Vijayakumar and Hindumathy
ISSN: 2321-5674(Print) ISSN: 2320 – 3471(Online) Indian Journal of Research in Pharmacy and Biotechnology
INVITRO ANTI-INFLAMMATORY ACTIVITY OF STRYCHNOS POTATORUM LINN SEED BY HRBC MEMBRANE STABILIZATION V.Vijayakumar*1, Dr C.K.Hindumathy 2 1. Department of Pharmaceutical Chemistry, Vinayaka Missions College of Pharmacy. 2. Department of Biotechnology, V.M.K.V. Engineering College. * Corresponding author: E-mail:
[email protected] ABSTRACT In the present study, strychnos potatorum linn belonging to the family Loganiaceae (Strychnaceae) was studied. It is a folkloric medicinal plant used to treat antibacterial, antifungal, anti diabetic, antioxidant and anti-inflammatory. Since many flavonoids have remarkable anti-inflammatory activity the present work aims at evaluating the anti inflammatory activity of seeds strychnos potatorum linn by HRBC membrane stabilization method. The prevention of hypo tonicity induced HRBC membrane lysis was taken a measure of anti inflammatory activity. The anti-inflammatory activity of hydroalcholic extract was comparable to that of the standard drug Hydrocortisone. The percentage protection for the hydroalcholic extract and hydrocortisone were 100 at µg/ml. the hydroalcholic extract of strychnos potatorum linn seed has significant anti-inflammatory activity (54.95±0.74). Key words: Anti-inflammatory, strychnos potatorum linn, HRBC membrane stabilization, Hydroalcholic extract. INTRODUCTION Inflammation is a normal protective response to tissue injury that is caused by physical trauma, noxious chemicals or microbiological agents. Inflammation is the result of concerted participation of more number of proliferative factors (like Vasoactive, chemotatic) at different stages and there are many targets for anti inflammatory activity. (Tripathi, K.D, 2004). The irrespective tissue injury is a type of inflammatory response suppressed by glucocorticoids and this is the basic clinical uses and also it interferes with several steps in the inflammatory response. Numbers of corticoids are only palliative; do not act on the inflammation instead of they favor spread of infections capacity of defensive cells to kill microorganisms is impaired at the same time interfere with healing and scar formation. The alternate use of corticoids is hazardous other than the corticosteroids the NASIDs are also used to treat inflammation .the main mechanism of action of the NASIDs are the inhibition of prostaglandin (PG) synthesis or preferential or selective COX-2 inhibition. Due to the inhibition of prostaglandin (PG) synthesis it may produce toxic effects like bleeding, inhibition of platelet function, gastric mucosal damage, asthma and anaphylactic reactions may cause some individuals. (Tripathi, K.D, 2004).the plant strychnos potatorum linn commonly known as Nirmali and otherwise called clearing nut. The plant is distributed throughout India, Ceylon, Burma, and Deccan. The present work aims at evaluating the anti inflammatory activity of strychnos potatorum linn seed by HRBC membrane stabilization. MATERIALS AND METHODS Plant material: The fresh seeds of strychnospotatorum Linn were collected from shervaroys hills, Salem district, Tamilnadu, India. The collected seeds were identified and authenticated by the Botanist Dr. A. Balasubramanian (consultant central siddha research) Executive Director ABS Botanical Garden, Salem, Tamilnadu. Preparation of extract: The dried seeds are powdered using mixer grinder; 1000gm of the powdered seeds was packed evenly in the soxhelet extractor and subjected to extraction with Hydroalchol (70% ethanol, 30% water).After extraction, the solvent was distilled off and the extracts were concentrated on water bath to a dry residue and kept in a desiccator. Anti inflammatory activity: The HRBC membrane stabilization has been used as a method to study the antiinflammatory activity. (Gandidasan.R, 1991) Blood was collected from healthy volunteers. The collected blood was mixed with equal volume of sterilized Alsever solution (2% dextrose, 0.8% sodium citrate, 0.05% citric acid and 0.42 % sodium chloride in water).the blood was centrifuged at 3000 rpm and packed cells were washed with isosaline (0.85 % , pH 7.4) and a 10% v/v suspension was made with isosaline. The assay mixture contains the drug (at various concentrations as mentioned in the table), 1 ml phosphate buffer (0.15 M, pH 7.4), and 2 ml of hypo saline (0.36%) and 0.5 ml of HRBC suspension. Hydrocortisone sodium was used as the reference drug. Instead of hypo saline 2 ml of distilled water was used in the control. All the assay mixtures were incubated at 370C for 30 minutes and centrifuged. The haemoglobin content in the Volume 1(3)
