Inhibition effect of phytoestrogen calycosin on TGF-β1-induced hepatic stellate cell activation, proliferation, and migration via estrogen receptor β

The present study was designed to investigate the effects of calycosin on hepatic stellate cell (HSC) function and to explore whether the drug exerts its effect through the estrogen receptor. HSC proliferation and migration were measured by MTT assay and transwell chamber assay, respectively. The mRNA and protein expression of alpha-SMA, COL-I, and ER beta were detected by real-time PCR and Western blotting. The co-localization and expression of alpha-SMA and ER beta protein were detected by immunofluorescence. All the studies were investigated in the absence or presence of ICI 182,780. The results showed that calycosin inhibited the proliferation of activated HSCs and remarkably inhibited HSC migration. Calycosin significantly reduced the expression of alpha-SMA and COL-I in activated HSCs. However, with co-treatment with ICI 182,780, the inhibitory effect of calycosin against the above effects was strongly negated. Importantly, calycosin significantly downregulated the expression of ER beta protein, while co-treatment with ICI 182,780 partially reversed the ER beta downregulation. In addition, alpha-SMA decreased with the decrease of ER beta expression and the subtype of ER beta on HSC is ER beta 5. In conclusion, calycosin inhibits proliferation, activation, and migration of TGF-beta(1)-induced HSCs. The effect may be related to binding and downregulation of ER beta 5.