哺乳动物呼肠孤病毒σ3蛋白对细胞自噬的调控作用  

Study on the regulatory effect of mammalian reovirusσ3 protein on autophagy

作　　者：白辉盛 成荣荣 董平安 李骁昳 莫荣纤 李殿玉 马婷婷 李向茸[1,2,3] 冯若飞[1,2,3] BAI Huisheng;CHENG Rongrong;DONG Pingan;LI Xiaoyi;MO Rongqian;LI Dianyu;MA Tingting;LI Xiangrong;FENG Ruofei(Engineering Research Center of Key Technology and Industiliation of Cell-based Vaccine,Ministry of Education,Biomedical Research Center,Northnest Minzu University,Lanzhou 730030,China;Gansu Tech Innovation Center of A nimal Cell.Biomedical Research Ce nter,Northwest Minzu University,Lanzhou 730030,China;Key Laboratory of Biotechnology and Bioengineering of State Eithnic Affairs Cormission.Biomnedical Research Center,Northuvest Minzu University,Lanzhou 730030,China;School of Lijfe Sciences and Engineering,Northves1 Minzu University,Lanzhou 730030,China)

机构地区：[1]西北民族大学生物医学研究中心、细胞基质疫苗关键技术与产业化教育部工程研究中心,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 [4]西北民族大学生命科学与工程学院,甘肃兰州730030

出　　处：《中国兽医科学》2025年第3期291-298,共8页Chinese Veterinary Science

基　　金：西北民族大学大学生创新训练项目(X202310742266);西北民族大学中央高校基本科研业务费资金项目(31920240125-01);甘肃省高校教师创新基金项目(2025B-33)。

摘　　要：采用间接免疫荧光检测哺乳动物呼肠孤病毒(MRV)感染和σ3蛋白对自噬小体形成的影响,以及σ3蛋白与线粒体外膜转位酶20(TOMM20)的共定位情况。利用Western-blotting分析MRV感染和σ3蛋白对LC3B、SQSTM1/p62、Beclin1、ATG5等自噬标志因子及腺苷酸活化蛋白激酶(AMPK)-哺乳动物雷帕霉素靶蛋白(mTOR)信号通路因子AMPK、p-AMPK、mTOR、p-mTOR蛋白表达的影响。结果显示,MRV感染或σ3蛋白的存在使mCherry-GFP-LC3B融合蛋白由胞浆转移至自噬小体膜上,促进自噬的早期形成。同时发现MRV感染及σ3蛋白可促使自噬标志因子LC3B-Ⅰ向LC3B-Ⅱ的转变增多,上调Beclin1、ATG5、ATG7、p-AMPK的蛋白表达,并下调自噬底物SQSTM1/p62及p-mTOR的蛋白表达。本研究揭示了σ3蛋白通过AMPK/mTOR信号轴增强细胞自噬的发生,为深入阐明MRV的致病机制奠定基础,并为由MRV引起的感染性疾病的防控提供科学依据。Indirect immunofluorescence was used to detect the effects of mammalian reovirus(MRV)infection andσ3 protein on the formation of autophagosomes,and the colocalization ofσ3 protein with translocase of outer mitochondrial membrane 20(TOMM20).Western-blotting was used to analyze the ef-fects of MRV infection andσ3 protein on the expression of autophagy markers LC3B,SQSTM1/p62,Beclin 1,ATG5 and adenosine monophosphate-activated protein kinase-mammalian target of rapamycin(AMPK-mTOR)signaling pathway factors AMPK,p-AMPK,mTOR and p-mTOR.The results showed that MRV infection and the presence ofσ3 protein could transfer the mCherry-GFP-LC3B fusion protein from the cytoplasm to the autophagosome membrane,promoting the early formation of autophagy.It was also found that MRV infec-tion andσ3 protein could promote the increase of the transition from autophagy marker LC3B-Ⅰto LC3B-Ⅱ,up-regulate the protein expression of Beclin 1,ATG5,ATG7 and p-AMPK,and down-regulate the protein expressions of p-mTOR and autophagy receptor SQSTM1/p62.This study revealed thatσ3 protein enhances the occurrence of autophagy through the AMPK/mTOR signaling axis,laying a foundation for further elu-cidating the pathogenic mechanism of MRV and providing a scientific basis for the prevention and con-trol of infectious diseases caused by MRV.

关 键 词：细胞自噬 哺乳动物呼肠孤病毒 σ3蛋白 

分 类 号：S852.659.4[农业科学—基础兽医学]