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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, N
The 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.
Open Access
Antibacterial and antifungal activity of selected medicinl plants available in Kerala
(Department of Veterinary Pharmacology and Toxicology,College of Veterinary and Animal Sciences, Mannuthy, 2010) Sabitha, Jose; KAU; Usha, P T A
In the present study cold ethanolic extract and fresh juice of five medicinal plants were screened for their in vitro antibacterial and antifungal activities. The plants were Annona squamosa (Aatha), Cassia alata (Anathakara), Coleus amboinicus (Panicoorka), Myristica fragrans (Nutmeg) and Tectona grandis (Teak). Antimicrobial activity was tested against Staphylococcus aureus subsp.aureus (MTCC 96), Salmonella enteritidis (MTCC 3219), Escherichia coli (MTCC 723), Pasteurella multocida subsp.multocida (MTCC 1161), Pseudomonas aeruginosa (MTCC 741), Aspergillus fumigatus (MTCC 870), Candida albicans (MTCC 227) and Cryptococcus neoformans var neoformans (MTCC 4404). Phytochemical analysis was conducted for the presence of routine secondary plant metabolites. The diameter of inhibitory zone at various concentrations of the extract, minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and minimum fungicidal concentration (MFC) were used to evaluate the in vitro antimicrobial activity of the above mentioned plants. Disc diffusion method, microtitre plate dilution technique and broth dilution technique were used. The reference drugs used in this study were penicillin G, furazolidone, ketoconazole and clotrimazole. The maximum yield was obtained from ethanolic extract of A. squamosa leaves (18.07 per cent). Leaves of C. amboinicus produced the maximum amount of fresh juice among the five plants (8 ml from 10 g of the fresh tender leaves). Phytochemical analysis reported the presence of steroids, alkaloids, tannins, flavonoids, glycosides, phenolic compounds, diterpenes, triterpenes and saponins in the leaves of A. squamosa and C. alata. Alkaloids, tannins, flavonoids, glycosides, steroids and saponins were present in the leaves of C. amboinicus. M. fragrans leaves contain glycosides, flavonoids, saponins and phenolic compounds. Qualitative chemical tests revealed the presence of tannins, flavonoids, glycosides, phenolic compounds and saponins in the leaves of T. grandis. All the extracts showed varying degrees of antimicrobial activity on the microorganisms tested. The antimicrobial screening revealed the susceptibility of E. coli to A. squamosa, C. amboinicus and T. grandis. The growth of S. aureus, P. multocida and P. aeruginosa was inhibited by all the five plants. S. enteritidis was found to be susceptible to T. grandis, M. fragrans and C. amboinicus. MIC values ranged from 200-1000 µg/ml and MBC values ranged from 500-1000 µg/ml. In case of A. squamosa, MBC value against E. coli was more than 1000 µg/ml. The antifungal screening revealed that the growth of C. albicans was inhibited by C. alata, C. amboinicus, M. fragrans and T. grandis. A. squamosa and C. alata inhibited the growth of C. neoformans. A. fumigatus appeared to be susceptible to A. squamosa, C. alata and C. amboinicus. MIC values ranged from 250-1000 µg/ml for the fungal strains. MFC values ranged between 500-1000 µg/ml except for A. squamosa and C. alata. MFC of C. alata against A. fumigatus and C. albicans was more than 1000 µg/ml. MFC of A. squamosa against C. neoformans was found to be more than 1000 µg/ml. The growth of S. aureus was inhibited by fresh juice of A. squamosa, C. amboinicus, M. fragrans and T. grandis leaves. E. coli was susceptible to fresh juice of C. amboinicus leaves. Fresh juice of C. alata was found to be effective against C. albicans. P. aeruginosa was inhibited by fresh juice of M. fragrans leaves. All the plants under the study were found to possess antimicrobial properties, thereby justifying their popular use in the treatment of infectious diseases caused by resistant microorganisms. Further study is required to assess the in vivo efficacy of these plants for the said action.
