In vitro investigation of the anti-hyperglycemic and chemical composition of Heteromorpha arborescens (Spreng.) Cham leaf extracts used in the management of diabetes mellitus
- Abifarin, Taiwo Oluwafunmilola https://orcid.org/00000003-4994-4638
- Authors: Abifarin, Taiwo Oluwafunmilola https://orcid.org/00000003-4994-4638
- Date: 2021-09
- Subjects: Diabetes
- Language: English
- Type: Doctoral theses , text
- Identifier: http://hdl.handle.net/10353/22798 , vital:52760
- Description: Diabetes mellitus is currently a major threat all over the world. It is a chronic disorder characterized by elevated blood glucose levels and disturbance in carbohydrate, fat and protein metabolism which often times leads to several complications such as, atherosclerosis, diabetic nephropathy, retinopathy and neuropathy. Due to the side effects associated with synthetic oral hypoglycemic drugs, herbal remedies have become the alternative in the management of diabetes mellitus. Heteromorpha arborescens is one of the many plants used as herbal remedies for the management of diabetes mellitus in South Africa. In addition, the roots, barks and leaves are used in other parts of Southern Africa for the treatment of mental problems, asthma, cough, dysentery and tuberculosis. The roots are fed to malnourished children in Botswana and Swaziland. However, despite the high medicinal importance, there is little or no scientificinformation to prove this claim. Therefore, the basis of this study is to determine the chemical composition as well as perform and invitro investigation of the antihyperglycemic potential of H. arborescens leaf extracts to provide information that could validate the ethno-medicinal claims for the use of this plant in the management of diabetes mellitus. Phytochemical contents and antioxidant activity of the leaf extracts were determined. Phytochemical analysis of the acetone, ethanol, aqueous and blanched extracts of H. arborescens leaves indicated that the total phenol content of the extracts ranged between 15.10 mg GAE/g- 42.50 mg QAE/g, proanthocyanidin, 459-8402.1 mg QE/g and the flavonoid content, 109.24- 235.79 mg QE/g. In addition, alkaloids (7.65percent) and saponin (25.33percent) were present in significantamounts. Based on the IC50 values, the ethanol extract exhibited the highest total antioxidant activity (0.0125 mg/mL) with highest inhibition against DPPH and ABTS radicals (0.06 and 0.049 mg/mL respectively). Relatively high antioxidant activity may be attributed to the polyphenolic contents which possess hypoglycemic potentials. Minimum Inhibitory Concentrations (MIC) of H. arborescens leaf extracts against Bacillus pumilus, Staphylococcus epidermidis, Staphylococcus aureus, Escherichia coli and Klebselia pneumoniae was also determined and results indicated considerable antibacterial activity in the acetone, ethanol and blanched extracts with MIC values ranging from 1.563-12.5 mg/mL; however, the aqueous extract was inactive against all the bacteria strains. The antibacterial activity of the leaf extracts could be advantageous against diabetic related infections. The cytotoxicity, anti-obesity and antidiabetic potentials of blanched, aqueous and ethanol extracts of Heteromorpha arborescens (Spreng.) Cham leaves were also investigated. The results revealed that both ethanol and aqueous extracts indicated considerable inhibition against α-glucosidase (IC50 of 627.29 ± 4.33 μg/mL and 576.46 ± 3.21 μg/mL respectively) while the blanched extract showed weak α- glucosidase inhibition (IC50; 855.38 ± 4.29 μg/mL). The aqueous extract showed the best α- amylase inhibition (IC50; 583.74 ± 5.87 μg/mL) among the assessed extracts. However, weak α- amylase inhibition was observed in the ethanol (IC50; 724.60 ± 4.33 μg/mL) and blanched extracts (IC50; 791.63 ± 3.76 μg/mL) of H. arborescens leaves. Some level of glucose utilization in both C3A and L6 cells was also observed for the aqueous and ethanol extracts which may be attributed to the relatively lower toxicity levels present in them, however, glucose utilization was very weak for the blanched extract, which may be due to higher level of cytotoxicity it possessed. Relatively weak lipase inhibition was observed for the ethanol (IC50; 699.3 ± 1.33 μg/mL), aqueous (IC50; 811.52 ± 3.52 μg/mL) and blanched extract (IC50; 1152.7 ± 4.61 μg/mL) as compared to orlistat (IC50; 56.88 ± 0.11 μg/mL). However, there was no reasonable reduction in lipid accumulation observed in all the extract treated cells. The results obtained suggest that H.arborescens leaf extracts can serve as a potential tool for the development of new strategies for the treatment of diabetes