Soubhagya K B

Evaluation of Anti-oxidant and Anti-inflammatory Activity in Leaf and Flower Ethanolic Extracts of Nyctanthes arbor-tristis Linn. Soubhagya K. 1* 1*, a) B. and Dr. M. 2,b) Anilkumar Mar Athanasius College(Autonomous), Kothamangalam India. 2 Union Christian College, Aluva, India. ABSTRACT RESULT Studies on natural compounds are of interest in now a day, due to their satisfactory health benefits with low toxicity. Herbs and spices have been added to different types of food to impart flavour as well as to improve storage stability due to their anti-oxidant effects. Anti-oxidants play a major role in curing degenerative diseases. In current scenario many synthetic antioxidants are commonly used but due to their toxic and carcinogenic effects their use has been restricted by legislative rules. So the search for natural anti-oxidants is essential. In addition to the body's response to injury, inflammation is a physiological process underlying the progression various diseases. Investigation for the discovery and development of anti-inflammatory agents from natural sources is a promising strategy for curing inflammatory diseases. Leukocytes are the key players of inflammatory response can eliminate microbes and dead cells by phagocytosis, followed by their destruction in phagolysosomes. The destruction is due to the free radicals generated in activated leukocytes and lysosomal enzymes. The human RBC (HRBC) membranes are similar to these lysosomal membrane components and most anti-inflammatory drugs are targeted on these pathways. Preventing the release of inflammatory mediators is a better option than blocking a particular mediator or its pathway. Usually, conventional anti-inflammatory drugs are used to treat inflammations but they have side effects. Traditionally, Nyctanthes arbor-tristis Linn. (Nyctanthaceae) is used against a wide range of diseases in Indian traditional medicine. Phytochemical analysis of Nyctanthes arbor-tristis Linn. revealed the presence of biochemical compounds such as terpenes, steroids, tannins, phenols, reducing sugars, flavonoids, proteins etc.. Since triterpenoids and and flavonoids have remarkable antiinflammatory activity. Here, the ability of N. arbor-tristis Linn. ethanolic leaf and flower extracts is studied by HRBC membrane stabilization. The inhibition of hypotonicity induced HRBC membrane lysis was taken as a measure of anti inflammatory activity. Diclofenac was used as reference standard. The percentage of membrane stabilization of ethanolic leaf extract, flower extract and Diclofenac were estimated spectrophotometrically at different concentrations such as 200 µL, 400 µL, 600 µL, 800 µL and 1000 µL. They were found to be effective in protecting HRBC membrane. Leaf and flower extracts has a percentage of stabilization less than that of the reference standard, Diclofenac. Among the plant extracts, leaf shows higher stabilization than that of flower. OBJECTIVES The present study in N. arbor-tristis Linn. has been undertaken with the following objectives: • Evaluation of anti-oxidant activity by ferric reducing power ability of ethanolic extracts of leaf and flower. • Anti-inflammatory study by membrane stabilisation method using ethanolic extracts of leaf and flower. METHODOLOGY Determination of in-vitro Anti-oxidant activity - Ferric Reducing Ability Power The ethanol extracts of leaves and flowers of N. arbor-tristis Linn. was determined for their reducing power modifying the method of 1. Add 1 ml of phosphate buffer (0.2 M, pH 6.6), 1 ml Potassium ferric cyanide (1%) and varying concentrations of extracts separately (100 – 500 µl) The reaction mixtures were incubated at 500C in water bath for 30 minutes, allowed to cool at room temperature (280), and 2.5 ml of 1% TCA (trichloroacetic acid) were added to each reaction mixture, and then centrifuged at 2000 rpm for 10 minutes. The supernatants (2.5 ml) were separated in the test tubes and added with 3 ml of distilled water and 0.5 ml ferric chloride (1%), and then allowed to react for 10 minutes at room temperature and the absorbance was measured at 700 nm. Ascorbic acid solution (10%) was used as standard. Determination of in-vitro Anti-inflammatory activity Anti inflammatory activity of the ethanolic extracts of leaf and flower of Nyctanthes was carried out as described by2 . The blood was collected in heparinised tubes. The blood samples were centrifuged at 3000 rpm for 10 min at room temperature. The supernatants (plasma and leukocytes) were carefully removed while the packed RBC was washed in fresh normal saline (0.85% w/v