gga-miR-142-5p负调控chMDA5抗传染性法氏囊病病毒感染的分子机制  

GGA-MIR-142-5P IS A NEGATIVE REGULATOR OF CHMAD5 AND PROMOTES REPLICATION OF INFECTIOUS BURSAL DISEASE VIRUS IN DT40 CELLS

作　　者：欧阳伟[1,2] 潘群兴 钱晶[1] 王晶宇[1] 王晓丽[1] 夏兴霞[1] 诸玉梅[1] 王永山[1] OUYANG Wei;PAN Qun-xing;QIAN Jing;WANG Jing-yu;WANG Xiao-li;XIA Xing-xia;ZHU Yu-mei;WANG Yong-shan(Key Laboratory of Veterinary Biological Engineering and Technology,Ministry of Agriculture,Institute of Veterinary Medicine,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China;Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses,Yangzhou 225009,China)

机构地区：[1]江苏省农业科学院兽医研究所农业部兽用生物制品工程技术重点实验室,南京210014 [2]江苏省动物重要疫病与人兽共患病防控协同创新中心,扬州225009

出　　处：《中国动物传染病学报》2018年第6期24-33,共10页Chinese Journal of Animal Infectious Diseases

基　　金：国家重点研发计划项目(2016YFD0500800;2017YFD0500706);国家自然科学基金(31572504;31772723);江苏省自然科学基金(BK20141381;BK20151366);江苏省农业科技自主创新资金项目(CX(16)1052)

摘　　要：传染性法氏囊病(infectious bursal disease,IBD)是由传染性法氏囊病病毒(Infectious bursal disease virus,IBDV)引起的一种高度危害养鸡业的传染病。chMDA5在识别病原相关分子模式(pathogen associated molecular pattern,PAMPs)及激活宿主的天然免疫应答中具有重要的作用,但chMDA5识别IBDV的分子机制仍然不清楚。本研究用IBDV感染DT40细胞,在IBDV感染的DT40细胞中,chMDA5的表达水平显著升高。chMDA5过表达可以抑制IBDV的复制,并激活IFN-β promoter和Mx promoter活性,chMDA5抑制表达则得到相反的结果;chMDA5激活IFN-β promoter活性主要是通过IRF7通路。用生物信息学方法和双荧光素酶报告系统分析发现gga-miR-142-5p可作用于chMDA5的3'UTR;qRT-PCR和免疫印迹分析表明gga-miR-142-5p可以使chMDA5的蛋白水平表达降低,但mRNA水平没有变化;gga-miR-142-5p可依赖IRF7通路使IFN-β promoter和Mx promoter活性降低,IBDV复制水平升高。本研究表明,gga-miR-142-5p可以作用于chMDA5的3'UTR负调控chMDA5的表达从而影响其下游IFN-β promoter和Mx promoter活性来发挥抗IBDV感染的作用。Chicken melanoma differentiation-associated gene 5(chMDA5)is a key pattern recognition receptors(PRRs)in recognition of RNA virus infection and initialization of an antiviral innate immune response,and microRNAs(miRNAs)are involved in the regulation of chMDA5 to sense RNA virus infection in chickens.However,it remains elusive for its roles in antiviral activity against infectious bursal disease virus(IBDV)infection and regulation of microRNAs(miRNAs)on chMDA5 expression in chicken cells.In this study,we found that the expression of chMDA5 was significantly increased in IBDV-infected-DT40 cells.Over-expression of chMDA5 activated IFN-βpromoter and Mx promoter mainly through IRF7-dependent pathway and inhibited the replication of IBDV in DT40 cells while opposite effects were observed when chMDA5 was knockdown by siRNA.In addition,we proved that gga-miR-142-5p was a regulator of chMDA5 based on bioinformatics analysis and dual-luciferase reporter system.The gga-miR-142-5p reduced the expression of chMDA5 protein,which promoted IBDV replication and decreased the activities of IFN-βand Mx promoters mainly through IRF7-dependent pathway but had no effect on NF-κB-dependent pathway in DT40 cells.In conclusion,these results suggested that gga-miR-142-5p was a negative regulator of chMDA5 and promoted IBDV replication in DT40 cells mainly through the IRF7-dependent pathway.

关 键 词：传染性法氏囊病病毒 gga-miR-142-5p chMDA5 天然免疫 

分 类 号：S852.659.4[农业科学—基础兽医学]