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ThesisItem 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 DThe 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.ThesisItem 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 MThe 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.