Effects Of Interferon α-1b By Regulati On Of Protein Kinase Cε And Cα In Inhibiting Fibrosis Of Hepatic Stellate Cells And Its Mechanismon

Objective:To investigate the effect of interferon α-1b(IFNα-1b)by regulation of protein kinase Cε(PKCε)and protein kinase Ca(PKCα)in inhibiting fibrosis of hepatic stellate cells and its mechanismon.Methods:Hepatic stellate cell(HSC)-T6 cells were treated with different levels of IFNα-1b and the proliferation of HSC-T6 cells was detected by MTT assay.The expression of PKCε and PKCα was detected by immunofluorescence staining.PKCε,PKCα,β-catenin and Survivin mRNA levels was detected by RT-PCR.PKCε,PKCα,β-catenin and Survivin protein levels was detected by Western blot.Conduct variance analysis using the OneWay ANOVA approach.Results:The inhibition rates of 100,200,400,800 and 1000 U L alpha interferon-1B group were respectively(15.85±1.05)%,(36.59±1.03)%,(45.12±1.05)%,(50.00±:1.01)%and(62.20±1.02)%,with statistically significant inter-group differences(F=27.478,P<0.01);The 48h inhibition rates were(20.87±1.09)%,(43.96±1.08)%,(53.85±1.08)%,(64.84±1.06)%and(74.72±1.07)%respectively,with statistically significant inter-group differences(F=25.321,P<0.01).IC50 was 343.47 Umm L.The levels of hydroxyproline in group 100,200 and 400 U/mL alpha interferon-1b were respectively(7.48±0.28),(6.26±0.17),(3.86±0.20)and lower than that in control group(8.47±0.32).The fluorescence intensity of PKC alpha was also lower than that of the control group.The difference was statistically significant(T values were 9.232,15.921,22.222,all P<0.01).With the increase of the alpha interferon-1b level,the levels of HSC-T6 PKC,PKC alpha,beta-catenin and survivin were significantly lower than those of the control group.With the increase of alpha interferon-1b level,the levels of HSC-T6 PKC,PKC alpha,beta-catenin and Survivin were significantly lower than those of the control group.Conclusion:IFNα-1b can down-regulate the expression of collagen in hepatic stellate cells in a dose-dependent manner,decrease the expression of PKCε,PKCα,β-catenin and Survivin,and inhibit the proliferation of HSC-T6 hepatic stellate cells.Effects of angiotensin Ⅱ on protein kinase Cε and Cα expression in hepatic stellate cellsObjective:To investigate the effect of angiotensin Ⅱ on protein kinase Cε and Cαexpression in hepatic stellate cells.Methods:HSC-T6 cells were treated with different concentrations of angiotensin Ⅱ and the proliferation of HSC-T6 cells was detected by MTT assay.The expression of PKCε and PKCα was detected by immunofluorescence staining.PKCε and PKCα mRNA levels was detected by real time PCR.Results:Angiotensin Ⅱ concentrated the proliferation of HSC-T6 cells(F=25.321,P<0.001)and showed a dose-dependent effect;the level of hydroxyproline(F=13.283,P<0.001).With the increase of Ang Ⅱ concentration,PKCε significantly increased and translocated in the cell membrane;PKCa increased significantly,especially in transplanted membrane and cytoplasm(F=21.387,19.431,P<0.01),and showed obvious dose effect;meanwhile,Increased the expression of PKCε and PKCα,and induced cell proliferation by up-regulating PKCε and PKCa mRNA levels(F=21.387,19.431,P<0.05).Conclusion:Angiotensin Ⅱ can up-regulate the expression of collagen in hepatic stellate cells in a dose-dependent manner,increase the expression of protein kinase Cs and Ca,and promote the proliferation of hepatic stellate cells.