感染乙脑病毒小鼠脑组织的转录组变化分析

Analysis of transcriptome changes in brain tissue of mice with Japanese encephalitis virus infection

为探究乙型脑炎病毒对小鼠脑组织损伤相关基因的变化,本试验采用P3株乙脑病毒感染6周龄BALB/c鼠,取对照小鼠和病毒感染后发病小鼠的脑组织,筛选差异表达基因,并用GO和KEGG数据库进行分析。组织病理学观察见神经元变性坏死,血管周炎性细胞增多及胶质细胞增生;免疫组织化学染色显示大脑皮层神经元细胞内JEV抗原阳性信号显著;脑组织提取RNA后,使用Illumina HiSeq^(TM)平台进行测序,筛选出1940个差异表达基因,其中表达上调基因1813个,下调基因127个。GO富集分析显示,差异基因大多与免疫、炎症的调控等途径有关。KEGG富集分析发现,参与免疫过程的差异基因数量最多且富集最显著。显著富集途径有JAK-STAT信号通路、Toll样受体信号通路、NF-κB信号通路、细胞因子-细胞因子受体相互作用和TNF信号通路。选取8个差异基因,采用qRT-PCR进行验证,证实和转录组分析结果一致。本研究获得的小鼠感染乙脑病毒后脑的转录组数据,将有助于进一步研究乙脑的发病机制。

In order to explore the effect of Japanese encephalitis virus on mouse brain tissue damage-related genes,Japanese encephalitis virus P3strain was used to infect 6wBALB/c mice.The brain tissues of control mice and mice infected with the virus were selected to screen the differential expression genes,and the data were analyzed with GO and KEGG databases.Histopathological observations showed neuronal cell degeneration and necrosis,the number of perivascular inflammatory cells increased and glial cell hyperplasia;immunohistochemical staining showed that cerebral cortex neurons were positive for JEV antigen.RNA was extracted from brain tissue and sequenced using the Illumina HiSeq^(TM) platform,and 1940differentially expressed genes were screened,of which 1813were up-regulated and 127were down-regulated.GO enrichment analysis showed that most of the differential genes were related to immune,inflammation and regulation pathways.KEGG enrichment analysis found that the number of differential genes involved in the immunization process was the largest and the most significant enrichment.The main differentially expressed genes were enriched in the Jak-STAT signaling pathway,Toll-like receptor signaling pathway,NF-κB signaling pathway,cytokine-cytokine-receptor interaction and TNF signaling pathway.Eight differential genes were selected and tested by qRT-PCR,it further confirmed that they were consistent with transcriptome analysis.The transcriptome data of the brains of mice infected with Japanese encephalitis virus obtained in this study will help to further study the pathogenesis of Japanese encephalitis.