肠道病毒71型感染人脑微血管内皮细胞的初步研究  

Enterovirus 71 infecting human brain microvascular endothelial cells:apreliminary study

作　　者：罗文英[1,2] 赵卫[3] 彭亮[4] 余晶仪[1] 张立科[1] 张文炳[1] 曹虹[1] 

机构地区：[1]南方医科大学公共卫生与热带医学学院微生物学系,广州510515 [2]广东医学院附属医院检验科,湛江524001 [3]南方医科大学公共卫生与热带医学学院P3实验室,广州510515 [4]广州医科大学第二附属医院检验科,广州510260

出　　处：《第二军医大学学报》2014年第3期240-245,共6页Academic Journal of Second Military Medical University

基　　金：国家自然科学基金(81301478);南方医科大学公共卫生与热带医学学院院长基金(GW201201);广东医学院博士启动课题(XB1325)~~

摘　　要：目的 初步探讨肠道病毒71型（enterovirus 71,EV71）穿血脑屏障（blood-brain barrier,BBB）的相关机制.方法 用人脑微血管内皮细胞（human brain microvascular endothelial cells,HBMECs）建立体外BBB模型,采用EV71毒株感染HBMECs,并设置阴性对照.通过形态学观察、免疫荧光、透射电镜及实时荧光定量PCR方法探讨EV71能否感染HBMECs,激光共聚焦方法观察EV71能否诱导HBMECs微丝骨架的重排.结果 EV71感染后能诱导HBMECs的细胞病变,细胞内可见EV71抗原红色荧光表达及直径为20～30 nm的EV71颗粒存在,实时荧光定量PCR方法证实EV71能在HBMECs内复制,不同时间水平差异有统计学意义（P＜0.01）.细胞感染EV71后失去正常形态,变得极不规则,胞内微丝排列紊乱,极性消失,微丝聚集在细胞边缘.结论 本研究首次证明EV71可以感染HBMECs并复制,并且可以诱导部分HBMECs的骨架重排.Objective To explore the mechanism by which enterovirus 71(EV71)crossing the blood-brain barrier(BBB).Methods BBB models were established by human brain microvascular endothelial cells(HBMECs)in vitro and were randomly assigned to control and EV71-infected groups.The cytotoxic effect of EV71on HBMECs was examined by morphological observation,immunoflurescence,transmission electron microscope and real-time PCR.In addition,the cytoskeletal alterations in HBMECs were observed by laser confocal method.Results EV71infection could lead to cytopathic changes of HBMECs.Immunofluorescent staining confirmed the presence of EV71antigen(red fluorescence)and 20-30nm EV71particles in EV71-infected HBMECs,but not in mock infected cells.Viral replication in HBMECs was shown by realtime PCR in a time-dependent manner(P<0.01).EV71infected cell lost the normal morphology and polarity,with disarranged actin filaments.Conclusion For the first time we prove that EV71can infect HBMECs and can replicate in the cells,and can induce cytoskeleton changes in some HBMECs.

关 键 词：肠道病毒71型 血脑屏障 人脑微血管内皮细胞 细胞骨架 

分 类 号：R373.25[医药卫生—病原生物学]