畜牧兽医学报 ›› 2019, Vol. 50 ›› Issue (8): 1709-1714.doi: 10.11843/j.issn.0366-6964.2019.08.021

• 研究简报 • 上一篇    下一篇

脑心肌炎病毒感染神经细胞的基因表达谱分析

李丽敏1,2, 邹云婧1, 郝雪飘1, 赵兴华1,2, 李睿文1,2, 孙继国1,2, 袁万哲1,2*   

  1. 1. 河北农业大学动物医学院, 保定 071001;
    2. 河北省兽医生物技术工程技术研究中心, 保定 071001
  • 收稿日期:2019-01-27 出版日期:2019-08-23 发布日期:2019-08-19
  • 通讯作者: 袁万哲,主要从事预防兽医学教学与科研工作,Tel:0312-7528377,E-mail:yuanwanzhe@126.com
  • 作者简介:李丽敏(1982-),女,河北元氏人,博士,副教授,主要从事预防兽医学教学与科研工作,Tel:0312-7528372,E-mail:lilimin03@163.com
  • 基金资助:
    河北省自然基金项目(C2016204191);河北省高校百名优秀创新人才支持计划(SLRC2017039);河北省现代农业产业技术体系羊产业创新团队专项资金(HBCT2018140204)

Analysis on Gene Expression Profiling in Nerve Cells Infected by Encephalomyocarditis Virus

LI Limin1,2, ZOU Yunjing1, HAO Xuepiao1, ZHAO Xinghua1,2, LI Ruiwen1,2, SUN Jiguo1,2, YUAN Wanzhe1,2*   

  1. 1. College of Animal Medicine, Agriculture University of Hebei, Baoding 071001, China;
    2. Hebei Engineering and Technology Research Center of Veterinary Biotechnology, Baoding 071001, China
  • Received:2019-01-27 Online:2019-08-23 Published:2019-08-19

摘要: 脑心肌炎病毒(EMCV)可感染人和多种动物。为了探索EMCV对N2a细胞转录组的影响,揭示基因表达与致病性之间的关联,本研究用EMCV BD2毒株感染N2a细胞,收集感染后7 h的细胞样本,提取总RNA,使用NimbleGen杂交体系将标记好的ds-cDNA与小鼠基因表达谱芯片杂交,对芯片进行扫描,获得差异表达基因数据;应用生物信息学方法对筛查出的差异表达基因数据进行分析,筛选差异表达的免疫相关基因及信号通路;利用real-time FQ-PCR方法,对微阵列数据结果进行验证。结果表明,EMCV感染N2a细胞后共有21 143个差异表达基因;通路分析表明,差异表达的主要信号通路包括PI3K-Akt信号通路、MAPK信号通路、细胞因子间相互作用、趋化因子信号通路、TCR/RLR信号通路和细胞凋亡通路等;Real-time FQ-PCR检测的10个差异表达基因的变化情况与通过微阵列分析预测的变化一致。

关键词: 脑心肌炎病毒, 基因表达谱, N2a

Abstract: Encephalomyocarditis virus (EMCV) can infect humans and many animals. This genome-wide expression profile could contribute to exploring the association between gene expression and pathogenicity, and revealing the molecular mechanism after infection. N2a cells were infected usingEMCV BD2 strain. The cells were collected 7 hours after infection, total RNA was extracted from EMCV infected group and normal control group, respectively. Microarrays were hybridized with labeled ds-cDNA in a NimbleGen Hybridization System. Following hybridization, washing was performed. After being washed, the slides were scanned. The data of the differentially expressed genes obtained from scanning were identified and analyzed by applying bioinformatics method. Real-time FQ-PCR was performed to corroborate the results obtained with the microarray analysis. The results showed that a total of 21 143 genes were identified as differentially expressed genes at 7 hours after EMCV infection. These genes are involved in many vital functional classes, including immune response, inflammatory response, apoptosis, defense response, signal transduction. And the differentially expressed genes associated with interferon were significantly up-regulated, suggesting that interferon plays an important role in host antiviral infection. The pathway analysis demonstrated that the most significant pathways were associated with PI3K-Akt signaling pathway, MAPK signaling pathway, chemokine signaling pathway, cytokine, TLR, TCR, RLR and Jak-STAT signaling pathway. These results indicate that the host initiates a different strategy to activate immunological and inflammatory reaction to fight infection after EMCV infection. The changes of 10 differentially expressed genes detected by Real-time FQ-PCR were consistent with those predicted by microarray analysis.

Key words: encephalomyocarditis virus, gene expression profile, N2a

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