乙型肝炎病毒x抗原在HBV宫内感染中的作用机制

摘要

目的 研究高复制水平HBV血清感染人绒毛膜滋养细胞株JEG-3后细胞内乙型肝炎病毒x抗原(HBxAg)及其下游蛋白在细胞内的表达情况,探讨HBxAg在HBV宫内感染中的作用.方法 对人早孕绒毛膜滋养细胞株JEG-3进行传代培养,将高复制水平乙型肝炎病毒血清(HBV-DNA定量为107～108拷贝/mL)与传代培养的JEG-3细胞共同孵育48～72 h;逆转录-聚合酶链式反应检测细胞内HBx-mRNA水平;Western blot检测细胞内HBxAg的表达;免疫组化检测细胞内HBxAg表达以及细胞内磷脂酰肌醇激酶3(PI3K)的表达;流式细胞仪检测细胞内pAKT的表达.结果 HBV感染JEG-3细胞后,在细胞中检测到HBx-mRNA,并检测到HBxAg的表达,同时发现细胞内PI3K及pAKT的表达水平明显增加,经统计学分析,差异有统计学意义(P＜0.05).结论 HBxAg/PI3K/pAKT抗细胞凋亡途径是高复制水平HBV宫内感染的可能机制.%Objective To study the expressions of hepatitis B virus X antigen (HBxAg) and downstream protein after human chorionic trophoblast cell line (JEG-3) is infected with HBV serum of high replication level and to explore the role of HBxAg in HBV intrauterine infection. Methods Human chorionic trophoblast cell line JEG-3 during early pregnancy was subcultured, and the hepatitis B virus serum of high replication level (HBV-DNA 107-108copy/ml) and subcultured cells (JEG-3) were incubated together for 48-72 hours. HBx-mRNA level was detected by reverse transcription-polymerase chain reaction) Western biot was used to detect the intracellular expression of HBxAg. Immunohistochemistry was applied to assay intracellular expression of HBxAg and the changes of the intracellular expression level of phosphatidylinositol kinase 3 (P13K); flow cytometry was used to assay the changes of intracellular expression level of pAKT. Results After human chorionic trophoblast cell line JEG-3 was infected with HBV in vitro, HBx-mRNA and HBxAg expressions were detected in the cells. And the intracellular expression levels of PI3K and pAKT significantly increased (P<0.05). Conclusion HBxAg/PI3K/ pAKT pathway is the possible mechanism of HBV intrauterine infection of high replication level.