人巨细胞病毒诱导ECV304血管内皮样细胞凋亡

目的 探讨人巨细胞病毒（HCMV）感染ECV304血管内皮样细胞的致病机制。方法 常规方法传代细胞和接种病毒。以PCR法和间接免疫荧光法检测病毒即刻早期基因（IE gene）及其蛋白表达；相差显微镜及电子显微镜观察病毒感染后细胞和细胞器形态变化；DNA梯度法和流式细胞术检测细胞凋亡。结果 病毒感染后72h开始出现胞体变圆、缩小、脱壁，胞质内颗粒增多，细胞边缘模糊不清的特征，可明显检测出病毒特异IEgene及其蛋白表达；电镜结果显示病毒感染96h后，胞质内明显空泡化，细胞膜微绒毛减少，核染色质浓集、边缘化，线粒体出现溶酶体化、空泡化和自噬现象，呈早期凋亡特征；DNA梯度电泳结果显示感染细胞DNA片段化特征的条带：流式细胞仪检测发现感染4d和6d时，细胞调亡率分别为4．1％和45．7％。结论 巨细胞病毒在体外细胞培养中可诱导ECV304血管内皮样细胞的凋亡。

Human cytomegalovirus induces apoptosis of ECV304 endothelial-like cells

Objective To investigate the mechanisms for the cytopathic effect(CPE) of human cytomegalovirus(HCMV) in ECV304 endothelial-like cells.Methods PCR and indirect immunofluorescence were used to detect HCMV infection by examining immediate-early(IE) gene and protein expression of the virus in ECV304 cells.Phase-contrast and electron microscopies were performed to observe the morphological changes of the infected and uninfected cells,and DNA ladder analysis and flow cytometry were carried out to study HCMV-induced cell apoptosis.Results In HCMV-infected ECV304 cells,cytopathic effects were first observed at approximately 72 h post-infection.The cells with CPE changes exhibited detachment from the monolayer,cell rounding and shrinkage.The expression of the IE gene was detected.Chromatin condensation and nuclear fragmentation along with dramatic changes of the mitochondria were observed by electron microscopy at 96 h post-infection.Cellular DNA fragmentation was observed in the infected cells,which had cells apoptotic rates of 4.1% and 45.7% at 96 h and 144 h post-infection,respectively.Conclusion HCMV can induce apoptosis of ECV304 endothelial-like cells.