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ISSN: 2321-5674(Print) ISSN: 2320 – 3471(Online) Vijayakumar and Hindumathy Indian Journal of Research in Pharmacy and Biotechnology supernatant solution was estimated using spectrophotometer at 560 nm. The percentage of hemolysis was calculated by assuming the hemolysis produced in the presence of distilled water as 100%. The percentage of HRBC membrane stabilization or protection was calculated by using the following formula,
RESULTS AND DISCUSSION The lysosomal enzymes released during inflammation produce a various disorders. The extra cellular activity of these enzymes is said to acute or chronic inflammation. The non steroidal drugs act either by inhibiting these Lysosomal enzymes or by stabilizing the membrane. (Rajendran Vadivu, 2008) since HRBC membrane are similar to Lysosomal membrane components the prevention of hypo tonicity induced HRBC membrane lysis is taken as a measure of anti inflammatory activity of drugs. The results were reported in table 1. It was observed from the table 1 and figure 1 that the hydroalcholic extract shows significant anti inflammatory activity at the concentration 100 µg/ml, which is comparable to the standard drug hydrocortisone sodium. The anti inflammatory activity of the extract were concentration dependent, with increasing concentration the activity is also increased. The hydroalcholic extract has significant anti inflammatory activity. Table.1. In-vitro anti inflammatory activity strychnos potatorum linn seed Concentration in Activity ( % protection ) Standard Hydrocortisone ( µg/ml ) sodium HAESP Control 20
43.53±0.62
55.48±0.57
40
46.41±0.87
57.71±0.54
60
49.00±0.50
59.21±0.95
80
52.57±0.68
63.56±0.68
100
54.95±0.74
67.50±0.52
(Values are expressed as SEM of 3 readings) Figure1. In-vitro anti inflammatory activity strychnos potatorum linn seed
Activity % protection
Invitro anti inflammatory effect of strychnos potatorum linn 80 60 40
HAESP
20
STANDARD DRUG
0 20
40
60
80
100
concentration in µg /ml
CONCLUSION The extract exhibited membrane stabilization effect by increasing hypo tonicity induced lysis of erythrocyte membrane. The erythrocyte membrane is analogus to the Lysosomal membrane (Chou, 1977) and its stabilization implies that the extract may as well stabilize Lysosomal membrane. Stabilization of Lysosomal membrane is very important in limiting the inflammatory response by preventing the release of Lysosomal substances of activated neutrophil such as bacterial enzymes and proteases which cause further tissue damages. (Murugasan, 1981) From the investigation it was concluded that the hydroalcholic extract of strychnos potatorum linn has significant membrane stabilization property and it was comparable to the standard drug hydrocortisone. Volume 1(3)
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Vijayakumar and Hindumathy
ISSN: 2321-5674(Print) ISSN: 2320 – 3471(Online) Indian Journal of Research in Pharmacy and Biotechnology REFERENCES
Chou CT, The Anti inflammatory effect of Tripterygium Wilfordihook on adjuvant induced paw edema in rats and inflammatory mediator’s release, Phytother Res, 11, 1997, 152-154 Gandidasan R, Thamaraichelvan A, Baburaj S, Anti inflammatory action of Lannea coromandelica by HRBC membrane stabilization. Fitoterapia, 1991, voll LXII, 81-83. Kirtikar K.R, Basu B.D, Indian Medicinal plants. International book distributors, Dehradum.1975; II edn; Vol III; 1971. Murugasan, Vember, S, Damodharan, C, C. Studies on erythrocyte membrane IV, In-vitro hemolytic activities of Oleander extract .Toxicol Lett, 8, 1981, 33-38. Nirmala Devi K, Periyanayagam K, in-vitro Anti inflammatory activity of Plectrnthus Amboinicus (Lour) spring by HRBC membrane stabilization, International journal of pharmaceutical studies and Research, 1(1), 2010; 2629. Rajendran Vadivu, Lakshmi K.S.In vitro and in-vivo anti inflammatory activity of Leaves of Smplocos cochinchinenisis (Lour) Moore ssp Laurina. Bangladesh JPharmacol, 3, 2008, 121-124. Ram P Rastogi, Mehrotra B N, Compendium of Indian Medicinal plants, CDRI Lucknow and publication and information Directorate, New Delhi, 2, 1970, 79-201. Tripathi KD, Essentials of Medical Pharmacology, Jaypee brothers medical publishers (P) Ltd, New Delhi, 2004, V edn; 167-181, 257-259. Warrier P.S, Indian Medicinal plants, Arya Vaidhya Sala, Kottakkal, Orient Longmann limited. Hyderabad, Voll IV, 315-317.
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