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 A
The 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.
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 A
The 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.
Open Access
Antipyretic and CNS activity of seeds from red and white types of lotus (nelumbo nucifera) in albino rats
(Department of Pharmacology and Toxicology, College of Veterinary and Animal Sciences, Mannuthy, 2006) Deepa, P K; KAU; Usha, P T A
The antipyretic and CNS activity of the extract of the seeds of red and white Nelumbo nucifera was studied in albino rats. Qualitiative tests for the detection of phytochemicals showed the presence of steroids, alkaloids, flavonoids, glycosides, phenolic compounds, diterpenes, triterpenes and saponins. Pyrexia was induced by subcutaneous injection of 20% Brewer’s yeast suspension. Group I served as pyrexia control, group II was administered aspirin at the dose rate of 100 mg/kg body weight, Group III and IV with extract of red lotus seed and group V and VI with extract of white lotus seed at the dose rate of 400 and 600 mg/kg body weight respectively. The body temperature was recorded from zeroth to fifth hour at one hour interval. Haematological parameters and Serum ALT, AST levels were estimated at peak of pyrexia and five hours after drug administration. For CNS study group I was kept as normal control, Group II was administered chlorpromazine at the dose rate of 7 mg/kg body weight. Group III , IV V and VI were treated as in the case of antipyretic study. Actaphotometer and Rotarod were used to assess the spontaneous and forced motor activities respectively. Haematological parameters and serum ALT, AST levels were estimated at peak of tranquillization. The extract of white lotus seed at dose rate of 600 mg/kg body weight showed maximum antipyretic effect followed by red lotus at 600 mg/kg body weight and white lotus at the rate of 400 mg/kg body weight among the treatment groups. By the fifth hour the effect of white lotus seed extract at the dose rate of 600 mg/kg body weight was comparable to that of standard drug aspirin. Eventhough the extract of red lotus at the dose rate of 600 mg/kg body weight inhibited the spontaneous motor activity to the maximum initially, the extracts were found to be equipotent after 120 minutes of drug administration. But the activity was less when compared with the standard drug chlorpromazine. Comparison of the treatment groups showed that group IV showed maximum inhibition of forced motor activity followed by group VI and group III. The haematological and biochemical parameters assessed were within the normal range in both studies. From the present study it can be concluded that the extracts of red and while lotus seeds possessed potent antipyretic and CNS effects. The extract from the seeds of white lotus showed more potent CNS activity where as both the extracts were showing equipotent tranquillizing property.
Open Access
Anti-inflamatory and analgestic actions of red and white lotus seeds (nelumbo nucifera) in albino rats
(Department of Pharmacology and Toxicology, College of Veterinary and Animal Sciences, Mannuthy, 2006) Vikrama Chakravarthi, P; KAU; Gopakumar, N
Anti-inflammatory and analgesic effects of Nelumbo nucifera (lotus) seeds, red and white types were assessed and compared in albino rats. The carrageenin induced paw oedema was one of the acute inflammatory models followed in anti-inflammatory screening. The acute and chronic type pain models viz., tail flick method and experimental neuropathy model of analgesic activity, respectively, were adopted for analgesic screening. Diclofenac potassium at the rate of 3 mg/kg was used as a standard drug for both the studies. In cyclooxygenase-2 enzyme inhibition assays the celecoxib @ 10mg/kg was used as a standard drug. Both red and white types of lotus seeds at the dose rate of 400 mg/kg and 600 mg/kg were taken for the anti-inflammatory and analgesic studies. Anti-inflammatory effect of red and white lotus seeds was found to be effective in all phases of carrageenin induced inflammation. The higher dose groups of lotus seed extracts were revealed more inhibition than their corresponding lower dose. While comparing all groups, the higher dose group of white lotus seed, exhibited more pronounced inhibition of paw oedema than others. Analgesic effect was found to be significant in both acute and chronic analgesic models. The analgesic activity was more in tail flick method when compared to experimental neuropathy model. The cyclooxygenase-2 (COX-2) enzyme inhibition assays in both ELISA and spectrophotometer showed significant effect than control. The percentage of inhibition of COX-2 was more evidenced in both lotus seed extract of higher dose groups than its lower dose. However, the higher dose groups of white lotus seed exhibited more control over inhibition of COX-2 enzyme than others. . In case of biochemical parameters in both anti-inflammatory and analgesic screening the serum cholesterol level was found to be decreased in treatment groups when compared to control. Even though, there was a reduction in serum cholesterol level, all the values were within the normal range. The rise in serum level of AST and ALT in both inflammation and analgesic models were noticed in lower dose treated group. Conversely, there was a reduction in level of enzyme was noticed in higher dose treated groups. However, the values were with in the normal level. Both the studies showed the haematological parameters viz., total leukocyte and differential counts were within the normal range in all groups, even though a non-significant increase in neutrophil count than lymphocyte was noticed in carrageenin induced inflammation and tail flick method in rats.
