Evaluvation of Analgesic, Anti-Inflammatory and Cytotoxic Studies of the Methanolic Extract of Sphenodesme Paniculata (Clarke) in Experimental Animals

INTRODUCTION: Medicinal herbs are significant source of semisynthetic and herbal formulations as in commercial market for various illness. Isolated active constituents are used for applied research. The plant Sphenodesme paniculata is locally known as Arambodal / Njarambodal. Ethno botanically, the plants are used by the tribals of pathanamthitta district, kerala, for the treatment of body pain. This plant also used as anti-inflammatory and wound healing agent in part of traditional medicinal formulations that alleviate body pains.11The plant Sphenodesme paniculatais used in the present study to confirm the biological properties by invivo screening method. AIM: Sphenodesm epaniculata (Clarke) isa Climbing or scandent shrubs, with traditionally medicinal plants used for treating wound and body pain by the tribals in Pathanamthitta district, Kerala, India. Current literature study revealed that the anti-inflammatory and analgesics activity of Sphenodesmepaniculata (Clarke) have not being studied. Hence the aim of the present study is to evaluate analgesics, anti-inflammatory and cytotoxicity studies of methanolic stem extract of Sphenodesme paniculata (Clarke). OBJECTIVES: 1. Collection of the plant, 2. Authentification, 3. Extraction of stem, 4. Phytochemical analysis, 5. Qualitative analysis, 6. Quantitative analysis, 7. In-vitro antioxidant studies, 8. DPPH, 9. ABTS, 10. Cytotoxicity activity, 11. HeLa cell line, 12. Anti microbial activity, 13. Disc diffusion method, 14. Invivo studies, 15 Acute toxicity study, 16. Evaluation of the analgesic, anti inflammatory activities of the methanolic stem extract of Sphenodesme paniculata. SUMMARY AND CONCLUSIONS: I. Preparation of 95% methanolic extracts, ethyl acetate extracts, of sphenodesme paniculata stemby using Soxhlet extraction andthe aqueous extraction was carried out by boiling the coarsely powdered leaves with waterbased on the polarity. II. All the extracts were subjected to various phytochemical screening and the study reveals the presence of alkaloids, phenols, flavanoids, carbohydrate ,steroids, saponins and glycosides were found in the stem extracts of sphenodesme paniculata. III. Total phenol content (TPC) for the extracts of sphenodesme paniculata were determined by Folin-Ciocalteu’s method and it was found that the methanolic extract contains high amount of total phenols which was about 68.6±0.02 mgGAE/g for the MESPand 41.4±0.02 mg GAE/g for the EESP. IV. Total flavanoids content of all the extracts of sphenodesme paniculata were determined by aluminium chloride colorimetric assay. It was found found that the extracts contains high amount of total flavanoids which are about 28.47±0.01 mg QE/g for the MESP and 21.1±0.02 mg QE/g for the EESP. V. Total antioxidant activity of theMESP extracts was performed.Total antioxidant capacity of MESP was found to be 342.8 μg/ml calculated as Ascorbic acid equivalent. VI. In the present study, Simultaneously determinationof quercetin, rutin and gallic acid in sphenodesme paniculatastem is carried out by HPTLC in solvent system of toluene : ethyl acetate: formic acid : methanol (3:0.8:0.2). VII. The MESP extracts were evaluated for its antioxidant potential. DPPH radical scavenging ability of the extracts was studied. The IC50 values were found to be 6.4 μg/ml, extract showed potent anti oxidant activity. VIII. ABTS radical scavenging ability of the MESP extracts were studied. The IC50 values were found to be 5.97 μg/ml, extract showed potent anti oxidant activity. IX. The cytotoxicity of all the stem extracts of sphenodesme paniculata were performed by MTT assay using HeLacell lines. The cytotoxicity experiment showed that all tested extracts were non-cytotoxic to HeLa cells up to a concentration of 200 μg/mlX. Antimicrobial activity of aii the stem extract of sphenodesme paniculata were performed by sensitivity study, all the extracts showed antimicrobial activity, methanolic extract showed higher zone of inhibition compare to other extracts XI. The MESP extracts wereused for the pharmacological evaluvation of Anti-inflammatory activity at the doses of 200 and 400 mg/kg, methanolic extract showed significant reduction in paw oedema in carrageenan and histamine induced model when compared with control to treated group. XII. The MESP extracts were used for the screening of thermal and chemical induced nociception (Hot plate and Writhing response) at the doses of 200 and 400 mg/kg, methanolic extract showed significantly reduced thermal and chemical induced nociception when compared with control to treated group. The obtained results from overall study supports that the stem extracts of sphenodesme paniculata having potent antioxidant activity, andante-microbial activity, The cytotoxicity studies on HeLa cell line proved that the extracts were not having any cytotoxic effect. plant exract showing significant activity in analgesics and anti inflamatory. It is concluded that stem of this plant have medicinal important and beneficial for humans