ANTI-INFLAMMATORY ACTIVITY OF WRIGHTIA TINCTORIA LEAVES BY MEMBRANE STABILIZATION Rajalakshmi G.R. 1* , Jyoti Harindran 2 1. Research Scholar, Karpagam University, Karpagam, Coimbatore, Tamil Nadu-641021, India. 2. Principal, University College of Pharmaceutical Sciences, Mahatma Gandhi University, Rubber Board (P.O), Kottayam , Kerala, India. E mail: sijuellickal@rediffmail.com ABSTRACT Ethyl alcohol and aqueous extract of Wrightia tinctoria were investigated for anti-inflammatory activity by HRBC method. The preventation of hypo tonicity induced HRBC membrane lysis was taken a measure of anti- inflammatory activity and these extracts shows biphasic effects. Their activities are compared with standard drug diclofinac sodium. Key Words: Wrightia tinctoria, anti-inflammatory, Phytoconstituents. INTRODUCTION The inflammatory process is the responses to an injurious stimulus. It can be evoked by a wide variety of noxious agents. The ability to mount an inflammatory response is essential for survival in the face of environmental pathogens and injury; in some situations and diseases, the inflammatory responses may be exaggerated and sustained without apparent benefit and even with severe adverse consequences [1]. So present study was undertaken to establish scientific evidence for anti-inflammatory activity of leaves extracts of Wrightia tinctoria. Wrightia tinctoria is a member of the family Apocynaceae, is a small to medium-size deciduous tree [2]. Traditionally Wrightia tinctoria commonly called as “Jaundice curative tree” in south India and plant possesses high medicinal value [3]. Crushed fresh leaves when filled in the cavity of decayed tooth relieve toothache. In Siddha system of medicine, it is used for psoriasis and other skin diseases [4-6]. The plant has been assigned to analgesic, anti-inflammatory and antipyretic activities and to be effective in the treatment of psoriasis [7-8]. The literature survey revealed that no reports were found on the anti-inflammatory activity of the leaves extracts of Wrightia tinctoria. This prompted us to investigate the anti-inflammatory activity of Wrightia tinctoria leaves extracts. MATERIALS AND METHODS Plant Material Fresh leaves of Wrightia tinctoria was collected from TBGRI, Thiruvananthapuram during the month of March 2007. The plant was identified by Mrs. Amina Ali, Associate Professor and Head, Department of Pharmacognosy, Govt. Medical College, Calicut, Kerala, India. Voucher specimen (AA-34/10) is preserved in institute herbarium for future reference. Preparation of Extract Ethyl alcohol extract: The shade dried powdered leaves (500g) were exhaustively extracted with 95% ethanol using a soxhlet apparatus. The extract was concentrated in vaccuo to a syrupy consistency. The percentage yield of extract was found to be 2.9 %. Aqueous extract: The dried powders (24#) 100gm of the was taken in a 2000ml conical flask with 500ml of distilled water to which 10ml chloroform were added as a preservative. It was extracted up to 7 days with daily 2 hours stirring with the mechanical stirrer. After 7 days the extract was filtered through the muslin cloth and the marc was pressed and its filtrate dried in hot air oven at 45 0 C to a semisolid mass. It was stored in airtight container in a refrigerator below 10 0 C. The percentage yield of extract was found to be 3.1 %. Membrane stabilization assay The HRBC membrane stabilization has been used as method to study the anti-inflammatory activity [9] (Gandhisan etal. 1991). Blood was collected from healthy volunteer who was not taken any NSAIDS for two weeks prior to the experiment. The collected blood was mixed with equal volume of sterilized Alsever solution (2%dextrose,0.8% sodium citrate,0.5% citric acid and 0.42% sodium chloride in water). The blood was centrifuged at 3000 rpm and packed cell were washed with isosaline (0.85%.pH 7.2) and a 10 %( v/v) suspension was made with isosaline. The assay mixture contained the drug (concentration as mentioned in the table 2), 1 ml of phosphate buffer (0.15M, pH 7.4), 2 ml of hyposaline (0.36%) and 0.5ml of HRBC suspension. Rajalakshmi G.R. et al. / International Journal of Pharma Sciences and Research (IJPSR) ISSN : 0975-9492 Vol 3 No 10 Oct 2012 497