and obesity. However, further analysis is required to ascertain its anti- obesity potential. Also, caution should be taken in the use of the plant at high concentrations in order to ensure safety and efficacy. Analyses were also carried out to determine the nutritional and antinutritional constituents of the plant for possible inclusion in the diet of diabetic patients. Proximate analysis revealed the presence of 8.5 percent total ash, 4.92 percent crude fat, 8.41 percent moisture, 15.74 percent crude protein, 21.48 percent crude fiber, 40.95 percent carbohydrates and 271.04 kcal/100g energy value. Mineral analysis showed that H. arborescens leaves are very rich in K, Ca, and Fe. Considerable amounts of Mg, Mn, Na, P, Cu and Zn were also present. Vitamin analysis showed that the plant has a high content of vitamins A, C and E. The anti-nutrients evaluated were phytate, oxalate, saponin, and alkaloids, all of which were below toxic levels except for saponin which was observed in moderately high level. This study revealed that H. arborescens leaves are a good source of nutrients and mineral elements, (with low anti-nutrient content) that are highly beneficial to human health especially in diabetic individuals, therefore, encouraging its possible inclusion as a vegetable. Essential oil composition of fresh Heteromorphaarborescens leaves were also determined by Solvent-Free-Microwave-Extraction (SFME) and Hydrodistillation (HD) methods and the compositions of both mehods were compared in terms of their chemical compositions, yield, CO2 emission and energy consumption. Solvent Free Microwave extraction method indicated higher oil yield of 0.7 mL/200 g (0.35 percent) as compared to 0.59 mL/200 g (0.295 percent), lower energy consumption and CO2 emission as compared to the hydrodistillation method. In conclusion, H. arborescens leaves indicated considerable potential efficacy in the management for diabetes mellitus and may require further structural elucidation and characterization in order to identify the bioactive constituents. , Thesis (PhD) -- Faculty of Science and Agriculture, 2021
- Full Text:
- Authors: Abifarin, Taiwo Oluwafunmilola https://orcid.org/00000003-4994-4638
- Date: 2021-09
- Subjects: Diabetes
- Language: English
- Type: Doctoral theses , text
- Identifier: http://hdl.handle.net/10353/22798 , vital:52760
- Description: Diabetes mellitus is currently a major threat all over the world. It is a chronic disorder characterized by elevated blood glucose levels and disturbance in carbohydrate, fat and protein metabolism which often times leads to several complications such as, atherosclerosis, diabetic nephropathy, retinopathy and neuropathy. Due to the side effects associated with synthetic oral hypoglycemic drugs, herbal remedies have become the alternative in the management of diabetes mellitus. Heteromorpha arborescens is one of the many plants used as herbal remedies for the management of diabetes mellitus in South Africa. In addition, the roots, barks and leaves are used in other parts of Southern Africa for the treatment of mental problems, asthma, cough, dysentery and tuberculosis. The roots are fed to malnourished children in Botswana and Swaziland. However, despite the high medicinal importance, there is little or no scientificinformation to prove this claim. Therefore, the basis of this study is to determine the chemical composition as well as perform and invitro investigation of the antihyperglycemic potential of H. arborescens leaf extracts to provide information that could validate the ethno-medicinal claims for the use of this plant in the management of diabetes mellitus. Phytochemical contents and antioxidant activity of the leaf extracts were determined. Phytochemical analysis of the acetone, ethanol, aqueous and blanched extracts of H. arborescens leaves indicated that the total phenol content of the extracts ranged between 15.10 mg GAE/g- 42.50 mg QAE/g, proanthocyanidin, 459-8402.1 mg QE/g and the flavonoid content, 109.24- 235.79 mg QE/g. In addition, alkaloids (7.65percent) and saponin (25.33percent) were present in significantamounts. Based on the IC50 values, the ethanol extract exhibited the highest total antioxidant activity (0.0125 mg/mL) with highest inhibition against DPPH and ABTS radicals (0.06 and 0.049 mg/mL respectively). Relatively high antioxidant activity may be attributed to the polyphenolic contents which possess hypoglycemic potentials. Minimum Inhibitory Concentrations (MIC) of H. arborescens leaf extracts against Bacillus pumilus, Staphylococcus epidermidis, Staphylococcus aureus, Escherichia coli and Klebselia pneumoniae was also determined and