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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 investigate the effect of hepatitis B virus (HBV) on the expression of apolipoprotein E (Apo E) at the cellular level. Methods Reverse transcription polymerase chain reaction (RT-PCR) was used to detect the difference of Apo E mRNA expression in Hep G2.2.15 cells (HBV whole genome) and its control Hep G2 cells. Lipofectin Lipofectin2000 transfected Hep G2 cells with HBV infectious clone p HBV1.3 and established Hep G2 cells transfected with empty vector p Blue-ks as blank control. After 48 h of transfection, Apo E mRNA The level of Apo E in Hep G2 cells was detected by Olympus AU5400 automatic biochemical analyzer. Results The expression level of Apo E mRNA in Hep G2.2.15 cells transfected with HBV whole genome was significantly lower than that in Hep G2 cells transfected with pBlue-ks. Compared with Hep G2 cells transfected with pBlue-ks, Hep G2 The expression of Apo E mRNA in G2 cells was significantly decreased, while the content of Apo E in Hep G2 cells transfected with p Blue-ks was 4.45 g / L ± 1.62 g / L, while those transfected with p HBV 1.3 The content of Apo E in Hep G2 supernatant was 3.38 g / L ± 1.19 g / L, which was significantly lower than that of control cells. Conclusion HBV inhibits the synthesis and secretion of Apo E at the cellular level.