摘要
目的研究乙型肝炎病毒(HBV)在人肾小球系膜细胞内的表达及对细胞凋亡的影响。方法采用脂质体转染法将C基因型重组全长乙型肝炎病毒(PHY106-CHBV)转染至人肾小球系膜细胞。收集24、48和72 h细胞培养上清液,进行HBsAg与HBeAg的定量检测;应用倒置显微镜和荧光显微镜观察细胞形态学变化;MTT法检测细胞增殖率;流式细胞仪检测细胞凋亡率。结果实验组细胞培养上清液中可检测到HBsAg和HBeAg的高表达;观察细胞形态学变化发现,转染PHY106-CHBV后可诱导人肾小球系膜细胞凋亡;MTT结果显示细胞增殖受抑;流式细胞仪检测显示细胞凋亡率增加。结论 HBV可在人肾小球系膜细胞内表达,且能抑制细胞增殖,促进细胞凋亡。
Objective To study expression of a recombinant HBV in human glomerular mesangical cells（NHMC） and its influence on cell apoptosis.Methods Recombinant full length C genotype hepatitis B virus（PHY106-CHBV） was transfected into NHMC cells by the liposome method.The supernatant of NHMC cell cultures was collected at 24,48 and 72 h after transfection.HBsAg and HBeAg were detected by the ELISA method.Morphological changes were detected by light and fluorescence microscopes after staining with Hoechst 33258.The proliferation rate and apoptosis rate of NHMC cells were separately examined using the MTT assay and flow cytometry.Results High expression levels of HBsAg and HBeAg were detected in NHMC cells after PHY106-CHBV transfection.The observed morphological changes indicated that NHMC cells′ apoptosis was induced by PHY106-CHBV.Proliferation of NHMC cells was inhibited,and the apoptotic rate was elevated as shown by flow cytometry.Conclusion HBV expression in NHMC cells led to cell proliferation arrest and promoted apoptosis.
出处
《山东大学学报（医学版）》
CAS
北大核心
2012年第1期34-38,共5页
Journal of Shandong University:Health Sciences
基金
山东省科技发展计划资助项目(2007GGWZ02054)
