ISSN 1003-8280 CN 10-1522/R 中国疾病预防控制中心 主办
收稿日期: 2016-02-19
网络出版日期: 2016-06-20
The role of cell membrane cholesterol in Japanese encephalitis virus and enterovirus 71 infection of human neuronal cells
Received date: 2016-02-19
Online published: 2016-06-20
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(qRT-PCR)检测MβCD对2种病毒mRNA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mRNA表达水平的变化。结果 MβCD的最高工作浓度为5 mmol/L;qRT-PCR显示MβCD预处理神经细胞可分别抑制JEV和EV71 mRNA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mRNA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。
陈生林, 朱耐伟, 朱勇喆, 徐庆强, 彭浩然, 戚中田 . 细胞膜胆固醇在流行性乙型脑炎病毒及肠道病毒71型感染人神经细胞中的作用[J]. 中国媒介生物学及控制杂志, 2016 , 27(3) : 253 -256 . DOI: 10.11853/j.issn.1003.8280.2016.03.010
Objective To investigate the role of cell membrane cholesterol in Japanese encephalitis virus (JEV) and enterovirus 71 (EV71) infection of human neuronal cells.Methods CCK-8 cell counting assay was conducted to detect the cytotoxicity of cholesterol depletion agent, ethyl-β-cyclodextrin (MβCD) on SH-SY5Y cells. The impact of MβCD on JEV and EV71 mRNA expression was validated by quantitative real-time PCR (qRT-PCR) after SH-SY5Y was pretreated by MβCD. Then MβCD was added at different stages of virus infection and detected the expression level of JEV and EV71 mRNA. Results The highest working concentration of MβCD was 5 mmol/L.Data of qRT-PCR showed that the expression of JEV and EV71 mRNA was inhibited by pretreatment of MβCD (P < 0.05), while addition of MβCD at 60 min after the incubation of EV71 had little effect (P > 0.05).Conclusion Cell membrane cholesterol plays an important role in JEV and EV71 infection of human neuronal cells, but acts at different stages.
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