results indicated considerable antibacterial activity in the acetone, ethanol and blanched extracts with MIC values ranging from 1.563-12.5 mg/mL; however, the aqueous extract was inactive against all the bacteria strains. The antibacterial activity of the leaf extracts could be advantageous against diabetic related infections. The cytotoxicity, anti-obesity and antidiabetic potentials of blanched, aqueous and ethanol extracts of Heteromorpha arborescens (Spreng.) Cham leaves were also investigated. The results revealed that both ethanol and aqueous extracts indicated considerable inhibition against α-glucosidase (IC50 of 627.29 ± 4.33 μg/mL and 576.46 ± 3.21 μg/mL respectively) while the blanched extract showed weak α- glucosidase inhibition (IC50; 855.38 ± 4.29 μg/mL). The aqueous extract showed the best α- amylase inhibition (IC50; 583.74 ± 5.87 μg/mL) among the assessed extracts. However, weak α- amylase inhibition was observed in the ethanol (IC50; 724.60 ± 4.33 μg/mL) and blanched extracts (IC50; 791.63 ± 3.76 μg/mL) of H. arborescens leaves. Some level of glucose utilization in both C3A and L6 cells was also observed for the aqueous and ethanol extracts which may be attributed to the relatively lower toxicity levels present in them, however, glucose utilization was very weak for the blanched extract, which may be due to higher level of cytotoxicity it possessed. Relatively weak lipase inhibition was observed for the ethanol (IC50; 699.3 ± 1.33 μg/mL), aqueous (IC50; 811.52 ± 3.52 μg/mL) and blanched extract (IC50; 1152.7 ± 4.61 μg/mL) as compared to orlistat (IC50; 56.88 ± 0.11 μg/mL). However, there was no reasonable reduction in lipid accumulation observed in all the extract treated cells. The results obtained suggest that H.arborescens leaf extracts can serve as a potential tool for the development of new strategies for the treatment of diabetes and obesity. However, further analysis is required to ascertain its anti- obesity potential. Also, caution should be taken in the use of the plant at high concentrations in order to ensure safety and efficacy. Analyses were also carried out to determine the nutritional and antinutritional constituents of the plant for possible inclusion in the diet of diabetic patients. Proximate analysis revealed the presence of 8.5 percent total ash, 4.92 percent crude fat, 8.41 percent moisture, 15.74 percent crude protein, 21.48 percent crude fiber, 40.95 percent carbohydrates and 271.04 kcal/100g energy value. Mineral analysis showed that H. arborescens leaves are very rich in K, Ca, and Fe. Considerable amounts of Mg, Mn, Na, P, Cu and Zn were also present. Vitamin analysis showed that the plant has a high content of vitamins A, C and E. The anti-nutrients evaluated were phytate, oxalate, saponin, and alkaloids, all of which were below toxic levels except for saponin which was observed in moderately high level. This study revealed that H. arborescens leaves are a good source of nutrients and mineral elements, (with low anti-nutrient content) that are highly beneficial to human health especially in diabetic individuals, therefore, encouraging its possible inclusion as a vegetable. Essential oil composition of fresh Heteromorphaarborescens leaves were also determined by Solvent-Free-Microwave-Extraction (SFME) and Hydrodistillation (HD) methods and the compositions of both mehods were compared in terms of their chemical compositions, yield, CO2 emission and energy consumption. Solvent Free Microwave extraction method indicated higher oil yield of 0.7 mL/200 g (0.35 percent) as compared to 0.59 mL/200 g (0.295 percent), lower energy consumption and CO2 emission as compared to the hydrodistillation method. In conclusion, H. arborescens leaves indicated considerable potential efficacy in the management for diabetes mellitus and may require further structural elucidation and characterization in order to identify the bioactive constituents. , Thesis (PhD) -- Faculty of Science and Agriculture, 2021
- Full Text:
Investigation of antidiabetic properties, mechanisms of action and toxicology of Strychnos Henningsii (GILG) bark
- Authors: Oyewole, Oyedemi Sunday
- Date: 2011
- Subjects: Medicinal plants -- South Africa -- Eastern Cape , Traditional medicine -- South Africa -- Eastern Cape , Diabetics -- Alternative treatment -- South Africa , Loganiaceae , Lamiaceae , Diabetes , Proteins
- Language: English
- Type: Thesis , Doctoral , PhD (Biochemistry)
- Identifier: vital:11254 , http://hdl.handle.net/10353/d1001070 , Medicinal plants -- South Africa -- Eastern Cape , Traditional medicine -- South Africa -- Eastern Cape , Diabetics -- Alternative treatment -- South Africa , Loganiaceae , Lamiaceae , Diabetes , Proteins