NaCl). The process of washing and centrifugation were repeated five times until clear supernatants were obtained. From this a 1% (v/v) human erythrocyte suspension was prepared using isosaline. Diclofenac was used as standard drug. The assay mixtures consisted of 2 ml of hyposaline (0.25% w/v) NaCl, 1.0 ml of 0.15 M sodium phosphate buffer at pH 7.4 (25oC), 0.5 ml of 10% (v/v) HRBC suspension, 0.2-1.0 ml of drug/extract ( concentrations ranging from 400µg - 2000µg) and final reaction mixtures were made up to 4.5 ml with isosaline. Drugs were omitted in the blood control, while the drug control did not contain the erythrocyte suspension. The reaction mixtures were incubated at 40oC for 45 min on a water bath, followed by centrifugation at 3000rpm for 10 min at room temperature. The absorbance of the lysed haemoglobin was read at 560 nm using UV-Vis Spectrophotometer (shimadzu, Japan). RESEARCH POSTER PRESENTATION DESIGN © 2019 www.PosterPresentations.com CONCLUSIONS Nyctanthes arbortristis Linn. is easily available plant and no special conditions are required for its collection and cultivation. It is a rich source of biologically active compounds, which would attract the attention of drug discovery groups to discover novel bioactive molecules for safer and effective treatment of various diseases. Though plant has been found to be safe but the potent curative effects of the plant against particular human ailments need to be verified by more controlled and exhaustive clinical trials. In the present study, Ferric reducing power ability study in ethanolic leaf and flower extracts of N. Arbor-tristis revealed the higher anti-oxidant property of leaf than flower. The reducing power increased with increasing concentration of both leaf and flower extract. Studies of similar kind 3 also reported antioxidant activity from N. Arbor-tritis linn. The reducing capacity of a compound may serve as a significant indicator of its potential antioxidant activity 4. This strong reducing power of the plant can also be attributed to the presence of various anti-oxidants such as phenolics and flavonoid contents. The anti-oxidant property of N. Arbor-tristis linn. Is also also due to the presence of carotenoids and lycopene which are potent anti-oxidants. Inflammation is a common phenomenon and it is a reaction of living tissues towards injury. Steroidal anti-inflammatory agents will lyse and possibly induce the redistribution of lymphocytes, which cause rapid and transient decrease in peripheral blood lymphocyte counts to affect longer term response. Ethanolic leaf and flower extracts of N. arbor-tristis Linn. showed a potential in-vitro anti–inflammatory activity on HRBC in agreement with the previous studies. The anti-inflammatory activity may be due to some chemical components present in the leaf and flower such as flavonoids, tannins, coumarins, phenols etc. That are biologically active whose properties might have been found in the ethanolic extract. It has been reported that flavanoids exerts stabilizing effect on lysosomal membrane both in-vitro and in-vivo while tannins possess ability to bind to cations there by stabilizing erythrocyte membranes and other macromolecules 5. N. arbor-tristis leaf and flower contain a number of important phytochemical components which could contribute to medicinally important bioactivities of this plant, including antibacterial activity. The plant materials also show good antioxidant property due to presence of flavonoids, lycopene carotenoids etc. It also possesses anti-inflammatory activity. Time has come to make good use of centuries old knowledge of Nyctanthes arbor tristis Linn. through modern approaches of drug development which has given encouragement among scientists in exploring more information about this medicinal plant. REFERENCE 1] Hanato T, Kagawa H, Yasuhara T, Okuda T; Two new flavonoids and other constituents in lieo rice rout ;Their relative astringency and radical scavenging effect. Chemical and Pharmaceutical Bulletin, 1988; 36:-] Oyedapo OJ, Ekundayo O, Koenig, WA. Leaf volatile oil constituents of Lantana Camara L. from Nigeria. FlavFragr J 2004; 18;-] Meir S, Kanner J, Akiri B, Hadas SP; Determination and involvement of aqueous reducing compounds in oxidative defense systems of various senescing leaves. Journal of Agricultural Food Chemistry, 1995; 43(7):-] Brand-Williams W, Cuvelier M, Berset C; Use of a free radical method to evaluate antioxidant activity. LebensmittelWissenschaft and Technology, 1995; 28(1): 25–30. 5] Havsten, BH. 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