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目的研究肠道病毒71型(EV71)对小胶质细胞的活化作用。方法采用细胞病变法将EV71接种小胶质细胞(BV2),通过观察接种后不同时间的荧光强度,确定EV71对小胶质细胞的感染性,同时采用实时荧光定量PCR检测病毒基因组;应用Griess法和流式细胞术检测EV71激活小胶质细胞后一氧化氮(NO)和炎性细胞因子的分泌情况。结果荧光病毒EV71接种小胶质细胞可引起细胞形态改变,但经染细胞未观察到明显的绿色荧光;EV71接种后不同时间小胶质细胞的病毒载量未发生显著变化,但NO的合成和IL-6的表达均显著升高(48h时二者含量分别为40.9μmol/L和8 793.2pg/ml),表明小胶质细胞已被EV71激活。结论 EV71不能在小胶质细胞内进行复制,但可以激活小胶质细胞,并高表达炎症介质NO和IL-6。
Objective To study the activation of microglia by enterovirus 71 (EV71). Methods EV71 was inoculated with BV2 by cytopathic method, and the infection of microglia by EV71 was determined by observing the fluorescence intensity at different time after inoculation, and the viral genome was detected by real-time fluorescence quantitative PCR. The Griess method And flow cytometry were used to detect the secretion of nitric oxide (NO) and inflammatory cytokines after microglia EV71 activated. Results Fluorescent virus EV71 inoculated microglia could induce cell morphological changes, but no obvious green fluorescence was observed in infected cells. The viral load of microglia did not change significantly at different time after inoculation of EV71, but NO synthesis and IL-6 expression were significantly increased (48h when the content was 40.9μmol / L and 8793.2pg / ml), indicating that microglia have been activated EV71. Conclusion EV71 can not replicate in microglia, but it can activate microglial cells and overexpress inflammatory mediators NO and IL-6.