乙型脑炎病毒感染小鼠原代神经元细胞的miRNA表达谱差异分析

Analysis of the miRNA Expression Profiles in the Primary Neurons of Mice Infected with Japanese Encephalitis Virus

研究乙型脑炎病毒(Japanese encephalitis virus,JEV)感染后小鼠原代神经元细胞miRNA的表达谱差异,初步探讨JEV与宿主在miRNA水平的相互作用。分别提取JEV感染48 h后的小鼠原代神经元细胞和未染毒组细胞,高通量测序分析miRNA的表达谱并进行差异分析,选取显著差异表达的miRNA进行实时定量PCR验证。结果筛选出26个差异表达显著的miRNA,其中,18个表达上调,8个表达下调。小鼠原代脑神经元细胞中mmu-miR-21a-3p、mmu-miR-223-5p、mmu-miR-147-3p、mmu-miR-155-5p和mmu-miR-146a-5p的表达促进JEV-E基因在神经元细胞中表达,而mmu-miR-301a的表达抑制JEV-E基因的表达。神经元细胞中miRNA的表达影响JEV的复制,为进一步研究JEV致神经功能异常机制提供研究方向和理论支持。

The interaction between JEV and the host at the miRNAs level was preliminarily explored by studying the miRNAs expression profiles of primary neurons in mice infected with JEV.Total RNA of JEV-infected and uninfected primary neurons of the suckling mice was extracted individually by Trizol and then analyzed miRNA expression profiles by high-throughput sequencing analysis.Significantly differentially expressed miRNAs were selected for verification by real-time quantitative PCR.Through bioinformatics analysis,26 miRNAs with significant expression differences were screened out,among which 18 miRNAs were up-regulated and 8 miRNAs were down-regulated.The results of quantitative real-time PCR of the JEV-E gene indicated that the expressions of mir-21a-3p mir-223-5p mir-147-3p mir-155-5p and mir-146a-5p could promote the expression of JEV-E gene in neurons,while the expression of mir-301a was just on the contrast.The expression of miRNAs in primary neurons could affect the replication of JEV.This study provided the theoretical basis and direction for further studies on the regulatory function of miRNAs in the mechanism of neural dysfunction induced by JEV.