Protective effect of ethyl acetate fraction of Drynaria quercifolia against CCl4 induced rat liver fibrosis via Nrf2/ARE and NFκB signalling pathway

Ethnopharmacological relevance: Drynaria quercifolia rhizome is traditionally used as hepatoprotective drug especially in chronic jaundice. Aim of the study: The present study was undertaken to scientifically evaluate the efficacy of D. quercifolia rhizome against liver fibrosis. Materials and methods: D. quercifolia rhizome crude extract (DQ) and its fractions of hexane (HDQ), ethyl acetate (EDQ), butanol (BDQ) were evaluated in vitro using primary hepatocytes and RAW 264.7 cells. In vivo anti-liver fibrotic activity of EDQ was assessed using CCl4 induced liver fibrosis in Wistar rats and serum biochemical parameters (AST, ALT, ALP, SB, cholesterol), MDA, PT, INR, GSH, SOD, CAT, liver glycogen, serum albumin levels were monitored. qRT-PCR analysis of TNF-alpha, COX-2, iNOS were performed. ELISA method was used to estimate TNF-alpha, COX-1 & 2. Histopathological studies like H & E, Masson's trichrome, immunohistochemistry staining for alpha-SMA, TIMP-1, Nrf2 were conducted. LC-Q-TOF-MS analysis of EDQ was conducted. Results: In vitro activity guided fractionation of D. quercifolia revealed EDQ as active fraction when compared to other extracts. EDQ treatment significantly inhibited the expression of alpha-SMA, TIMP-1, COX-2, TNF-alpha, iNOS and increased the levels of Nrf2 in rat liver fibrosis. LC-Q-TOF-MS analysis of EDQ confirmed the presence of naringin and naringenin. Conclusion: The anti-liver fibrotic activity of EDQ is via inhibition of NF kappa B signalling pathway, antioxidant response through Nrf2 activation and further inhibition of HSC activation.