巨细胞病毒在体外培养血管内皮细胞中的增殖及对细胞形态和ATP酶活性的影响.pdfVIP

· 32· J Chin Microcirc Feb．2008，Vo1．12，No．1 文章编号：1007—8568(2008)O1-0032-04 · 临床研究 · 巨细胞病毒在体外培养血管内皮细胞中的 增殖及对细胞形态和ATP酶活性的影响 姚 磊，何作云 【摘要】 目的 探讨巨细胞病毒(HCMV)在体外培养血管内皮细胞中增殖及对细胞形态和ATP 酶活性的影响。方法 应用HCMV-AD169株感染体外培养的脐静脉血管内皮细胞，应用TCID 方法测 定HCMV在ECs中的增殖变化，荧光定量PCR方法测定培养液DNA量的变化，运用化学方法测定感 染总ATP酶活性的变化，电子显微镜观察感染细胞的形态改变。结果 HCMV感染后12 h即可检测 到病毒，以后病毒滴度逐渐上升，48 h病毒增殖达高峰，随后缓慢下降。培养液HCMV—DNA量也呈现 相似的变化。感染后48 h后细胞胞浆可见微泡样结构。电镜结果显示48 h后可见线粒体肿胀、内质 网扩张、溶酶体和高尔基体增生，次级溶酶体增多，病毒样颗粒及细颗粒病毒样包涵体。感染后24 h细 胞总ATP酶开始下降，48、72 h与对照组相比下降明显(P0．01)。结论 HCMV在体外对ECs有一 定程度的直接作用，这可能作为启动因素之一在HCMV致动脉粥样硬化的发病机制中发挥重要作用。 【关键词】 巨细胞病毒；内皮细胞；动脉粥样硬化；直接作用 中图分类号：Q25 文献标识码：B The Multiplication and Morphology Changes of ECs After Human Cytomegalovirus Infection in Vitro YAO Lei，HE Zuo-yun．Department of Clinical Laboratory of Xinqiao Hospital Third，Military Medical University Chongqing 40037，China 【Abstract】Objective To study the multiplication and morphology changes of ECs after human cyto- megalovirus infection in vitro．Methods We used HCMV AD169 strain infected ECs in vitro and built the model of HCMV infection to cel1．At different time after infection(12，24，48，72h)，the multiplication changes of HCMV were observed；the cytopathic effect of ECs was studied by light microscope and electron microscope， the ATPase activity was detected by chemical methods．Results HCMV could replicate in ECs in vitro，the virus titer and HCMV DNA copies changed with the infection time，and at 48h after infection，the titer of HC— MV reached the peak．the copies of HCMV—DNA were 13567．7±1248．6 copies／ml but later showed decrea— sing yields of virus．Compared with the control group，the total ATPase activity became lower after HCMV in— fection．The H．E stain and electron microscope results showed some morphology changes of ECs after HCMV infection．Conclusion The study demonstrates that th