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ThesisItem Open Access Identification of toxic fractions of Mimosa invisa (Anathottavadi) and its toxicity in rabbits(Department of Pharmacology and Toxicology, College of Veterinary and Animal Sciences, Mannuthy, 2007) Usha, P T A; KAU; Gopakumar, NThe present study was undertaken to identify the toxic fraction of Mimosa invisa in rabbits and to assess to toxicity of Mimosa invisa fresh juice, cold alcoholic extract and various fractions of alcoholic extract utilizing rabbit as a model along with treatment study. The experiment was conducted in two phases. The first part of the study involved assessment of toxicity of fresh juice of Mimosa invisa (group II) and cold alcoholic extract (group Ill). Group I served as control. The preliminary tests were conducted to derive the toxic dose of fresh juice and alcoholic extract of Mimosa invisa. The toxic doses were 25 g/kg and 1 g/kg body weight for Mimosa invisa fresh juice and alcoholic extract respectively. The clinical symptoms, biochemical parameters and haemogram were observed to assess the toxicity. The serum ALT, AST and GGT levels showed significant increase in both the groups. The serum creatine kinase levels exhibited an increase followed by a decrease. There was a significant increase in serum creatinine and urea levels. There were no changes in serum total protein, albumin, globulin and albumin-globulin ratio. Significant decrease in VPRC, haemoglobin and RBC count could be noticed. The erythrocyte indices did not show any variations. The leucocytosis was observed in group II and III when compared to control (group I). Lymphocytosis with neutropenia were also observed in both the groups. The second phase of the study involved identification of toxic dose of each fraction of Mimosa invisa: The preliminary studies revealed that chloroform fraction (Fraction I) and water insoluble residue (fraction IV) were not toxic to rabbits while the fraction II (n-butanol fraction) and fraction III (aqueous fraction) were toxic to rabbits. Hence the two toxic fractions were pooled and used for further studies. It was found that 0.4 g/kg of pooled toxic fraction was toxic in rabbits. The toxicity was assessed by the evaluation of clinical symptoms, biochemical parameters and haemogram. The group V (pooled toxic fraction) showed inappetence, dullness, lethargy and reluctant to move. A significant increase in serum ALT, AST and GGT levels were observed. The serum ALP levels showed an increase followed by a decrease. The serum creatine kinase also showed similar increase followed by decrease. The creatinine and urea levels exhibited a continuous increase in group V. There were no changes in total protein, albumin, globulin and albumin-globulin ratio. The VPRC, RBC and haemoglobin showed significant decrease but there were no changes in erythrocyte indices. A significant leucocytosis was observed in group V. The differential leucocyte count showed lymphocytosis with neutropenia. The group VI (Half the toxic dose of pooled toxic fraction) failed to produce toxicity as evidenced by biochemical parameters and haemogram. The group VII (Double the toxic dose of pooled toxic fraction), all the animals died within 12-24 hours of administration of the dose. The group VIII animals were treated with a decoction prepared from equal quantities of Boerhvia diffusa, Hygrophila auriculata and Tribulus terrestris along with pooled toxic fraction of Mimosa invisa. The prominent symptom of inappetence was only for a short period of time (1-2 day). Then the animals started taking normal feed and water. The serum AL T, AST and GGT levels were significantly increased during the first five days, then the values returned to normal level. The serum creatine kinase and ALP levels did not show significant changes. The serum creatinine and urea levels showed significant increase followed by decrease. All the parameters showed significant improvement when compared with group V (pooled toxic fraction alone). There were no changes in serum protein, 'albumin, globulin and albumin-globulin ratio. The screening of alcoholic extract and various fractions revealed that the alcoholic extract n-butanol fraction and aqueous fraction contained steroids phenolic compounds, tannins, flavonoids, glycosides, diterpenes triterpenes and saponins. The chloroform fraction was positive for flavonoids only, whereas water insoluble fraction contained flavonoids and traces of tannins, triterpenes and saponins. From the results of the present study, it is concluded that the phytotoxin present in M invisa is nephrotoxic and hepatotoxic. The treatment schedule tried using a decoction of Boerhavia diffusa, Tribulus terrestris and Hygrophila auriculata could protect kidney and liver from the phytotoxin present in M invisa. Further study is needed to isolate the phytotoxin present in M invisa so that a specific antidote can be developed.ThesisItem Open Access Hypoglycamic effect of pleurotus ostreatus in sprague - dawley rats(Department of Pharmacology and Toxicology, College of Veterinary and Animal Sciences, Mannuthy, 2007) Saritha, Krishna I K; KAU; Usha, P T AThe present study was undertaken to assess the antidiabetic activity of Pleurotus ostreatus and its combination with Murraya koenigii and Aegle marmelos in a model of alloxan induced diabetic rats. The experiment was conducted in seventy two adult Sprague-Dawley male rats which were randomly divided into nine groups of eight animals each. Group I served as normal control. All the groups except normal control were made diabetic by the subcutaneous injection of alloxan at the dose rate of 130 mg/kg body weight. Group II was kept as the diabetic