摘要目的 观察乙型肝炎病毒X蛋白(HBx)对1α,25-(OH)2D3诱导的HepG2细胞色素P450(CYP)3A4的影响.方法 将重组HBx真核表达质粒pcDNA3-X与pEGFP-N1质粒(转染对照)瞬时共转染HepG2细胞,分为空白细胞对照组(对照组)、转染pEGFP-N1组、转染pEGFP-N1+1α,25-(OH)2D3组、共转染pEGFP-N1+pcDNA3-X+1α,25-(OH)2D3组.同时以0.35 μmol/L1α,25-(OH)2D3诱导HepG2细胞CYP3A4表达72 h,分别以RT-PCR法和Western印迹法,在mRNA水平和蛋白水平检测CYP3A4的表达.组间比较行F检验.结果共转染pEGFP-N1+pcDNA3-X+1α>,25-(OH)2D3组CYP3A4 mRNA表达量是空白对照组的1.52倍,而转染pEGFP-N1+1α,25-(OH)2D3组为3.97倍(F=4.72,P<0.05);共转染pEGFP-N1+pcDNA3-X+1α,,25-(OH)2D3组CYP3A4蛋白诱导倍数是2.1,而转染pEGFP-N1+1α,25-(OH)2D3组是5.9(F=4.68,P<0.05).结论 HBx干扰1α,25-(OH)2D3对HepG2细胞CYP3A4的诱导,提示HBx对CYP3A4的表达调控有抑制作用.

abstractsObjective To investigate the effect of hepatitis B virus X protein (HBx) on the induction of eytochrome P450 (CYP) 3A4 by 1α, 25-(OH)2D3 in HepG2 cells in vitro. Methods HepG2 cells were transiently transfected with plasmid pEGFP-N1 (control) or co-transfected with recombinant HBx eukaryotic expression plasmid pcDNA3-X and pEGFP-N1. All HepG2 cells were divided into four groups: control group (without transfection), plasmid pEGFP-N1 transfection group, plasmid pEGFP-N1 transfection plus 1α ,25-(OH)2D3 group, plasmid pcDNA3-X and pEGFP-N1 co-transfection plus 1α ,25-(OH)2D3 group. The expression of CYP3A4 in HepG2 cell was induced by 0.35 μ mol/L 1α ,25-(OH)2D3 for 72 h, and mRNA levels and protein levels of CYP3A4 in the cells were detected by reverse transcriptase-polymerase chain reaction (RT-PCR) assay and Western-blot assay, respectively. The comparison between groups was done by F test. Results CYP3A4 mRNA level in plasmid pcDNA3-X and pEGFP-N1 co-transfection plus 1α ,25-(OH)2D3 group was 1.52 folds of control group, while that in plasmid pEGFP-N1 transfection plus 1α, 25-(OH)2D3 group was 3.97 folds (F= 4.72, P<0. 05). Similarly, intracellular CYP3A4 protein expression in plasmid pcDNA3-X and pEGFP-N1 co-transfection plus 1α , 25-(OH)2D3 group increased to 2.1 folds of control group, while that in plasmid pEGFP-N1 transfection plus 1α,25-(OH)2D3 group increased to 5.9 folds (F=4.68, P<0.05). Conclusion HBx interferes with the induction of CYP3A4 by 1α , 25-(OH)2D3 in HepG2 cell line, which suggests that HBx has suppressive effect on the expression of CYP3A4.