HBV阳性血清对胎盘滋养层细胞的影响研究

目的探讨乙型肝炎病毒(HBV)阳性血清对体外培养胎盘滋养层细胞的影响。方法体外培养胎盘滋养层细胞(JEG-3),应用HBV阳性血清处理,分别于处理后24 h、48 h和72 h采用免疫组织化学SP法检测细胞中HBsAg蛋白的表达,PCR法检测细胞中HBV DNA,annexinⅤ/PI双标法检测细胞凋亡和坏死,透射电镜观察细胞超微结构的变化并寻找病毒颗粒。结果 HBV阳性血清处理后24～72 h,JEG-3细胞中HBsAg和HBV DNA呈阳性;处理后48 h,细胞坏死率显著增加(P<0.01),凋亡率无明显变化(P>0.05);细胞超微结构显示,JEG-3胞浆内出现较多空泡状结构,溶酶体明显增多,并可见病毒样颗粒。结论 HBV阳性血清可导致体外培养胎盘滋养层细胞的HBV感染,引起胎盘滋养层细胞损伤。

Effects of HBV-positive serum on placental trophoblasts in vitro

Objective To explore the effect of HBV-positive serum on placental trophoblasts. Methods Placental trophoblast JEG-3 cells were treated with HBV-positive or negative serum. 24,48 or 72 h after treatment, HBsAg on JEG-3 cells was determined by immunohistochemistry using SP staining, and HBV DNA in the JEG-3 cells was detected by PCR. Apoptosis and necrosis of the JEG-3 cells were analyzed by annexin V/PI staining and flow cytometry. Transmission electron microscopy was used to observe the cell ultrastructure. Results 24 to 72 h after the treatment, both HBsAg and HBV DNA in JEG-3 cells were positive. In addition, 48 h after treatment, the necrosis rate of JEG-3 cells treated with HBV-positive serum was significantly increased than that in the control cells, but the apoptosis rate was not significantly changed. Uhrastructural changes of the JEG-3 ceils were seen： numerous vacuoles appeared in cytoplasm and lysosomes increased significantly, moreover, virus-like particles were found in the cytoplasm. Conclusions Placental trophoblasts treated with HBV-positive serum can be infected by HBV. HBV- positive serum may cause significant increase of cell vacuolization, marked increase of lysosomes and cell necrosis.