control. In the initial phase, rats of group III, IV and V were administered orally with ethanolic extract of Pleurotus ostreatus at the dose rate of 250,500 and 1000 mg/kg body weight respectively from day 16 to day 45. Parameters like blood glucose, serum cholesterol and serum triglyceride were estimated on zeroth day, 16th, 30th and 45th day. Body weight was also recorded on these days. Liver glycogen was estimated on 45th day after sacrificing the animals. All the three treatment groups except diabetic control showed gradual increase in body weight after 30 days of treatment. Among the treatment groups, the maximum regain in body weight was observed in rats treated with Pleurotus ostreatus at the dose rate of 1000 mg/kg (group V). The highest decrease in blood glucose level was also observed with the same group. Administration of the extract also produced significant reduction in serum cholesterol level. Group V showed the lowest serum cholesterol level and serum triglyceride level. The liver glycogen level was found to be highest in group V. The results of the present study clearly demonstrated the hypoglycaemic effect of Pleurotus ostreatus. It was also quite evident that among the three doses of Pleurotus ostreatus chosen, the dose rate of 1000 mg/kg was found to possess the highest hypoglycaemic and hypolipidaemic effect. Hence this was selected as the suitable dose for combination with Murraya koenigii and Aegle marmelos. In the second phase of study, group VI received ethanolic extract of Murraya koenigii at the dose rate of 250 mg/kg and group VII was given ethanolic extract of Aegle marmelos at the same dose rate from day 16 to day 45. A combination of the ethanolic extract of Pleurotus ostreatus at the dose rate of 1000 mg/kg with Murraya koenigii and Aegle marmelos each at the dose rate of 250 mg/kg was given to group VIII for the same period. Group IX received glibenclamide at the dose rate of 0.25 mg/kg/day for 30 days. Group I and II served as normal and diabetic control respectively. Parameters like body weight, blood glucose, serum cholesterol and serum triglyceride were recorded on zeroth day, 16th, 30th and 45th day. Liver glycogen was estimated on 45th day after sacrificing the animals. After 30 days of treatment, a significant gain in body weight was observed in animals which underwent combination therapy compared to group VI and VII and the effect was comparable to that produced by glibenclamide. The animals which received Murraya koenigii and Aegle marmelos (groups VI and VII) showed similar reduction in blood glucose level. Group VIII which received combination showed a significant reduction in blood glucose level than groups VI and VII suggesting a synergistic effect of the three drugs in producing hypoglycaemia. However, the most effective reduction in blood glucose level was produced by glibenclamide. A significant decrease in serum cholesterol level was seen in all treated groups compared to diabetic control. Among the groups treated with ethanolic extract, group VIII showed the lowest serum cholesterol level. Group IX which received glibenclamide showed the highest reduction in serum cholesterol level. The highest reduction in serum triglyceride level was also shown by group IX followed by group VIII. Animals treated with Aegle marmelos extract (group VII) had a higher liver glycogen value than those treated with Murraya koenigii (group VI). Treatment with the combination produced significant regain in liver glycogen level but was lower than that obtained with group IX. From the study, it can be concluded that the combination of Pleurotus ostreatus with Murraya koenigii and Aegle marmelos has the highest hypoglycaemic and hypolipidaemic effect than the individual effect of Murraya koenigii and Aegle marmelos suggesting a synergistic action of the three.ThesisItem Open Access Hypoglycaemic effect of Coccinia indica (ivy gourd) leaves and its interaction with glibenclamide in diabetic rats(Department of Veterinary Pharmacology & Toxicology, COVAS, Mannuthy, 2009) Eliza Jose; KAU; Usha, P T AThe present study was undertaken to assess the hypoglycemic, hypolipidemic, antioxidant effect, interactive effects with glibenclamide and toxicity of ethanolic extract of Coccinia indica leaves. The experiment was conducted in seven groups of eight adult Sprague-Dawley rats each. Group I served as the normal control. All the groups except normal control were made diabetic by subcutaneous injection of alloxan at the dose rate of 130 mg/kg body weight and group II was kept as diabetic control. The groups III and IV were administered with the extract of C. indica at the dose rate of 200 mg/kg and glibenclamide at the dose rate of 0.25 mg/kg respectively for 45 days. The groups V, VI and VII were administered with a combination of C. indica at the dose rate of 100, 150 and 200 mg/kg respectively with glibenclamide at the dose rate of 0.125 mg/kg body weight for 45 days. Blood glucose, serum cholesterol and serum triglyceride and body weight were noted on day zero, 16th, 30th, 45th and 60th day and liver glycogen on 60th day. The antioxidant effect was assessed by estimation of reduced glutathione and lipid peroxides in pancreas and liver. Representative samples of pancreas were also subjected to histopathological examination. The diabetic control showed significant increase in the level of blood glucose, serum cholesterol and serum triglyceride level and also a significant reduction in body weight. All the other groups showed a gradual increase in body weight after 30 days of treatment. The groups III and IV restored the blood glucose levels after 45 days. The serum cholesterol, serum triglyceride, reduced glutathione and lipid