Effect of Loureirin A on Proliferation and Frizzled-4 Expression of Rat Hepatic Stellate Cells in vitro

［Abstract］Objective　To investigate the molecular mechanism of Loureirin A mediated anti-hepatic fibrosis by evaluting its effects on proliferation ，secretion of α-smooth muscle actin（α-SMA）and transforming growth factor-beta1（TGF-β1），and expression of rat hepatic stellate cells in vitro . Methods Primary hepatic stellate cells were isolated and cultured from Sprague-Dawley rats. After activating and inducing primary hepatic stellate cells from qHSC to aHSC，the activated hepatic stellate cells model in vitro was established. Then we observed the morphological changes of static hepatic stellate cells and activated hepatic stellate cells with inverted phase contrast microscope. Cultured hepatic stellate cells were treated with different concentrations of loureirin A and the inhibitory rate of HSCs proliferation was measured by MTT assay. The expression of Frizzled-4 was measured by western blot analysis. The content of α-SMA and TGF-β1 in the cultured HSCs'supernatant were measured by enzyme-linked immunosorbent assay（ELISA） . Results Loureirin A the proliferation of inhibited activated hepatic stellate cells in a time-dose-dependent manner compared with the control group，IC50=0.30 μg/μL. After loureirinA treatment of the HSCs，western blot analysis showed that Frizzled-4 expression level was obviously lower than control group. Loureirin A also inhibited α-SMA and TGFβ1（P＜0.05）secretion in the cultured HSCs'supernatant in different degree by the assay of ELISA. Conclusions The molecular mechanism of Loureirin A and Wnt signaling pathway mediated anti-hepatic fibrosis and anti-angiogenesis may involve down-regulation the expression of Frizzled-4，inhibiting the synthesis and secretion of α-SMA，TGF-β1and the proliferation of HSCs.