- Description: The apparent reversal of trend from modern drugs to herbal medicine is partly due to the fact that synthetic drugs have always shown adverse reactions and other undesirable side effects. Hence, the use of medicinal plants for the treatment of diseases such as diabetes is very common especially in the rural areas. Majority of these plants are used based on the experience and indigenous knowledge without identification of the therapeutic agents. There is enormous wealth of medicinal plants in the world yet many of them have not been discovered or studied scientifically to substantiate their ethno-medicinal usages. Ethnobotanical study has been the method often used to search for locally important plant species for the discovery of crude drugs with low side effects. An ethnobotanical survey was conducted on the medicinal plants commonly used for the management of diabetes mellitus in Nkonkobe Municipality, Eastern Cape of South Africa. Information was obtained through structured questionnaire administered to traditional healers and herbalists in the region. The study revealed 15 species of plants belonging to 13 families. Strychnos henningsii and Leonotis leonorus among others were repeatedly mentioned by the traditional healers as the two mostly used plants for the management of diabetes mellitus. The infusion and decoction of the roots, leaves and barks of these plants are the methods of preparation. The antioxidant potential of aqueous bark extract of S. henningsii was investigated both in vivo and in vitro using spectroscopic method. The antioxidant activity of the extract against hydrogen peroxide (H2O2), 2,2′-azinobis[3-ethylbenzothiazoline6-sulfonic acid] diammonium salt (ABTS), as well as reducing power was concentration dependent. The extract exhibited lower and average scavenging activities against 1,1diphenyl2picrylhydrazyl (DPPH) and nitric oxide (NO) radicals with IC50 value of 0.739 and 0.49 mg/ml respectively. The administration of the plant extract at 250, 500 and 1000 mg/kg significantly increased the activities of the antioxidant enzymes in the hepatotoxic rats induced with carbon tetrachloride. On the other hand, the stem bark extract had lower effect on lipid peroxidation level except at the dose of 250 mg/kg. The effect of oral administration of S. henningsii extract was evaluated in normal Wistar rats for 28 days. The observed result indicated non- toxic effect of sub-acute administration of plant extract to the animals except at certain doses. This is because, there was no apparent damage to some haematological and biochemical parameters used in assessing organ specific toxicity. However, the alterations observed on platelet, white blood cells and its differentials imply parameter and dose selective toxicity when repeatedly consumed on daily basis at the doses investigated. This study also investigated the antidiabetic activities of the extract at the doses of 125, 250 and 500 mg/kg body weight in diabetic rats induced with streptozotocin -nicotinamide for 15 days. The extract appreciably (P <0.05) reduced the blood glucose level, feed and water intake while the best result was obtained at 250 mg/kg. Similarly, the level of triacylglycerol at the three doses investigated was significantly decreased. In addition, the glucose tolerance was reduced to near normal level after 90 min at certain doses. The clinical significance of the extract on some biochemical and haematological parameters lessen both hepatic and renal damages. Anaemic condition in diabetic animals was also improved after plant extract administration. However, no significant effect was observed in white blood cells and some of its differentials. The extract demonstrated strong glucose utilization in 3T3-L1 cells with a response of 278.63 percent of the control at 12.5μg/ml while that of Chang liver cells was 103.54 percent. The cytotoxicity result revealed non toxic effects of the extract to both cell lines. Treatment of 3T3 L1 cells with the extract did not reduce lipid accumulation. The extract inhibited the activity of α- glucosidase and α- amylase in a concentration dependent manner with IC50 values of 38 μg/ml and 60.9 μg/ml respectively. The percentage protein antiglycation of S. henningsii was 18.4, 38.2 and 61.2 perceent for 0.25, 0.5 and 1 mg/ml respectively while aminoguanidine a known inhibitor of protein glycation was 87.2 percent at 1 mg/ml. The FRAP assay values of the extract was 357.05 μmol Fe (II)/g. The findings from this study support the folkloric usage of this plant for the management of diabetes mellitus in the region.
