Isolation, identification and characterization of apigenin fromJusticia gendarussaand its anti-inflammatory activity

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Inflammatory responses during chronic diseases such as atherosclerosis, cancer etc., are harmful to host organisms. Generally NSAIDs are used to treat against these severe conditions but due to its adverse effects studies are going on with medicinal plants, since they are rich in bioactive compounds. Justicia gendarussa is one such plant which has been used as a remedial measure for treating inflammatory diseases since ancient time. Thus the present study involved in the isolation, characterization and identification of apigenin (flavonoid) from this plant and to elucidate its molecular mechanism against inflammation via TLR-NF-κB signaling pathway using ox-LDL induced hPBMCs in in vitro model. Methanolic extract was used for the isolation process and results showed that the F6 fraction collected from ethyl acetate through column chromatography showed 89% paw edema inhibition at a dose of 10mg/kg in carrageenan induced rats. Purification of F6 by TLC with toluene: chloroform: acetone (8:5:7) and further characterization by 1HNMR indicated the presence of bioactive compound, apigenin. In vitro studies revealed that pretreatment of ox-LDL induced hPBMCs with apigenin (25μM) significantly (P<0.05) reduced the levels of TLR4, MyD88, TRIF, TRAF6, NF-κB, COX-2, PGE2, IL-1β and TNF-α responsible for generating inflammation and elevated the level of anti-inflammatory cytokine, IL-10. These results indicate the therapeutic efficacy of bioflavonoid apigenin which was isolated from Justicia gendarussa against ox-LDL induced inflammation. Therefore apigenin can be treated as a suitable therapeutic agent against inflammatory diseases.HIGHLIGHTSF6 Ethyl acetate fraction of Justicia gendarussa showed 89% paw edema inhibition in carrageenan induced rats.Apigenin downregulated TLR 4 signaling cascade by inhibiting the activation of adapter proteins.Inhibition of ox-LDL induced COX-2 expression by apigenin indicates its anti-inflammatory effect.Apigenin imparts atheroprotective effect by reducing the levels of pro-inflammatory genes.

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