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118 Abstract Evaluation of In-Vivo Anti-Rheumatic EVALUATION OF IN-VIVO ANTI-RHEUMATIC ACTIVITY OF ANISOMELES MALABARICA R.BR. Ismail Shareef. M1, Leelavathi. S2, Thara Saraswathi. K. J3, Sampath Kumara K. K4 1 Department of Biotechnology, Acharya Institute of Technology, Soladevanahalli, Bangalore, Karnataka 2 ijcrr Manasagangotri, University of Mysore, Mysore, Karnataka Vol 04 issue 13 3Department of Microbiology & Biotechnology, Jnana Bharathi Campus, Bangalore Category: Research University, Bangalore, Karnataka Received on:28/05/12 4Government P.U College for Boys, Davengere, Karnataka Revised on:05/06/12 E-mail of Corresponding Author: [email protected] Accepted on:11/06/12 ABSTRACT The present study was ca rried out to investigate the anti-rheumatic properties of methanolic extracts of the aerial parts of the plant Anisomeles malabarica R.Br., (Lamiaceae) using experimental animal models. The anti-rheumatic activity of the methanolic extracts was studied based on the effects on carrageenan-induced rat paw oedema. The preliminary phytochemicals were screened for the presence and absence of alkaloids, steroids, proteins, flavonoids, saponins, carbohydrates, tannins, fats and oils. The extracts in dose level 2000 mg/kg orally were used in anti-rheumatic studies. The methanolic extracts of leaves of Anisomeles malabarica R.Br., produced significant anti-rheumatic activity in a dose- dependent manner (200 mg/Kg and 400 mg/Kg body weight) to that of standard drug indomethacin (10 mg/Kg). The extract exhibited inhibitory effect in carrageenan induced hind paw oedema in rats with all the doses used when compared to the control group. The data obtained indicate that the crude extracts of the aerial parts of the plant Anisomeles malabarica R.Br., possess potential anti-rheumatic activity by supporting the folkloric usage of the plant to treat various inflammatory conditions. Keywords: Anisomeles malabarica R.Br., Anti-rheumatic, Carrageenan, Rat paw oedema ____________________________________________________________________________________ INTRODUCTION untreated. This disease has a worldwide Inflammation is a normal protective response to distribution but its pathogenesis is not clearly tissue injury that is caused by physical trauma, understood2 although there are few anti-rheumatic noxious chemicals or microbiological agents. drugs showing effectiveness in the treatment of Inflammation is the result of concerted rheumatoid arthritis, the side effect and toxicity participation of a large number of vasoactive, call for new and more effective natural drugs3. chemotactic and proliferative factors at different There are many herbs which have been enlisted to stages and there are many targets for anti have the potential in the symptomatic treatment of inflammatory action1 . rheumatoid arthritis, hence the present study has Rheumatoid arthritis is a systemic autoimmune been carried out to evaluate the anti-rheumatic disorder characterized by polyarticular potential. The tools of biotechnology provides vast symmetrical arthritis. Various inflammatory potential for the development of new inventions, mediators produce joint inflammation with pain particularly in the field of pharmaceuticals which function loss, joint destruction and permanent are environmentally safe and do not require heavy deformity after certain time if remained 118 International Journal of Current Research and Review www.ijcrr.com Vol. 04 issue 13 July 2012 investments. In this context, the herbal medicines was poured into the flask (distilling pot) and the have been proved to have tremendous scope4. whole apparatus was set. The soxhlet extraction Anisomeles malabarica R.Br. (Lamiaceae) is was performed for 12- 16 hours until the collected distributed in major parts of India and especially in solvent in siphon tube appears to be clear. Later South India as a traditional medicinal plant the extracted solvent was evaporated under commonly known as Peymarutti (Tamil), reduced pressure to get solid/ semi solid extract. Gouzaban (Hindi), Chodhara (Marathi), The extract was weighed, physical characters were Karithumbi (Kannada) and Malabar catmint noted. The percentage yield was calculated to be (English)5. The herb is reported to possess anti- 10.62. spasmodic, anti-periodic properties and used in Phytochemical screening rheumatoid arthritis6. It is used for the traditional All the extracts were screened for the presence of treatment of snakebite as antidote7 and plant leaves various active plant metabolites like steroids, are used as carminative, astringent, stomachic, alkaloids, carbohydrates, flavanoids, glycosides rheumatism and diaphoretic in Coimbatore and tannins according to standard phytochemical district8 and also used as dentifrice to cure various methods12. Briefly, Dragendorff