- Full Text:
- Authors: Oyewole, Oyedemi Sunday
- Date: 2011
- Subjects: Medicinal plants -- South Africa -- Eastern Cape , Traditional medicine -- South Africa -- Eastern Cape , Diabetics -- Alternative treatment -- South Africa , Loganiaceae , Lamiaceae , Diabetes , Proteins
- Language: English
- Type: Thesis , Doctoral , PhD (Biochemistry)
- Identifier: vital:11254 , http://hdl.handle.net/10353/d1001070 , Medicinal plants -- South Africa -- Eastern Cape , Traditional medicine -- South Africa -- Eastern Cape , Diabetics -- Alternative treatment -- South Africa , Loganiaceae , Lamiaceae , Diabetes , Proteins
- Description: The apparent reversal of trend from modern drugs to herbal medicine is partly due to the fact that synthetic drugs have always shown adverse reactions and other undesirable side effects. Hence, the use of medicinal plants for the treatment of diseases such as diabetes is very common especially in the rural areas. Majority of these plants are used based on the experience and indigenous knowledge without identification of the therapeutic agents. There is enormous wealth of medicinal plants in the world yet many of them have not been discovered or studied scientifically to substantiate their ethno-medicinal usages. Ethnobotanical study has been the method often used to search for locally important plant species for the discovery of crude drugs with low side effects. An ethnobotanical survey was conducted on the medicinal plants commonly used for the management of diabetes mellitus in Nkonkobe Municipality, Eastern Cape of South Africa. Information was obtained through structured questionnaire administered to traditional healers and herbalists in the region. The study revealed 15 species of plants belonging to 13 families. Strychnos henningsii and Leonotis leonorus among others were repeatedly mentioned by the traditional healers as the two mostly used plants for the management of diabetes mellitus. The infusion and decoction of the roots, leaves and barks of these plants are the methods of preparation. The antioxidant potential of aqueous bark extract of S. henningsii was investigated both in vivo and in vitro using spectroscopic method. The antioxidant activity of the extract against hydrogen peroxide (H2O2), 2,2′-azinobis[3-ethylbenzothiazoline6-sulfonic acid] diammonium salt (ABTS), as well as reducing power was concentration dependent. The extract exhibited lower and average scavenging activities against 1,1diphenyl2picrylhydrazyl (DPPH) and nitric oxide (NO) radicals with IC50 value of 0.739 and 0.49 mg/ml respectively. The administration of the plant extract at 250, 500 and 1000 mg/kg significantly increased the activities of the antioxidant enzymes in the hepatotoxic rats induced with carbon tetrachloride. On the other hand, the stem bark extract had lower effect on lipid peroxidation level except at the dose of 250 mg/kg. The effect of oral administration of S. henningsii extract was evaluated in normal Wistar rats for 28 days. The observed result indicated non- toxic effect of sub-acute administration of plant extract to the animals except at certain doses. This is because, there was no apparent damage to some haematological and biochemical parameters used in assessing organ specific toxicity. However, the alterations observed on platelet, white blood cells and its differentials imply parameter and dose selective toxicity when repeatedly consumed on daily basis at the doses investigated. This study also investigated the antidiabetic activities of the extract at the doses of 125, 250 and 500 mg/kg body weight in diabetic rats induced with streptozotocin -nicotinamide for 15 days. The extract appreciably (P <0.05) reduced the blood glucose level, feed and water intake while the best result was obtained at 250 mg/kg. Similarly, the level of triacylglycerol at the three doses investigated was significantly decreased. In addition, the glucose tolerance was reduced to near normal level after 90 min at certain doses. The clinical significance of the extract on some biochemical and haematological parameters lessen both hepatic and renal damages. Anaemic condition in diabetic animals was also improved after plant extract administration. However, no significant effect was observed in white blood cells and some of its differentials. The extract demonstrated strong glucose utilization in 3T3-L1 cells with a response of 278.63 percent of the control at 12.5μg/ml while that of Chang liver cells was 103.54 percent. The cytotoxicity result revealed non toxic effects of the extract to both cell lines. Treatment of 3T3 L1 cells with the extract did not reduce lipid accumulation. The extract inhibited the activity of α- glucosidase and α- amylase in a concentration dependent manner with IC50 values of 38 μg/ml and 60.9 μg/ml respectively. The percentage protein antiglycation of S. henningsii was 18.4, 38.2 and 61.2 perceent for 0.25, 0.5 and 1 mg/ml respectively while aminoguanidine a known inhibitor of protein glycation was 87.2 percent at 1 mg/ml. The FRAP assay values of the extract was 357.05 μmol Fe (II)/g. The findings from this study support the folkloric usage of this plant for the management of diabetes mellitus in the region.
- Full Text:
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