| Background Although liver fibrosis is a disease which badly threatens people's health, no effective therapy exists at present. Excess deposition of extra-cellular matrix proteins (ECM) is the main performance in liver fibrosis. Fibrosis is the early stage of cirrohosis, accompanied with activation and proliferation of HSCs. It has been proved that TGF-β1 is a strong pro-fibrogenic cytokine. TGF-β1 and its downstream signal CTGF plays an essential role in the process of hepatic fibrosis.Clinical epidemiological data showed that for liver cirrhosis and its complications, gender differences existed. Liver fibrosis in the cellular and animal model had also been confirmed the fibrosuppressive of eatrogen .Estrogen fulfills its important physiological function through the interaction with ER, but it still has side effects. Recent studies showed that estrogen metabolites could protect the heart and kindy from oxidation damage and inhibit the proliferation of endotheial cells, concluding that the fibrosuppresive role of estrogen may be through the metabolites. The scholars at home and abroad had confirmed estrogen metabolites can not bind to ER, so that they couldn't produce feminine effect. We assumed that it could be applied to either male or female.AIM The present study was undertaken to evaluate the proliferation and the expression of transforming growth factor-pi (TGF-β1) and connective tissue growth factor (CTGF) of hepatic stellate cells incubated with estradiol metabolites: 2- hydroxyestra-diol(2OHE), 4-hydroxyestradiol(4OHE), 2-methoxyestradiol(2MeOE), investigating the mechanism of hepatic fibrosuppresive of estradiol and its metabolites, to provide a stronger theoretical basis of application of estradiol and its metabolites on clinical treatment of liver fibrosisMethods The hepatic stellate cells (LX-2)were randomly divided into 13 groups, and were added with different concecentraton of estradiol and its metabolites(2OHE,4OHE,2MeOE),and at the same time,a positive control was set up .After that ,the cells contributed to be incubated for 48 hours, and at the end point ,the patemeters below were detected: 1) the proliferation of LX-2 by MTT asssy. 2)The expression of TGF-β1 and CTGF mRNA by RT-PCR. 3)The expression of TGF-β1 and CTGF protein by cellular immunochmeistry .Results 1) Among the conceration of 10-9M～10-7M of estradiol and its metabolites , the inhibition of estradiol metabolites on LX-2 proliferation had a does-dependant manner(p=0.000). Compared with the same concentration, there was significant difference between estradiol metabolites and estradiol (P<0.05). 2MeOE had the strongest biological activation, and there was no significant difference between 2OHE and 4OHE. 2) Among the conceration of 10-9M～10-7M of estradiol and its metabolites , the expression of TGF-β1 and CTGF mRNA decreased, which also had a does-dependant manner (p=0.000); there was significant difference between estradiol metabolites and estradiol(P<0.05). 2MeOE had the strongest biological activation (P<0.05), and there was no significant difference between 2OHE and 4OHE (P>0.05). 3) At the concentration of 10-7M of estradiol and its metabolites , the expression of TGF-β1 and CTGF protein was inhibited and the inhibition activity of 2MeOE is the strongest(P>0.05), but there was no difference between 2OHE and 4OHE.(P>0.05)Conclusion 1)Estradiol metabolites can inhibit the proliferation of LX-2. 2) Estradiol metabolites can inhibit the expression of HSCs TGF-β1 mRNA and its downstream signal-CTGF mRNA .3) Compared with ordinary estradiol, the estradiol metabolites have the stronger inhibition on the proliferation and the expression of TGF-β1 and CTGF of LX-2. 2MeOE has the strongest biological effect in estradiol metabolites.
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