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 M
A 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.
Open Access
Anti-ulcer effect of Azadirachta indica (neem) and Eupatorium triplinerve (Ayyappana) in rats
(Department of Pharmacology and Toxicology, College of Veterinary and Animal Sciences, Mannuthy, 2005) Sangeetha Satheesan; KAU; Joy, A D
The present study was conducted in adult albino rats to assess the anti-ulcer effect of alcoholic extract and powder of leaves of Azadirachta indica and Eupatorium triplinerve in comparison with famotidine, a standard anti-ulcer drug. Ninety six rats weighing 150-200gm body weight of either sex divided into twelve groups were used for the study with eight rats in each group. Group 1 was administered the vehicle, five per cent gum acacia for seven days where as group 2 was administered aspirin at the dose of 200mg/kg for seven days. Group 3 was administered aspirin for seven days and from 8th day onwards, they were maintained by normal feeding and watering for 20 days to assess the natural healing. Famotidine, a standard anti-ulcer drug was given at the dose of 40mg/kg for 20 days following aspirin administration to the group 4 animals. The alcoholic extract and powder of A.indica and E.triplinerve were administered to the treatment groups (Groups 5-12) at the two dose levels of 250mg/kg, 500mg/kg and 500mg/kg, 1000mg/kg respectively for 20 days following aspirin administration. The control group as well as the aspirin group were sacrificed on the 8th day, whereas all the other groups were sacrificed on 28th day. The number of ulcers and severity (ulcer score) were determined with the help of magnifying lens and the ulcer index and healing index were calculated. Various biochemical parameters were studied to confirm the anti-ulcer activity of the plant preparations under study. The degree of lipid peroxidation, as well as the anti-oxidant enzyme status, namely, superoxide dismutase and catalase levels were assessed in gastric mucosa. The serum alkaline phosphatase activity was estimated on 28th day of the experiment. Body weight was taken at weekly intervals. Heamatological parameters such as total leucocyte count, differential count and haemoglobin count were determined to assess any changes in the haemogram. Histopathological study was also conducted to evaluate the severity of ulceration and healing process. The results indicated that all the treatment groups under study produced a significant decrease in ulcer index when compared to aspirin treated control group. The alcoholic extract at the two dose levels 250mg/kg and 500mg/kg as well as powder at the two dose levels 500mg/kg and 1000mg/kg of both A.indica and E.triplinerve were found to produce a dose-dependent healing effect which is comparable to that of famotidine. Administration of the herbal formulation at the various dose levels brought about a significant reduction in lipid peroxidation and an increase in the activities of anti-oxidant enzymes namely, superoxide dismutase and catalase, which suggest its efficacy in preventing free radical induced damage. The treatment reversed the increased activity of serum alkaline phosphatase observed in the aspirin treated group. There was no significant difference in the mean body weight gain between any of the groups. Haematological study revealed no significant change and all values fall within the normal range of blood value for the species under study. The results are substantiated by the histopathological studies, which confirmed that treatment with A.indica and E.triplinerve inhibited aspirin-induced necrosis, haemorrhage and congestion in gastric mucosa. Phytochemical analysis of the extracts revealed the presence of tannins, saponins, flavonoids, triterpenes, alkaloids, phenolic compounds which are known to affect the integrity of mucous membranes. In the present study, the fact that A.indica and E.triplinerve significantly reduced ulcer index in a dose-dependent manner supports their cytoprotective effect, which may be mediated by prostaglandins and the ulcer healing effect could be attributed to its predominant effect on the mucosal defensive factors rather than offensive factors.
