知识与财富的链接
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 EV71 on 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 EV71 infection could lead to cytopathic changes of HBMECs. Immunofluorescent staining confirmed the presence of EV71 antigen (red fluorescence) and 20-30 nm EV71 particles in EV71-infected HBMECs, but not in mock infected cells. Viral replication in HBMECs was shown by real-time PCR in a time-dependent manner (P<0.01). EV71 infected cell lost the normal morphology and polarity, with disarranged actin filaments. Conclusion For the first time we prove that EV71 can infect HBMECs and can replicate in the cells, and can induce cytoskeleton changes in some HBMECs.
