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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。
Objective To detect the expression of HBV markers in human chorion trophoblast cells after HBV infection and the morphological changes of cells, and to explore the relationship between HBV infection of trophoblast cells and HBV intrauterine transmission. Methods HBV infected sera were incubated with primary cultured human choriotome trophoblast cells and passage cells JEG-3 for 8 to 48 hours. The morphological changes and intercellular junctions were observed by inverted microscope. Trophoblast cells were detected by immunofluorescence The HBsAg expression was detected by real-time quantitative PCR and the apoptosis of trophoblast cells was detected by TUNEL. Results Trophoblast cells incubated with HBV-infected sera showed no significant changes in the morphology and intercellular junctions of trophoblast cells. However, the positive expression of HBsAg in trophoblast cells was detected by immunofluorescence and detected by fluorescence quantitative PCR Trophoblastic cells in the presence of HBVDNA; the same time, apoptosis test results show that incubated with HBV-infected serum, trophoblast cells significantly increased the number of apoptosis. Conclusions HBV can infect trophoblast cells cultured in vitro. HBV infection can induce trophoblast apoptosis and the infection and apoptosis of trophoblast cells may be related to the intrauterine transmission mechanism of HBV.