HSP27在脑心肌炎病毒体外增殖中的多重调控作用  

Multiple regulatory effects of HSP27 on the proliferation of encephalomyocarditis virus in vitro

作　　者：李向茸[1,2] 吴贝 高铭 李殿玉 李洪珊 牛宇辉 马瑞仙 张梦月 赵旭 谢晶莹 冯若飞[1,2] LI Xiangrong;WU Bei;GAO Ming;LI Dianyu;LI Hongshan;NIU Yuhui;MA Ruixian;ZHANG Mengyue;ZHAO Xu;XIE Jingying;FENG Ruofei(Key Laboratory of Biotechnology&Bioengineering of State Ethnic Affairs Commission,Biomedical Research Center,Northwest Minzu University,Lanzhou 730030,Gansu,China;Gansu Tech Innovation Center of Animal Cell,Biomedical Research Center,Northwest Minzu University,Lanzhou 730030,Gansu,China;College of Life Science and Engineering,Northwest Minzu University,Lanzhou 730030,Gansu,China)

机构地区：[1]西北民族大学生物医学研究中心、生物工程与技术国家民委重点实验室,甘肃兰州730030 [2]西北民族大学生物医学研究中心、甘肃省动物细胞技术创新中心,甘肃兰州730030 [3]西北民族大学生命科学与工程学院,甘肃兰州730030

出　　处：《微生物学报》2023年第1期333-345,共13页Acta Microbiologica Sinica

基　　金：甘肃省高等学校创新基金项目(2020B-064);西北民族大学国家级大学生创新创业训练计划(202210742022);西北民族大学本科生创新项目(X202210742293);甘肃省青年科技基金计划(20JR5RA501);西北民族大学中央高校基本科研业务费专项资金(31920210051,31920220068)。

摘　　要：【目的】探究热休克蛋白27(heat shock protein 27,HSP27)在脑心肌炎病毒(encephalomyocarditis virus,EMCV)体外增殖过程中对多种信号通路的调控作用,为明确其他宿主因子体外调控EMCV增殖的机制研究奠定基础,并为进一步揭示EMCV致病的分子机制提供有力证据。【方法】利用实时荧光定量PCR(real-time quantitative PCR,RT-qPCR)、免疫印迹、间接免疫荧光和核质分离等方法检测干扰或过表达HSP27对EMCV感染引起的细胞自噬、翻译起始因子2α(eukaryotic translation initiation factor 2αsubunit 1,EIF2S1)-应激特异性转录因子4(activating transcription factor 4,ATF4)信号通路及髓样分化因子(myeloid differentiation factor 88,MyD88)/核转录因子κB(nuclear transcription factor kappa B,NF-κB)信号通路的影响。【结果】干扰宿主细胞中HSP27的表达可促进自噬及EIF2S1-ATF4信号通路,抑制EMCV诱导的MyD88/NF-κB信号通路的活化。相反,过表达HSP27不仅能有效降低EMCV诱导的自噬及EIF2S1-ATF4信号通路,还能促进MyD88/NF-κB信号通路分子的表达、p65的核移位和炎症相关因子的转录表达。【结论】HSP27既可以通过抑制EIF2S1-ATF4信号通路来负调控EMCV诱导的自噬,还可以正向调控EMCV触发的MyD88/NF-κB信号通路,表明HSP27在EMCV感染中具有多重调控作用。[Objective]We explored the regulatory effects of heat shock protein 27(HSP27)on various signaling pathways during the proliferation of encephalomyocarditis virus(EMCV)in vitro,aiming to lay a foundation for deciphering the mechanism of other host factors in regulating EMCV proliferation in vitro and provide evidence for comprehensively revealing the pathogenic mechanism of EMCV.[Methods]Real-time quantitative PCR(RT-qPCR),Western blotting,indirect immunofluorescence,and nucleocytoplasmic separation were employed to evaluate the effects of the knockdown and overexpression of HSP27 on the EMCV infection-induced autophagy,eukaryotic translation initiation factor 2αsubunit 1(EIF2S1)-activating transcription factor 4(ATF4)signaling pathway,and myeloid differentiation factor 88(MyD88)/nuclear transcription factor kappa B(NF-κB)signaling pathway.[Results]Interfering with the expression of HSP27 in host cells promoted autophagy and activated EIF2S1-ATF4 signaling pathway,and inhibited EMCV-induced activation of MyD88/NF-κB signaling pathway.On the contrary,overexpression of HSP27 not only reduced EMCV-induced autophagy and inhibited EIF2S1-ATF4 signaling pathway,but also up-regulated the expression of the proteins in MyD88/NF-κB signaling pathway and promoted the nuclear translocation of p65 and the transcription of inflammation-related cytokines.[Conclusion]HSP27 can not only negatively regulate EMCV-induced autophagy by inhibiting the EIF2S1-ATF4 signaling pathway,but also positively regulate the EMCV-triggered MyD88/NF-κB signaling pathway,playing multiple regulatory roles in EMCV infection.

关 键 词：热休克蛋白27 脑心肌炎病毒 细胞自噬 内质网应激反应 NF-ΚB信号通路 

分 类 号：R373[医药卫生—病原生物学]