Effect Of H₂S On The Expression Of TGF?1 In Hepatic Stellate Cells

Objective:Investigating the regulating effects of the hydrogen sulfide on the expression of transforming growth factor ?1 protein and transforming growth factor ?1 mRNA of cultured hepatic stellate cells(HSCs)in vitro to provide further theoretical basis for the mechanism of hydrogen sulfide delaying hepatic fibrosis.Methods:1.Culture of HSC-T6 cells:simulate the condition of hepatic fibrosis,hepatic stellate cells were cultured in Petri dishes.HSC-T6 cells were cultured in DMEM/L containing 10%FBS.The cells were adherent after 8 to 10 hours,the medium was aspirted after 36 hours,the cells were subcultured when the proliferation of cells was up to 90% of the growth area in 48 hours and the cells were not used in the experiment until characteristics of the cells was stable.2.The cells were divided into five groups,the experimental grouping according to the dosing concerntration gradient.The NaHS concerntration were 50?mol/L,75?mol/L,100?mol/L and 200?mol/L.The experiment was terminated after aforementioned drugs administration of 48 hours.Using western blot and RT-PCR to detect the expression level of transforming growth factor ?1 protein and transforming growth factor ?1 m RNA.Results:Compared with the control group,different concerntration of the NaHS had significcant effect on the expression of TGF ?1 protein and mRNA in the HSC after 48 hours.Compared with other groups,the expression level of TGF ?1 protein and mRNA decreased significantly in Na HS 50?mol/L group and NaHS75?mol/L group.However,compared with control group,the expression of TGF ?1 protein and mRNA were increased in the Na HS 100?mol/L and 200?mol/L groups.All the differences showed statistically significant differences(P<0.05).Conclusions:1.Exogenous hydrogen sulfide could influence the expression of TGF?1 in HSC-T6 cells.2.Exogenous hydrogen sulfide could regulate the expression of TGF ? 1 in hepatic stellate cells bidirectionally,showing a “bell shaped dose effect”,that means low levels of hydrogen sulfide could inhibit its expression and promote its expression at high concentrations.Hydrogen sulfide delaying hepatic fibrosis may be related to its inhibition of the expression of TGF ?1.