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目的探讨细胞膜胆固醇在流行性乙型脑炎病毒(JEV)和肠道病毒71型(EV71)感染人神经细胞中的作用。方法 CCK-8法检测胆固醇删除剂甲基-β-环糊精(MβCD)对人神经母细胞瘤SH-SY5Y细胞的毒性。分别用JEV和EV71感染MβCD预处理的SH-SY5Y细胞,实时荧光定量PCR(q RT-PCR)检测MβCD对2种病毒m RNA表达的影响。在病毒感染细胞的不同时间加入MβCD,观察2种病毒m RNA表达水平的变化。结果 MβCD的最高工作浓度为5 mmol/L;q RT-PCR显示MβCD预处理神经细胞可分别抑制JEV和EV71 m RNA的表达(P<0.05);MβCD在EV71感染细胞后60 min加入则无法抑制病毒m RNA表达(P>0.05)。结论细胞膜胆固醇在JEV和EV71感染人神经细胞中均发挥作用,但作用时间不一致。
Objective To investigate the role of cell membrane cholesterol in the inactivation of human neural cells by Japanese encephalitis virus (JEV) and enterovirus 71 (EV71). Methods CCK-8 method was used to detect the toxicity of cholesterol-deleted methyl β-cyclodextrin (MβCD) on human neuroblastoma SH-SY5Y cells. The SH-SY5Y cells pretreated with MβCD were infected with JEV and EV71, respectively. The effect of MβCD on the expression of m RNA in the two viruses was detected by real-time fluorescence quantitative PCR (q RT-PCR). MβCD was added into the infected cells at different time points to observe the changes of m RNA expression level of the two viruses. Results The maximal working concentration of MβCD was 5 mmol / L. The expression of JEV and EV71 m RNA was inhibited by MβCD pretreatment by qRT-PCR (p <0.05). MβCD could not be inhibited by 60 min after EV71 infection Virus m RNA expression (P> 0.05). Conclusion Cell membrane cholesterol plays a role in both JEV and EV71 infection of human neural cells, but the action time is inconsistent.