Effects of exogenous transforming growth factor-β3 on the expression of endogenous transforming growth factor-β3 and proliferation of hepatic stellate cells / 中华消化杂志

ABSTRACT

Objective To investigate the effects of exogenous transforming growth factor-β3 (TGF-β3) on the expression of endogenous TGF-β3 and proliferation of rat hepatic stellate cells (HSC).MethodsRat HSC cells were seeded in 24-well plates and were divided into 2 groups.One group of cells were exposed to TGF-β3 of different concentrations (0.08,0.4,2,10 and 50 ng/ml) for 2 hours and then the cell culture supernatant was collected; the other group of cells were exposed to 10 ng/ml TGF-β3 and the cell culture supernatant was collected at different time point (0.25,0.5,1,2,4,8 and 13 h).The content TGF-β3 was determined by ELISA method.Some other HSC cells were seeded in 96-well plates and divided into 2 groups.One group of cells were cultured in the presence of exogenous TGF-β3 of different concentrations (0.001,0.005,0.02,0.08,0.32,1.25,5,20,100,500 ng/ml) for 24 hours and then the cell proliferation was detected; the other group of cells were treated with 5 ng/ml TGF-β3 for 24 and 48 hours.The cell proliferation was measured by MTT method.The HSC cell morphology of control group and TGF-β3 treated group was observed under inverted microscope. Results The endogenous TGF-β3 expression of HSC cells obviously increased after exogenous TGF β3 treated at 2 ng/ml and reached the peak at 3 hour [(0.845±0.028) ng/ml vs (0.026±0.021) ng/ml,F=210.168,P=0.00].Low concentrations of exogenous TGF-β3 did not affect the proliferation of HSC cells.Above 0.32 ng/ml,exogenous TGF-β3 could promote HSC proliferation.There was no dose-dependent relationship between HSC cell proliferation and the concentration of exogenous TGF-β3 (F=0.68,P=0.57).Under microscope,lhere was no significant difference in HSC cell morphology between control group and TGF-β3 treated group.Conclusions Exogenous TGF-β3 can promote the expression of endogenous TGF-β3 in HSC cells,and can promote HSC cell proliferation.There is no obvious effect of exogenous TGF-β3 on HSC morphology.