乙型肝炎病毒表面抗原主蛋白与烯酰辅酶A水合酶相互作用对肝细胞胆固醇代谢的影响

Impact of interactions between small hepatitis B virus surface antigen and enoyl co-enzyme A hydratase on cholesterol metabolism in hepatocytes

目的探究乙型肝炎病毒表面抗原主蛋白(SHBs)与烯酰辅酶A水合酶(ECHS1)相互作用对肝细胞胆固醇代谢的影响。方法通过在人肝癌细胞株HepG2中瞬时转染SHBs,建立SHBs瞬时转染细胞模型,分别设置空白对照组、空质粒转染组、单纯SHBs转染组、SHBs+阴性序列对照组、ECHS1过表达组以及ECHS1干扰组。在转染后48 h检测ECHS1及载脂蛋白A1(ApoA1)基因的表达和各转染组转氨酶水平及细胞内总胆固醇(TC)含量。结果与空白组相比,单纯SHBs转染组在核酸水平ECHS1、ApoA1的表达均下降(P均<0.001),蛋白水平表达趋势同核酸一致(P<0.001、P=0.0025),细胞内TC水平升高(P<0.001),转氨酶水平升高(PALT<0.001、PAST<0.001)。较单纯SHBs转染组,ECHS1过表达组可在核酸和蛋白水平下调ApoA1(P=0.0072、P<0.001),且细胞内TC水平下降(P<0.001),转氨酶水平降低(PALT<0.001、PAST=0.0049)。与单纯SHBs转染组相比较,ECHS1干扰组Apo A1核酸和蛋白水平亦降低(P=0.0096、P<0.001),TC水平降低(P<0.001),ALT水平无显著变化(P=0.28),AST水平升高(P=0.0069)。结论 SHBs能够通过下调ApoA1表达来提高肝细胞TC水平,同时ECHS1通过相关信号通路及分子机制抑制TC的生成。

Objective To investigate the interactions between small hepatitis B virus surface protein(SHBs)and enoyl CoA hydratase short chain 1(ECHS1)on cholesterol metabolism in hepatocytes.Methods Human hepatocellular carcinoma HepG2 cells were transfected with SHBs transiently,and then assigned into the blank control group,the blank plasmid transfection group,the SHBs transfection group,the SHBs+negative sequence control group,the ECHS1 overexpression group and the ECHS1 interference group.The expression of ECHS1 and apolipoprotein A1(ApoA1)were detected,and the levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST)and intracellular total cholesterol(TC)were measured 48 hours post-transfection.Results Significantly lower ECHS1 and ApoA1 expression were detected in the SHBs transfection group than those of the blank control group at both translational(both P<0.001)and transcriptional levels(P<0.001,P=0.0025),and higher intracellular TC(P<0.001)and serum ALT(P<0.001)and AST levels(P<0.001)were measured in SHBs transfection group than those of the blank control group.In addition,lower ApoA1 protein(P=0.0072)and mRNA expression(P<0.001),lower intracellular TC levels(P<0.001),lower serum ALT(P<0.001)and AST concentrations levels(P=0.0049)were found in the ECHS1 overexpression group than those in the SHBs transfection group;meanwhile,compared with SHBs transfection group,there were significantly lower ApoA1 nucleic acid and protein levels(P=0.0096,P<0.001),decreased TC level(P<0.001),no significant change of ALT level(P=0.28),and increased AST level(P=0.0069)in ECHS1 interference group.Conclusions SHBs increases TC levels through downregulating ApoA1 expression in hepatocytes,and ECHS1 may suppress TC production through some signal pathways and molecular mechanism.