摘要
目的研究含阻遏蛋白结构域蛋白4(arrestin domain-containing protein 4,ARRDC4)对肠道病毒71(EV71)诱导IL-6产生的调节作用和相关机制。方法利用特异性靶向ARRDC4的小干扰RNA(siRNA)在THP-1诱导分化的巨噬细胞(t-M_φ)中干扰ARRDC4的表达,无关siRNA序列干扰作为阴性对照组,采用实时定量PCR、ELISA和Western印迹等技术观察ARRDC4对IL-6产生、EV71复制以及相关天然免疫信号通路接头分子活化的影响。结果 EV71感染时,ARRDC4表达显著上调。以siRNA靶向干扰ARRDC4的表达,能抑制EV71感染诱导的IL-6 mRNA的表达和IL-6的分泌产生,促进EV71的复制。靶向干扰ARRDC4能明显抑制EV71诱导的p-65、IκBα、ERK、JNK和p38的活化。结论 ARRDC4通过增强NF-κB和MAPK信号通路的活化,促进EV71诱导的IL-6产生,从而抑制EV71的复制,正向调控EV71触发的天然免疫反应。
Objective To study the role of arrestin domain-containing protein 4（ ARRDC4） in regulation of enterovirus71（ EV71） triggered innate IL-6 production and the underlying mechanism. Methods THP-1-derived macrophages（ t-M_φ）were transfected with ARRDC4 specific siRNA and negative control siRNA. The expression and production of IL-6,replication and virus titer of EV71,and the activation of signaling pathway adaptors were analyzed with quantitative realtime PCR,ELISA and Western blot. Results Upon EV71 infection,ARRDC4 was upregulated. ARRDC4 silencing could enhance mRNA expression and production of IL-6,thus increasing the replication and virus titer of EV71. In ARRDC4 silenced t-M,the activation of p-65,IκBα,ERK,JNK and p38 was promoted. Conclusion ARRDC4 promotes EV71-t riggered IL-6 production by enhancing the activation of NF-κB and MAPK signaling pathway to inhibit EV71 infection,contributing to positive regulation of anti-EV71 innate immune responses.
作者
孟君
何雅青
杨洪
胡钢
程锦泉
MENG Jun HE Ya-qing YANG Hong HU Gang CHENG Jm-quan(School of Public Health, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China Shenzhen Center for Disease Control and Prevention, Shenzhen, Guangdong 518073, China College of Public Health, Sun Yat-Sen University, Guangzhou 510275, China)
出处
《军事医学》
CSCD
北大核心
2017年第7期576-580,共5页
Military Medical Sciences
基金
国家自然科学基金青年科学基金项目资助(81601386)
深圳市科技计划资助项目(JCYJ20160428142411948)
