Netrin-1在伪狂犬病病毒触发的抗病毒天然免疫反应中作用的研究  

Role of Netrin-1 in pseudorabies virus infection-triggered anti-viral innate immune responses

作　　者：刘晓丹 李萍 徐志文 朱玲 

机构地区：[1]四川农业大学动物医学院动物生物技术中心,四川成都611130

出　　处：《中国兽医科学》2017年第9期1145-1149,共5页Chinese Veterinary Science

基　　金：国家级大学生创新计划项目(201410626018);四川省科技支撑计划项目(2014NZ0043)

摘　　要：为探究轴突导向生长因子Netrin-1在病毒感染中的功能,以猪伪狂犬病病毒感染MEF细胞为模型,通过荧光定量PCR检测Netrin-1基因mRNA在感染前后表达水平的变化,结果表明,伪狂犬病病毒刺激可以使得细胞下调Netrin-1 mRNA的表达水平。以小鼠Netrin-1基因保守序列设计1对特异性引物,成功将Netrin-1克隆入pcDNA3.1载体,在MEF细胞中瞬时转染重组质粒,并用伪狂犬病病毒感染细胞,针对炎症因子IL-6、干扰素IFN-β与干扰素刺激基因IFIT1和ISG15的mRNA进行qPCR验证,结果表明,Netrin-1对于干扰素与干扰素刺激基因的表达具有抑制作用。免疫印迹试验结果证明,过表达Netrin-1使得细胞核因子P65亚基表达下调。表明Netrin-1对免疫反应具有抑制作用,细胞在受到病毒感染时下调Netrin-1的表达以促进免疫反应。To investigate the function of Netrin-1 in pseudorabies infection,we evaluated the mRNA level of Netrin-1 after and before pseudorabies stimulation.The result showed that pseudorabies stimulation caused the downregulation of Netrin-1,and a flag-tag fusion protein was constructed to conduct a gain-of-function experiment,detecting of P65 by Western-blot and mRNA expression kinetics of pro-inflammatory cytokines IL-6 and IFN-β,and two interferon-stimulated genes IFIT1 and ISG15 by qPCR.The results showed that,Netrin-1 ectopic expression reversed pseudorabies-induced expression of nuclear factor subunit P65 at the protein level along with inhibition of IL-6 mRNA production.More importantly,the mRNA levels of IFN-β,IFIT1 and ISG15 were also downregulated compared with the empty vector transfection,which suggests a role in anti-viral effects elicited by downregulation of Netrin-1.

关 键 词：NETRIN-1 伪狂犬病毒 细胞核因子 干扰素 

分 类 号：S852.42[农业科学—基础兽医学]