microRNA-23b对肠道病毒71型在神经细胞中复制的抑制  

The inhibition of microRNA-23b to the replication of enterovirus 71 in neurocytes

作　　者：茅凌翔[1] 杨静[1] 沈俐[1] 吴静[2] 陈建国[3] 许化溪[2] 

机构地区：[1]镇江市疾病预防控制中心检验科,江苏镇江212001 [2]江苏大学医学院,江苏镇江212002 [3]镇江市第一人民医院检验科,江苏镇江212013

出　　处：《中国卫生检验杂志》2015年第10期1549-1551,共3页Chinese Journal of Health Laboratory Technology

基　　金：镇江市科技计划项目(SH2012052)

摘　　要：目的筛选能抑制肠道病毒71型(EV71)在人神经母细胞瘤SK-N-SH中复制的microRNA(miRNA)。方法利用miRNA芯片得到EV71感染神经细胞后的miRNA差异表达谱,将显著变化的miRNA合成miRNA模拟体和miRNA抑制体后,转染神经细胞。EV71病毒感染神经细胞后,用荧光定量RT-PCR、Western blot分别检测病毒的RNA和蛋白水平。结果 miRNA-23b模拟体转染SK-N-SH神经细胞后,与空白、阴性对照组相比,EV71导致的细胞病变效应没有显著变化;荧光定量PCR结果显示,miRNA-23b模拟体转染后,EV71病毒的RNA没有明显变化;Western blot结果显示,miRNA-23b模拟体转染后,EV71病毒VP1蛋白含量减少,并且EV71病毒滴度(TCID50)下降。结论 miRNA-23b在神经细胞中能有效抑制病毒复制,为RNA干扰技术在EV71的防治应用中奠定了基础。Objective The purpose of this study was to find some miRNA which can have antiviral activity against EV71 in neu- rocytes. Methods Differentially expressed miRNAs between the SK - N - SH cell after EV71 infection and normal cell were screened with a miRNAs chip. Some miRNAs related to EV71 infection were reversly transfected with miRNA mimics or inhibi- tors. Then, after the ceils were infected with EV71. The RNA and protein level were determined by RT - PCR and Western blot repeetively. Results miRNA -23b simulation was transfected into SK -N -SH nerve cells, and compared to the blank, nega- tive controls, the eytopathie effect caused by EV71 did not change significantly. Fluorescence quantitative PCR results showed that, after the miRNA -23b mimetibodies were transfeeted, EV71 virus RNA has no obvious change. Western blot showed, af- ter the miRNA -23b mimetibodies were transfeeted, VP1 protein content of EV71 virus reduced, and EV71 （TCID50） virus ti- ter decreased. Conclusion miRNA - 23b can effectively inhibit virus replication in nerve ceils, and RNA interference technol- ogy has laid a solid foundation＇in the prevention and treatment of EV71 application.

关 键 词：肠道病毒71型 神经细胞 微小RNA 

分 类 号：R373.25[医药卫生—病原生物学]