HBV的宫内传播与人绒毛膜滋养层细胞的HBV感染和凋亡关系的研究

目的检测HBV感染后人绒毛膜滋养层细胞的HBV标志物的表达和细胞的形态学变化,探讨HBV对滋养层细胞的感染与HBV宫内传播的关系。方法将HBV感染的血清与原代培养的人早孕绒毛膜滋养层细胞及传代细胞JEG-3共同孵育8～48h,通过倒置显微镜观察细胞的形态及细胞间连接,细胞免疫荧光方法检测滋养层细胞中HBsAg的表达,荧光定量PCR方法检测细胞中的HBVDNA,TUNEL方法检测滋养层细胞的凋亡。结果滋养层细胞与HBV感染的血清共同孵育后,滋养层细胞的形态及细胞间连接无明显变化;但通过免疫荧光方法检测到滋养层细胞中HBsAg的阳性表达,并通过荧光定量PCR方法检测到滋养层细胞中HBVDNA的存在;同时,细胞凋亡检测结果表明,与HBV感染的血清共同孵育后,滋养层细胞的凋亡数量明显增加。结论HBV可以感染体外培养的滋养层细胞;HBV感染可以诱导滋养层细胞凋亡,滋养层细胞的感染和凋亡可能与HBV的宫内传播机制有关。

Study on relationship between HBV intrauterine transmission and apoptosis and HBV infection of trophoblastic cells

[Objective] To investigate on the relationship between trophoblastic cells infection and intrauterine transmission of HBV, the HBV markers and cell morphology of troplastic cells after being infected with HBV were detected. [Methods] The serum containing HBV was cocultured with the primary cultured trophoblastic cells and JEG-3 cell line for 8 to 48 hours. The appearance of the cells and cell-cell conjunction were observed by inverted microscope. HBsAg in the cells was detected by cell immunofluorescence. Fluorescent quantitation-PCR was used to detect HBV DNA in the culture cells. And TUNEL technique was used to detect the cell apoptosis. [Results] After co-infection with the HBV positive serum, the appearance of the trophoblastic cells and cell-cell conjunction were not markedly changed; but the results of cell immunofluorescence showed that HBsAg could be detected in the cells. HBV DNA was detected by fluorescent quantitation-PCR. And the TUNEL results showed that the cell apoptosis increased after the co-infection. [Conclusion] In vitro cultured trophoblastic cells can be infected by HBV. Infection of HBV may induce the apoptosis of the trophoblastic cells. Apoptosis and infection of trophoblastic cells may be related with the mechanism of HBV intrauterine transmission.