reaction was used problems9. Preliminary phytochemical tests were to confirm the presence of alkaloids, alkaline carried out by Brindha et al., (1977)10 and used reagent test for tannins, frothing test for saponins, for the treatment of various infections. legal‘s test for glycosides, Xanthoproteic test for proteins and Shinoda test for flavanoids MATERIALS AND METHODS respectively. Chemicals 1. Acute toxicity study in rats with test drugs13, 14 Extraction was carried out using methanol in Two groups, each of three female rats, were soxhlet apparatus. Chemicals & reagents for the treated with the extracts of the aerial parts of the present analysis were purchased from Karnataka plant namely Anisomeles malabarica (abbreviated fine chemicals, Bangalore and E. Merck Ltd., as AmA/Test drug) by Oral administration at a Mumbai, India. dosage of 2000 mg/kg body weight. The test drug Plant material was formulated in vehicle (distilled water) at a Anisomeles malabarica was collected from concentration of 2000 mg/mL and administered at Mysore, Nanjangud and surrounding areas and the dose volume of 10 mL/kg. The animals were also from medicinal garden of Indian Institute of observed daily during the acclimatization period Horticultural Research, Hesaraghatta, Bangalore and mortality/viability and clinical signs were where they were growing profusely. The plants recorded. All animals were observed for clinical have been identified and authenticated by experts signs during first 30 minutes and at approximately from National Ayurveda Dietetics Research 1, 2, 3 and 4 hours after administration on test day Institute, Bangalore (Ref. No. 0 and once daily during test days 1-14. SMPU/NADRI/BNG/2010-11/550). Mortality/viability was recorded twice daily Extraction of plant material during days 1-14 (at least once on day of The plant materials were extracted with methanol sacrifice). Body weights were recorded on test using sohxlet extraction apparatus continuously day 0 (prior to administration), test days 7 and 14 for 16 hours11. For extraction, the dried plant (Table 1). All animals were necropsied and material was used. Initially 400gms of material examined macroscopically (Table 2). was packed in filter paper and loaded into the thimble of soxhlet apparatus. 2.5 liter of methanol 119 International Journal of Current Research and Review www.ijcrr.com Vol. 04 issue 13 July 2012 Treatment 14 h light dark cycle. They were provided with The animals received a single dose of the test item Amrut brand balanced feed and tap water ad by oral administration at 2000 mg/kg body weight libitum. 16 after being fasted for approximately 18.0 hours but 3. Carrageenan induced rat paw oedema with free access to water. Food was provided Thirty rats were divided into five groups (n=6) again at approximately 3.0 hours after dosing. The starved overnight with water ad libitum prior to administration volume was 10 mL/kg body the day of experiment. The group I kept as control weight. The animals were dosed using 18 G oral group, group II kept as carrageenan control, Stainless steel feeding tubes. groups III and IV received test drug at different Necroscopy doses and group V kept as standard drug control, All animals were sacrificed at the end of the respectively. Left paw was marked with ink at the observation period by carbon dioxide in euthanasia level of lateral malleolus; basal paw volume was chamber and discarded after the measured plethysmographically by volume gross/macroscopic pathological changes were displacement method using Plethysmometer (UGO observed and recorded. No organs or tissues were Basile 7140) by immersing the paw till the level of retained. lateral malleolus. 2. Anti-inflammatory studies15 In the experiment, animals from the control group This model is based on the principle of release of were given vehicle control (CMC) and animals various inflammatory mediators by carrageenan. from standard drug were treated with Oedema formation due to carrageenan in the rat Indomethacin as shown in Table 3. Other groups paw is biphasic event. The initial phase is were treated with different doses of test drugs as attributed to the release of histamine and shown in Table 3. After 30 min. of drug treatment serotonin. The second phase of oedema is due to the rats are challenged by a subcutaneous injection the release of prostaglandins, protease and of 0.1ml of 1% solution of carrageenan into the lysosome. Subcutaneous injection of carrageenan sub-plantar side of the left hind paw. The paw into the rat paw produces inflammation resulting volume is measured again at 1, 2, 3, 4 & 5 hours from plasma extravasations, increased tissue water after challenge. The increase in paw volume is and plasma protein exudation along with calculated as percentage compared
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