Department of Neurology 2, Meizhou People's Hospital, Meizhou, 514031, Guangdong Province, China.
Guangdong Provincial Key Laboratory of Precision Medicine and Clinical Translation Research of Hakka Population, Meizhou, 514031, Guangdong Province, China.
Microb Pathog. 2023 Sep;182:106267. doi: 10.1016/j.micpath.2023.106267. Epub 2023 Jul 22.
it was to explore the mechanism of Japanese encephalitis virus (JEV) and micro ribonucleic acid (miRNA) under high-throughput sequencing. 20 experimental mice, with good growth status and no disease infection, were selected. The cells used in the experiment included mouse microglial cell line (BV2), mouse neuroblastoma cell line (NA), and mouse brain endothelial cell line (bEnd.3). JEV titration was performed with JEV-infected cells, ribonucleic acid (RNA) in the cells was extracted, and finally the miRNA high-throughput sequencing data was analyzed. Agarose gel electrophoresis showed that the 28S and 18S electrophoresis bands were bright. Among the miRNAs detected in mouse brain tissues, 2986 were down-regulated and 1251 were up-regulated. Among miRNAs detected in NA cells, 4238 the decreasing expression and 2356 were expressed increasingly. In reducing miRNA expression, 1 multiplicity of infection (MOI) of P3 strain infection was more significant than 0.1 MOI. 10 miRNAs with significantly decreasing expression were miR-466d-3p, miR-381-3p, miR-540-3p, miR-466a-3p, miR-467a-3p, miR-574-5p, miR-199a-5p, miR-467a-5p, miR-674-5p, and miR-376b-3p. These were all obviously down-regulated in JEV-infected BV2, NA, and bEnd.3 neurons. High-throughput sequencing of JEV-infected mouse brain tissues and mouse neuronal cells found that JEV infection led to down-regulation of overall miRNA expression in host cells.
本研究旨在通过高通量测序探索日本脑炎病毒（JEV）和微小 RNA（miRNA）的作用机制。选择 20 只生长状态良好、无疾病感染的实验小鼠，实验细胞包括小鼠小胶质细胞系（BV2）、小鼠神经母细胞瘤细胞系（NA）和小鼠脑内皮细胞系（bEnd.3）。用 JEV 感染细胞进行 JEV 滴定，提取细胞中的 RNA，最后分析 miRNA 高通量测序数据。琼脂糖凝胶电泳显示 28S 和 18S 电泳带明亮。在检测到的小鼠脑组织 miRNA 中，有 2986 个下调，1251 个上调。在检测到的 NA 细胞中的 miRNA 中，有 4238 个下调表达，2356 个上调表达。在下调 miRNA 表达中，P3 株 1 感染复数（MOI）的感染比 0.1 MOI 更显著。10 个明显下调表达的 miRNA 是 miR-466d-3p、miR-381-3p、miR-540-3p、miR-466a-3p、miR-467a-3p、miR-574-5p、miR-199a-5p、miR-467a-5p、miR-674-5p 和 miR-376b-3p。这些在 JEV 感染的 BV2、NA 和 bEnd.3 神经元中均明显下调。JEV 感染小鼠脑组织和小鼠神经元细胞的高通量测序发现，JEV 感染导致宿主细胞中整体 miRNA 表达下调。
