Neferine inhibits cultured hepatic stellate cell activation and facilitates apoptosis A possible molecular mechanism

Neferine is a major alkaloid component of Lian Zi Xin embryos of the seeds of Nelumbo nucifera Gaertner Nymphaeaceae Previous studies have shown that neferine ha an inhibitory effect on pulmonary fibrosis through its anti-inflammatory and anti oxidative activities and inhibition of cytokines and NF-kappa B However it is unknown whether neferine also has an inhibitory effect on liver fibrosis through inhibition of TGF-beta 1 and collagen land facilitation of apoptosis of hepatic stellate cells This study examined the effects of neferine on cultured hepatic stellate (HSC-T6) cells and explored its possible action mechanisms by means of MTT a say enzyme-linked immunosorbent assay flow-cytometric annexin V-PI assay and Hoechst 33258 staining as well as real-time PCR and western blotting The results showed that neferine administration (2 4 6 8 and 10 mu mol/l) significantly decreased the TGF-beta 1 and collagen I produced in HSC T6 cells and Increased the HSC 16 cell apoptosis in a dose-dependent manner Neferine treatment for 48 h at concentrations of 6 and 10 mu mol/l significantly increased Sax and caspase 3 mRNAs and proteins and reduced Bcl2 and alpha smooth muscle actin (alpha-SMA) mRNAs and proteins Our data indicate that neferine efficiently inhibits cultured HSC-T6 cell activation and Induces apoptosis by increasing Bax and caspase 3 expression via the mitochondrial pathway (C) 2010 Elsevier B V All rights reserved