检测乙型肝炎病毒对载脂蛋白E表达的影响

Study on the effect of hepatitis B virus on the expression of apolipoprotein E

目的探讨乙型肝炎病毒(HBV)在细胞水平对载脂蛋白E(Apo E)表达的影响。方法采用逆转录聚合酶链式反应(RT-PCR)检测Hep G2.2.15细胞(整合了HBV全基组)及其对照细胞Hep G2中Apo E mRNA表达水平的差异;采用脂质体转染试剂Lipofectin2000将HBV感染性克隆p HBV1.3转染Hep G2细胞,并设立转染空载体p Blue-ks的Hep G2细胞作为空白对照,转染48 h后,采用RT-PCR检测两者Apo E mRNA表达水平的差异,并采用全自动生化分析仪Olympus AU5400检测Hep G2细胞上清中Apo E的含量。结果整合了HBV全基组的Hep G2.2.15细胞中Apo E mRNA的表达水平明显低于对照细胞Hep G2;与转染p Blue-ks的Hep G2细胞相比,转染p HBV1.3的Hep G2细胞中Apo E mRNA的表达显著降低,同时,转染p Blue-ks的Hep G2细胞上清中Apo E的含量为4.45 g/L±1.62 g/L,而转染了p HBV1.3的Hep G2细胞上清中Apo E的含量为3.38 g/L±1.19 g/L,明显低于对照细胞。结论 HBV能够在细胞水平抑制Apo E的合成和分泌。

Objective To explore the effect of hepatitis B virus（HBV） on the expression of apolipoprotein E（Apo E） at the celluar level.Methods The difference of Apo E mRNA expression between Hep G2.2.15 cell（with the whole HBV genome integrated） and the control cell Hep G2 was measured by reverse transcriptase polymerase chain reaction（RT-PCR）;HBV infectious clone p HBV1.3 was transfected into Hep G2 cell by Lipofectamine 2000 transfection reagent,and Hep G2 cell transfected with an empty vector p Blue-ks as used a control,the difference between Apo E mRNA expression the two cells was measured by RT-PCR,and Apo E levels in the supernatant of Hep G2 cell were measured by biochemical analyzer Olympus AU5400.Results The expression of Apo E mRNA was much lower in Hep G2.2.15 cell integrated with the whole HBV genome than that in the control cell Hep G2.As compared with Hep G2 cell transfected with p Blue-ks,the expression of Apo E mRNA was significantly reduced,and the Apo E level in the supernatant of Hep G2 transfected with p HBV1.3 was much lower than the control（3.38 ± 1.19 g/L vs 4.45 ± 1.62 g/L）.Conclusion HBV can down-regulate the the synthesis and secretion of Apo E at the celluar level.