摘要
目的探讨金黄色葡萄球菌（S．aureus）入侵人脐静脉内皮细胞（HUVEC）、人羊膜细胞（WISH）细胞骨架微丝、微管变化及其诱导细胞凋亡作用。方法常规方法制备S．aureus悬液，分别感染人脐静脉内皮细胞和人羊膜细胞10、30、60、120min。采用光学显微镜和荧光显微镜观察saureus感染和细胞骨架微丝、微管重排情况；采用DAPI荧光染色和DNA琼脂糖凝胶电泳分析S．aureus对HUVEC细胞诱导凋亡的影响。结果S．aureus与HUVEC和WISH细胞共育10min，即有细菌侵入，60min后，HUVEC和WISH细胞的感染率分别达（54．9±2．4）％和（56．1±2．4）％；入侵HUVEC细胞和WISH细胞10、30、60、120min时，触发微丝肌动蛋白（F—actin）重排百分率分别为（54．7±2．8）％、（63．0±2．9）％、（71．0±2．6）％、（39．5±2．7）％和（63．3±2．6）％、（65．0±2．9）％、（77．0±2．4）％、（44．0±1．8）％。10、30、60min的F-actin重排百分率明显高于120min（P〈0．05），此时微管变化不大；S．aureus感染HUVEC细胞60min时，DAPI荧光染色和DNA琼脂糖凝胶电泳均可出现细胞凋亡现象。结论S．aureus在入侵HUVEC和WISH细胞时，可能通过F—actin细胞骨架重排来完成侵袭过程，且细菌侵入60rain后可诱导HUVEC细胞出现凋亡。
Objective To investigate the changes of cytoskeleton and induced apoptosis in human umbilical venous endothelial cells （HUVEC） and WISH cells during the invasion of staphylococcus aureus. Methods S. aureus suspension was collected routinely and used to infect HUVEC and WISH cells for 10, 30, 60 and 120 min respectively. The cell-invading ability of S. aureus was observed by microscope and the rearrangement of cytoskeleton of these cells observed under fluorescent microscope. DAPI fluorescent staining and DNA agarose electrophoresis were performed to analyze the apoptosis in HUVEC ceils induced by S. aureus. Results There was bacterial invasion after staphylococcus aureus was co-incubated with HUVEC and WISH cells for 10 min. The rates of infection were （54.9± 2. 4） % and （ 56. 1± 2. 4） % at 60 min respectively. The ratios of F-actin rearrangements at 10 min, 30 min, 60 min and 120 min after the invasion of HUVEC and WISH ceils with S. aureus were different at （54. 7 ±2. 8）%, （63.0 ±2. 9）%, （71.0 ± 2.6）%, （39.5±2.7）% and （63.3±2.6）%, （65.0±2.9）%, （77.0±2.4）% and （44.0±1.8）% respectively. The ratios of F-actin rearrangements at 10 min, 30 min and 60 min were higher than those at 120 min （ P 〈 0. 05 ）. There was no change of microtubule observed in both cells at the same time. The apoptotic appearance was observed after the invasion of HUVEC cells with s. aureus at 60 min. Conclusion S. aureus may invade the HUVEC and WISH cells through F-actin rearrangement. Apoptosis is induced in HUVEC cells at 60 min.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2009年第45期3215-3219,共5页
National Medical Journal of China
基金
浙江省嘉兴市科技计划项目（2006AY2035）
