Effects Of Sp600125 On Acetaldehyde-induced Proliferation And Apoptosis Of Hepatic Stellate Cells And Protein Expression Of Bcl-2, C-myc And Caspase-3 In Rats

Background and aimsIt's showed that the activation of hepatic stellate cell(HSC) was the key element in the progress of hepatic fibrosis.The proliferation of HSC induced by acetaldehyde was the key factor in alcoholic hepatic fibrosis.In recent years,many studies indicated that mitogen-activated protein kinase(MAPK) including ERK,JNK and P38 was one of the main signal pathway in the activation and proliferation of HSC.In the recent study,we found that the phosphorylation of JNK decreased with sp600125 concentration increasing in acetaldehyde-induced HSC by western blot.This experiment was to study the effects of sp600125 on the proliferation and apoptosis,the protein expression of bcl-2,c-myc and caspase-3 in acetaldehyde-induced HSC-T6,aimed to explore the mechanism of alcoholic hepatic fibrosis.MethodsThe acetaldehyde-induced HSC-T6 was treated with different doses of sp600125.The proliferation of HSC-T6 was evaluated by MTT colorimetric assay,and the morphological changes of HSC-T6 were observed by Hoechst 33258 staining.The cell cycle proportion and apoptotic rate of HSC-T6 were analyzed by flow cytometry(FCM),and the protein expression of bcl-2,c-myc and caspase-3 were examined by SABC method.ResultsThe proliferation of HSC-T6 was inhibited by different doses of sp600125(F = 102.526,P＜0.01),and with dose-dependent relationship(r = -0.928,P＜0.01).With sp600125 concentration increased,the cell proportion of G₀/G₁ stage increased gradually(F=6.335,P＜0.01),S stage decreased by degrees(F=39.202,P＜0.01),so did the cell proliferation index(F=7.043,P＜0.01),the apoptotic rate of HSC-T6 significantly increased(F = 54.834,P＜0.01) as well as;bcl-2 and c-myc protein expression positive rate decreased(F value were 227.084 and 418.931,P＜0.01),meanwhile caspase-3 increased (F= 38.259,P＜0.01).Conclusionsp600125 could inhibit HSC-T6 proliferation and accelerate apoptosis,which was possibly related to block HSC-T6 from G₀/G₁ to S and down-regulation the protein expression of bcl-2 and c-myc,as well as up- regulation the protein caspase-3.