In vivo and in-vitro anti-inflammatory study

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  • 1. Article Reference Number: OPP 029 In-vitro and In-vivo anti-inflammatory potential of Tabernaemontana divaricata leaves Sumithra Mohan*, J. Anbu, Arijit Chakraborty, Ariadasy Canagasaby, Odelia Sawian School of Pharmaceutical Sciences, Vels University, VISTAS Pallavaram, Chennai -117. Presenting author Arijit Chakraborty M.Pharm (Pharmacology) Vels University
  • 2. Inflammation Definition • Inflammation is a non specific, localized immune reaction of the organism, which tries to localized the pathogenic agents. Many consider the syndrome a self- defense mechanism. • It consist in vascular, metabolic, cellular changes, triggered by the entering of pathogenic agents in healthy tissues of the body.
  • 3. Mechanism action of Anti-inflammatory drugs. Arachidonic acid COX 1 Constitutive GI Prostagland mucosa ins GI mucosal protection COX 2 Inhibitors Platelet Thrombox ane Hemostasis Prostaglan dins Mediate pain, inflammation and fever
  • 4. Plant selection Ta be rna e m o nta na d iva ric a ta (Family - Apocynaceae) is a beautiful evergreen shrub, about 54cm high, with large shiny leaves, crepe jasmine flowers, may appear sporadically all year. It , is a rich source of alkaloids with various pharmacological Kingdom Plantae properties. Order Gentianales Family Apocynaceae Subfamily Rauvolfioideae Genus Tabernaemontana Species T. d iva ric a ta
  • 5. Working procedure
  • 6. In-vitro model Ref: Kar B; Suresh Kumar RB; Karmakar I; Dolai N, Bala A; Mazumder UK, Haldar PK; Antioxidant and in vitro antiinflammatory activities of Mimusops elengi leaves; Asian Pacific Journal of Tropical Biomedicine; Vol. 2012; pp S976S980
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  • 9. The percentage of HRBC membrane stabilization or protection was calculated using the formula: O.D of Test Solution – O.D of Product Control Percentage Inhibition of Haemolysis = 100 – ------------------------------------------------------------ ×100 Control O.D of Test
  • 10. Observation table for In-vitro study Table 1: % Prevention of Lysis Concentration (µg/ml) Extract Diclofenac sodium 100 50 25 15.5 48.14 47.58 47.07 47.36 42.1 36.84 31.54
  • 11. Effect of concentration on % prevention of lysis of extract treated group Effect of concentration on % inhibition of lysis of diclofenac treated group
  • 12. In-vivo model Ref: Winter CA; Risley EA; Nuss GW; Carrageenin induced oedema in hind paw of rat as an assay for anti-inflammatory drugs; Proc Soc Exp Biol Med 1962; 111; EETD*: Ethanolic extract of Ta be rna e m o nta na d iva ric a ta Continue… …..
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  • 15. Observation table for In-vivo study Table 2: TREATMENT DOSE (mg/kg) MEAN INCREASE IN PAW VOLUME (ML) PERCENTAGE INHIBITION Control 5 ml/kg 0.778± 0.056 - Diclofenac sodium 5 mg/kg 0.153± 0.034 58.13 EETD* 400 mg/kg 0.277± 0.023 42.19 EETD* 0.325± Ta be 36.02 EETD*: 200 mg/kg Extract of0.081 rna e m o nta na Ethanolic d iva ric a ta .
  • 16. Effects of dose on mean increase in paw volume Effect of dose on percentage inhibition
  • 17. Result for In-vitro study (Table 1) : • The extract at concentration range of 6.3 -100 mg/ml protect the human erythrocyte membrane against lysis induced by hypotonic solution. • At concentration of 100µg/ml, the extract produced 47.36 % inhibition of RBC haemolysis as compared with 48.14% produced by diclofenac sodium. Result for In-vivo study (Table 2) : • The extract at concentration 200 mg/kg and 400 mg/kg protect the human erythrocyte membrane against carrageen induced. • At concentration 200 mg/kg and 400 mg/kg, the extract produce 36.02 % and 42.63 % inhibition of inflammation as compared with 58.13 % produced by diclofenac sodium.
  • 18. Conclusion In conclusion the present study demonstrated that Ta be rna e m o nta na d iv a ric a ta leaves extract has anti-inflammatory activity at a dose of 100 µg/ml in in-vitro study and 200 mg/kg and 400 mg/kg in in-vivo study.
  • 19. Acknowledgement The authors are thankful to the authority of the School Of Pharmaceutical Science, Vels University, Pallavaram, Chennai 600117, India, for providing necessary facilities for the present study.