PHYTOCHEMICAL COMPOSITION, ANTIOXIDANT AND IN-VITRO ANTI-INFLAMMATORY ACTIVITY OF ETHANOL EXTRACT OF RUTA GRAVEOLENS L. LEAVES Original Article GIRESHA A. S., ANITHA M. G., DHARMAPPA K. K. * Department of studies in Biochemistry, Mangalore University, P. G centre, Chikka aluvara, Kodagu, India Email: dharmappa@gmail.com Received: 14 Jan 2015 Revised and Accepted: 20 Aug 2015 ABSTRACT Objective: The uses of medicinal herbs in prevention and treatment of numerous diseases attracting attention of scientists worldwide. It is necessary to bring vital healthcare to the people and the better substitute for various infectious diseases. Ruta graveolens is traditionally used as a medicinal plant as well as a flavoring agent in food. In the present study R. graveolens leaves extracts were evaluated for phytochemical and anti- inflammatory activity. Methods: The phytochemical, antioxidant (DPPH), ferrous reducing antioxidant property, Anti-lipid peroxidation and superoxide scavenging activity of extracts were assayed by spectrophotometerically. Anti-inflammatory activities; Human plural fluid phospholipase A2 (HPF-PLA2) inhibition by egg yolk plate method and lipoxigenase (LOX) inhibition were assayed by IN VITRO method. Results: The phenolics were significantly higher than the flavonoids, saponins and alkaloids contents in all the extracts. Among extracts of R. graveolens, ethanol and water extracts exhibits more phytochemicals. The ethanol extract had the highest antioxidant activity followed by the aqueous extract. The ethanol extract shows highest DPPH free radical scavenging, ferric reducing, superoxide scavenging and anti-lipid peroxidation activity and IC50 was 3.27±0.03, 3.58±0.05, 3.87±0.04 and 4.77±0.04 µg/µl respectively at dose-dependent manner. Further ethanol and water extracts were subjected for LOX inhibition in the concentration dependent manner and IC50 values are 4.25±0.05, 5.15±0.05 µg/µl for 5-LOX and 4.15±0.04, 4.66±0.05 µg/µl for 15-LOX respectively. Conclusion: The result shows R. graveolens has strong antioxidant property and anti-inflammatory activity. It suggests that the strong correlation between antioxidant activity and the phytochemical contents of the extracts. Keywords: HPF, Human Pleural Fluid, Anti-inflammatory, sPLA2, Secretory Phospholipase A2, HPF-PLA2, Human Pleural Fluid-PLA2. LOX Lipoxigenase, Lipid peroxidation, Anti-inflammation. INTRODUCTION Plants are the local heritage with global importance. World is endowed with a rich wealth of medicinal plants. Plant extracts have been the source of remedy for various clinical abnormalities from the time immemorial. Over three quarters of the world population relies mainly on plant-based medicines for the primary health care [1]. Roughly more than 30% of the plant species have been used for therapeutic purposes and about 25% of all prescribed medicines using today are substances derived from plants. Many of these indigenous medicinal plants are used as spices and food plants [2]. They are sometimes added to foods meant for pregnant and nursing mothers for medicinal purposes [3]. The medicinal properties of herbal extracts are attributed to the presence of diverse classes of bio-active molecules, many of which have attained the level of drugs [4]. Phytochemicals are employed for the treatment of hypertension, cardiovascular disorders, cancer, neuronal disorders, inflammation, diabetes, malaria, influenza, arthritis, and severe pain [5]. Inflammation is a localized reaction that produces redness, warmth, swelling, and pain result in infection, irritation or injury. It contributes to all disease process including immunity, vascular pathology and leads to several undesirable consequences such as systemic shock, circulatory collapse and local tissue injury in many organs [6, 7]. Inflammation is mainly mediated by secretory phospholipase A2 (sPLA2). Several snake venom PLA2 enzymes are responsible for local tissue damage such as edema and hemorrhage. Elevated level of sPLA2 enzymes are detected in many inflammatory disorders [8, 9] and play a key role by releasing arachidonic acid and lysophospholipid, which are rate-limiting precursors for the production of pro-inflammatory lipid mediators. Cyclooxigenase-1/2 (COX-1/2) and lipoxigenase (LOX) catalyze arachidonic acid into pro-inflammatory mediators such as prostoglandins, thrombaxanes, and leukotriens respectively. Lysophospholipid is further converted into platelet activation factor (PAF) by acetyltransferase [10]. Further, Phospholipase A2 (PLA2) plays a crucial role in a number of diverse cellular as well as chemical and metabolic injuries. Apart, they involved in phospholipid digestion and metabolism, host defense, signal transduction and provide precursors for eicosanoid generation. Present anti-inflammatory therapies include the non-steroidal anti- inflammatory drugs (NSAIDs) that inhibit either Cyclo-oxygenase- 1/2 (COX-1/2) or Lipoxygenase (LOX) catalysis have severe side effects such as intestinal ulceration, bleeding and cardiovascular complications [11]. In addition to these problems, COX-1/2 or LOX inhibitors cannot regulate the production of leukotrienes or PAF that continue to cause inflammation [12]. It appears rational that effective inhibitors of sPLA2 could deplete the sources of arachidonic acid and therefore its downstream metabolites and PAF would not affect the homeostasis of COX-1/2 and LOX enzymes. Therefore, the great demand for natural products for PLA2 inhibition rather than NSAIDs to treat inflammatory disorders. The literature survey showed that several endogenous and exogenous agents have been reported to inhibit sPLA2 enzymes [13]. In addition, several laboratories are synthesizing compounds to inhibit sPLA2 [14]. But none of them are made into the market, which demands researchers to look for new specific sPLA2 inhibitors. Ruta graveolens belonging to family Rutaceae has long been used traditional medicine for various ailments including hysteria, gastrointestinal disorders, and menstrual problems. The juice of fresh rue has been used to relieve toothaches and earaches. It has been reported to possess antifungal [15], antibacterial [16], anti-inflammatory [17], antitumour [18] and cytotoxic activities [19]. Although, no information available on the effect of R. graveolens on inflammatory enzymes. In the present study, effect of R. graveolens leaves ethanol and aqueous extract on LOX and sPLA2 activity was estimated. MATERIALS AND METHODS Diphenyl Picryl Hydrazyl radical (DPPH˙), Thiobarbituric acid, Quercetin, Gallic acid, were purchased from Sigma-Aldrich Chemical International Journal of Pharmacy and Pharmaceutical Sciences ISSN- 0975-1491 Vol 7, Issue 10, 2015 Innovare Academic Sciences