TRIM25通过上调IFN-β和降解病毒衣壳蛋白抑制乙型脑炎病毒在U251细胞中的复制  

TRIM25 inhibits Japanese encephalitis virus replication in U251 cells by up-regulation of the IFN-βand degrading the viral capsid protein

作　　者：陈晨 徐魁 祝壮 黄荣 冯亚岚[1] 谭宁 贺雅静 罗月 杨健[1] 袁磊 Chen Chen;Xu Kui;Zhu Zhuang;Huang Rong;Feng Yalan;Tan Ning;He Yajing;Luo Yue;Yang Jian;Yuan Lei(Institute of Basic Medicine,Institute of Basic Medicine and Forensic Medicine,North Sichuan Medical College,Nanchong 637100,China)

机构地区：[1]川北医学院,基础医学与法医学院,基础医学研究中心,南充637100

出　　处：《中华微生物学和免疫学杂志》2025年第2期99-107,共9页Chinese Journal of Microbiology and Immunology

基　　金：南充市市校合作科研项目(18SXHZ0497)。

摘　　要：目的研究三结构域蛋白25(tripartite motif-containing protein 25,TRIM25)对细胞中乙型脑炎病毒(Japanese encephalitis virus,JEV)复制的抑制作用及机制。方法JEV感染人神经胶质瘤细胞(U251细胞)和昆明种小鼠,采集细胞和小鼠脑组织样本,real-time PCR检测6种TRIM基因的转录水平,Western blot检测细胞中TRIM25蛋白的表达。构建TRIM25过表达U251细胞和A549细胞,TRIM25基因敲除U251细胞。JEV感染细胞后,用病毒蚀斑试验、real-time PCR和Western blot检测JEV的复制。real-time PCR和ELISA检测过表达TRIM25细胞IFN-β的水平。用免疫共沉淀(Co-IP)与间接免疫荧光试验研究TRIM25与病毒蛋白的相互作用。结果JEV感染上调细胞与小鼠脑组织中TRIM25的表达。TRIM25过表达抑制了JEV在U251和A549细胞中的复制,敲除TRIM25促进了JEV的复制。TRIM25过表达可上调细胞中IFN-β的水平。TRIM25与JEV的衣壳蛋白相互作用,并促使衣壳蛋白降解。结论TRIM25蛋白可以通过上调IFN-β和促使JEV衣壳蛋白降解来抑制细胞中JEV的复制。Objective To investigate the inhibitory effect of tripartite motif-containing 25(TRIM25)on the replication of Japanese encephalitis virus(JEV)in cells and its molecular mechanism.MethodsHuman glioma cells(U251 cells)and Kunming mice were infected with JEV,and then the cells and brain tissue samples were collected.The transcription levels of six TRIM genes were detected by real-time PCR,and the expression of TRIM25 in cells was detected by Western blot.U251 and A549 cells overexpressed with TRIM25 and U251 cells knocked out with TRIM25 gene were constructed.Cells were infected with JEV,and the replication of JEV was detected by viral plaque assay,real-time PCR and Western blot.The interaction of TRIM25 with viral proteins was investigated by co-immunoprecipitation(Co-IP)and indirect immunofluorescence assay.The expression of IFN-βin overexpressed TRIM25 cells was detected by real-time PCR and ELISA.ResultsJEV infection promoted the expression of TRIM25 in cells and mouse brain tissues.TRIM25 overexpression restricted JEV replication in U251 and A549 cells,while TRIM25 knockout enhanced JEV replication.TRIM25 overexpression upregulated the level of IFN-βin cells.TRIM25 interacted with JEV capsid protein and promoted the degradation of capsid protein.ConclusionTRIM25 can inhibit the replication of JEV in cells by upregulating IFN-βand promoting the degradation of JEV C protein.

关 键 词：乙型脑炎病毒 三结构域蛋白25 固有免疫 抗病毒作用 

分 类 号：S85[农业科学—兽医学]