摘要
目的 观察乙型肝炎病毒X蛋白（HBx）对1α，25-（OH）2D2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＋1d，25（OH）2D3组、共转染pEGFP-N1＋pcDNA3-X-＋1α，25（OH）2D3组。同时以0．35μmol／L1d，25（OH）2Ds诱导HepG2细胞CYP3A4表达72h，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＋pcDNA3X＋1d，25-（OH）2D3组CYP3A4mRNA表达量是空白对照组的1．52倍，而转染pEGFP-N1＋1α，25（OH）2D。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）2D。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。
Objective To investigate the effect of hepatitis B virus X protein （HBx） on the induction of eytochrome P450 （CYP） 3A4 by 1α,25 （OH）2 D3 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 transfeetion）, plasmid pEGFP-N1 transfection group, plasmicl pEGFP-N 1 transfeetion plus 1α, 25- （OH）2D1α group, plasmid pcDNA3-X and pEGFP- N1 co transfection plus 1α,25 （OH）2D2 group. The expression of CYP3A4 in HepG2 cell was induced by 0.35 /,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 transfecti0n plus 1α,25-（OH）2D2 group was 3.97 folds （F=4.72, P〈0. 05）. Similarly, intracellular CYP3A4 protein expression in plasmid pcDNA3-X and pEGFP-N1 co-transfeetion 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）2D2 group increased to 5.9 folds （F= 4.68, P〈0.05）. Conclusion HBx interferes with the induction of CYP3A4 by 1α, 25 （OH）2D2, in HepG2 cell line, which suggests that HBx has suppressive effect on the expression of CYP3A4.
出处
《中华传染病杂志》
CAS
CSCD
北大核心
2009年第8期449-453,共5页
Chinese Journal of Infectious Diseases
基金
国家自然科学基金资助项目（30571660）