peroxide levels in pancreas and liver were also restored indicating the hypolipidemic and antioxidant effect of the treatments. The groups V, VI and VII, in which the combinations of C. indica with glibenclamide were used, regained the blood glucose levels in 45 days of treatment. Group VII showed most marked effect and is found to be better than group III. The combination treatments produced a marked reduction in serum cholesterol and triglyceride levels and there was restoration of lipid peroxides and reduced glutathione in pancreas and liver. The ethanolic extract of C. indica leaves were screened for acute oral toxicity and sub acute toxicity. The administration of the extract upto 2000 mg/kg body weight (OECD guidelines 423) did not show any acute oral toxicity in mice. The sub acute toxicity studies conducted in wistar rats, did not show any toxic symptoms like mortality, change in biochemical, hematological parameters. The results revealed that C. indica did not possess any sub acute toxicity. These toxicity studies indicated the safety of ethanolic extract of C. indica as a drug in the treatment of diabetes. From the study it can be concluded that C. indica at the dose of 200 mg/kg body weight produced better hypoglycemic effect when compared with glibenclamide at the dose rate of 0.25 mg/kg body weight. Further the combination of C. indica ( 200 mg/kg ) with glibenclamide (0.125 mg/kg) showed a more effective hypoglycemic, hypolipidemic and antioxidant effect than C. indica or glibenclamide alone and the combination can be safely used in the treatment of diabetes.ThesisItem Open Access Hypoglycemic, hypolipidaemic and cytoprotective effects of lotus seeds (Nelumbo nucifera) in sprague-dawley rats(Department of Pharmacology and Toxicology, College of Veterinary and Animal Sciences, Mannuthy, 2007) Jeyamurugan, M; KAU; Chandrasekharan Nair, A MA study was undertaken to assess the hypoglycemic, hypolipidaemic and cytoprotective effects of ethanolic extract of red and white lotus seeds in male Sprague-Dawley rats. It was carried out in different animal models and compared with respective reference drug. The extracts of red and white lotus seeds were administered at the rate of 600 mg kg-1 for 28 days. The experiment was carried out in two hundred and seventy six rats. The blood glucose, serum total cholesterol, serum HDL, serum triglycerides and plasma free fatty acids was estimated on 0, 7th, 14th, 21st and 28th day of treatment in the respective animal models. The LDL cholesterol was calculated with help of formula. Type I diabetic rats treated with lotus seeds (red and white) extract showed significant reduction in blood glucose from 14th day onwards and in case of metformin from 7th day onwards. On 28th day oral glucose tolerance test and on 29th day liver glycogen and GLUT2 gene expressions were carried out. A significant increase in liver glycogen and reduction in GLUT2 gene expression was observed in all the treated groups. Glucose tolerance also showed significant improvement in all the treated group. In case of type II diabetic rats there was significant reduction in glucose levels from 21st day of treatment in extracts treated groups. Metformin treated group showed significant reduction from 7th day onwards. There was significant increase in liver glycogen, GLUT4 gene expression and glucose tolerance in all the treated groups. The pancreatectomized rats showed significant reduction in blood glucose levels from 21st day onwards in all the treated groups. Histopathological examination of pancreas revealed that there was significant proliferation of beta cells and increasing number of active cells was seen in all the treated groups. Hypercholesterolemic animals treated with lotus seed extract showed significant reduction in total cholesterol and LDL levels from 14th day treatment onwards. In case of simvastatin treated group significant reduction was noticed from 7th day treatment onwards. A significant increase in HDL level was observed on 28th day and from 7th day in extract and simvastatin treated groups respectively. A significant reduction in liver total lipid was noticed in all the treated group on 29th day. Hypertriglyceridemic animals treated with the extract showed significant reduction in triglycerides from 14th day treatment onwards. In case of fenofibrate significant reduction was observed from 7th day treatment onwards. The free fatty acid levels reduced significantly from 21st and 14th day onwards for extract and fenofibrate treated groups respectively. Liver total lipid level showed significant reduction in all the treated groups on 29th day. Pancreatitis was induced by repeated administration of alloxan in rats. They were treated with lotus seed extracts for 28 days. Pancreas was collected after sacrificing the animals for estimation of superoxide dismutase, catalase, and glutathione peroxidase and lipid peroxidation. The superoxide dismutase, catalase and glutathione peroxidase levels increased in pancreas of extract and metformin treated group. It was significant from 14th day treatment onwards. There was a significant reduction in lipid peroxidation level in red and white lotus seed extract treated group from 21st and 14th day treatment respectively. In case of metformin treated group significant reduction was noticed from 14th day of treatment. Histopathological examination of pancreas revealed islet hyperplasia with more number of active cells in a duration dependent fashion in all the treated groups. From the study, it can be concluded that both red and white lotus seed extract have hypoglycemic, hypolipidaemic and cytoprotective effects. However, the effects are less than that of respective reference drug.