Open Access
Immunomodulatory effect of fractions of ethanolic extract from Emblica officinalis (Amla) fruit pulp in mice
(Department of Pharmacology and Toxicology, College of Veterinary and Animal Sciences, Mannuthy, 2005) Senthil Kumar, P K; KAU; Chandrasekharan Nair, A M
The immunomodulatory activity of acetone soluble and acetone insoluble fractions of ethanolic extract of Emblica officinalis was investigated in the present study. The extracts were also qualitatively tested for the presence of various active principles. One hundred and forty four male swiss albino mice were used to assess humoral and cellular immunity by feeding the extracts at dose level of 200 mg per kg body weight for 19 days. Few animals were also administered with dexamethasone at the dose rate of 0.75 mg per kg body weight intra peritoneally for seven days before the start of the experiment to suppress the immune system and thereby to study the effect of extracts on immunosuppressed animals also. The control group received vehicle alone (five percent gum acacia). Various physiological, haematological, biochemical, enzymatic and immunological parameters like body weight, relative organ weight, total leukocyte count, differential leukocyte count, total serum, protein, serum globulin, albumin-globulin ratio, quantification of superoxide dismutase and catalase, haemagglutination test, Jerene’s plaque forming assay to assess the humoral immune response and tests like delayed type of hypersensitivity, macrophage migration index, nitroblue tetrazolium reduction test to assess the cellular immune response were performed. Both the fractions increased the body weight, spleen weight and liver weight in normal as well as in immunosuppressed animals. A significantly increased total WBC counts on days 5, 12 and 19 and the distribution of lymphocytes on days 12 and 19 were seen in normal animals. In immunosuppressed animals, non-significant increase in both total leukocyte counts and distribution of lymphocytes from day zero to nineteen was seen. Total serum protein as well as serum globulin concentration was significantly increased and albumin globulin ratio was significantly decreased in immunocompetent animals treated with both the fractions on 12th and 19th day. However, both the fractions showed only a non-significant increase in total serum protein and serum globulin in normal as well as in immunosuppressed animals, from zero day to 19th day. Significant increase was noticed in superoxide dismutase and catalase level in immunocompetent animals treated with both the fractions on 19th day. A significant stimulation of humoral immune response as indicated by an increase in antibody titre and number of antibody producing cells on both 12th and 19th day of acetone soluble and acetone insoluble fraction treated immunocompetent animals was noted. Immunosuppressed animals also showed a non-significant increase in both antibody titre and number of antibody producing cells through out the experiment. The bone marrow cellularity and foot pad swelling reaction showed a significant stimulation in immunocompetent animals on 12th and 19th day. In immunosuppressed animals also, foot pad swelling response showed a significant increase on 19th day but bone marrow cellularity was not significant. In immunocompetent animals the acetone soluble and acetone insoluble fractions showed a significant increase in macrophage migration index (MMI) and Nitro blue Tetrazolium (NBT) dye reduction test value on 12th and 19th day. However, in immunosuppressed animals both the fractions showed only a non-significant increase. The phytochemical study on both the fractions of ethanolic extract of dried Emblica officinalis fruit pulp revealed that diterpenes and triterpenes were present in acetone soluble fraction and tannins, phenolic compounds, flavonoids, glycosides, and saponins were present in acetone insoluble fraction. Thus the present study showed the higher immunostimulant activity for acetone soluble fractions in immunocompetent as well as immunosuppressed animals.

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