cells under acute conditions as there was a significant increase in the amount of insulin secreted. MRC2003 did not show any significant antidiabetic activity. Sutherlandia frutescens (Kankerbos) showed biological activities comparable to some of the more recognized antidiabetic compounds throughout the study. With regards to the glucose utilisation model, Kankerbos was seen to have both acute and chronic effects in different cell lines. In the C2C12 muscle cell line, Kankerbos significantly increased glucose uptake when they were exposed to acute conditions. Kankerbos also had a significant effect on the Chang liver cells as it was observed that under both acute and chronic conditions, this plant extract induced the uptake of glucose into these cells. With respect to the insulin secretion model involving INS-1 cells, no significant effect was seen during acute exposure with Kankerbos treatment. However during chronic exposure, an increase in insulin secretion was initiated by this plant extract. Overall, the results of this study suggest that Kankerbos has a twofold mechanism of action for its glucose-lowering effects. Given that Kankerbos is widely available in South Africa, this study was valuable as it provided an indication that Kankerbos has antidiabetic activities and could possibly be used as an alternative antidiabetic medication.
- Full Text:
- Date Issued: 2006
- Authors: Wilson, Gayle Pamela
- Date: 2006
- Subjects: Hypoglycemic agents , Diabetes -- Treatment -- South Africa , Materia medica, Vegetable -- South Africa
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10308 , http://hdl.handle.net/10948/428 , Hypoglycemic agents , Diabetes -- Treatment -- South Africa , Materia medica, Vegetable -- South Africa
- Description: The need for alternative strategies for the prevention and treatment of diabetes is growing rapidly as type II diabetes is reaching epidemic status in our society. This need was the basis for the creation of this study, as it was necessary to start looking towards medicinal plants as potential antidiabetic treatment and no comprehensive in vitro model existed. In creating a model for determining the effects of alternative traditional medicines as antidiabetic potentiates, it was necessary that two metabolic pathways, namely glucose uptake and insulin secretion, which play a significant role in glucose homeostasis, be at the centre of our investigations. The objective of this project was to optimize the methodology required to screen and ultimately determine the effectiveness of the plant extracts Kankerbos and MRC2003, as antidiabetic potentiates, through observing their effects on glucose utilisation and insulin secretion. If these medicinal plants are going to make a positive contribution to the health of type II diabetic South Africans, then the determination of their efficacy is essential. The cell lines used in this study included 3T3-L1 preadipocytes, Chang liver, C2C12 muscle and INS-1 rat pancreatic cells. Each cell line represents a different in vivo organ that is known to have an influence on glucose homeostasis in our bodies, each with its own unique metabolic pathways and mechanisms of activity, thereby making each one a vital component in the study. The positive controls for the two models were insulin and metformin (glucose utilisation) and glibenclamide (insulin secretion). Insulin was shown to provide a significant increase in the amount of glucose taken up in C2C12 muscle and Chang liver cells for acute conditions. Chronic treatments with metformin provided a significant increase in glucose utilised by Chang liver cells. Glibenclamide was an effective positive control for stimulating insulin secretion by INS-1 cells under acute conditions as there was a significant increase in the amount of insulin secreted. MRC2003 did not show any significant antidiabetic activity. Sutherlandia frutescens (Kankerbos) showed biological activities comparable to some of the more recognized antidiabetic compounds throughout the study. With regards to the glucose utilisation model, Kankerbos was seen to have both acute and chronic effects in different cell lines. In the C2C12 muscle cell line, Kankerbos significantly increased glucose uptake when they were exposed to acute conditions. Kankerbos also had a significant effect on the Chang liver cells as it was observed that under both acute and chronic conditions, this plant extract induced the uptake of glucose into these cells. With respect to the insulin secretion model involving INS-1 cells, no significant effect was seen during acute exposure with Kankerbos treatment. However during chronic exposure, an increase in insulin secretion was initiated by this plant extract. Overall, the results of this study suggest that Kankerbos has a twofold mechanism of action for its glucose-lowering effects. Given that Kankerbos is widely available in South Africa, this study was valuable as it provided an indication that Kankerbos has antidiabetic activities and could possibly be used as an alternative antidiabetic medication.
- Full Text:
- Date Issued: 2006
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