3-脱氮腺苷(3-DAA)加速小鼠及仓鼠细胞日本脑炎病毒复制并下调炎症因子水平  

3-Deazaadenosine(3-DAA)accelerates Japanese encephalitis virus replication and down-regulates levels of inflammatory factors in mouse and hamster cells

作　　者：李雪云 姚敏 李越香 程治蓉 张世哲 张芳琳 吕欣 LI Xueyun;YAO Min;LI Yuexiang;CHENG Zhirong;ZHANG Shizhe;ZHANG Fanglin;LYU Xin(Department of Immunology,School of Basic Medical Sciences,Yan’an University,Yan’an 716000;Department of Microbiology and Pathogenic Biology,Basic Medical Science Academy,Air Force Medical University,Xi’an 710032,China;Company Eighteen,Regiment Five of Cadets,Basic Medical Science Academy,Air Force Medical University,Xi’an 710032,China)

机构地区：[1]延安大学医学院,陕西延安716000 [2]空军军医大学基础医学院微生物与病原生物学教研室,陕西西安710032 [3]空军军医大学基础医学院学员五大队十八队,陕西西安710032

出　　处：《细胞与分子免疫学杂志》2024年第3期235-243,共9页Chinese Journal of Cellular and Molecular Immunology

基　　金：陕西省重点研发计划项目(2021ZDLSF01-02,2020SF-227)。

摘　　要：目的利用N_(6)-甲基腺苷酸(m^(6)A)甲基化修饰抑制剂3-脱氮腺苷(3-DAA)初步明确其对日本脑炎病毒(JEV)复制的影响。方法JEV分别感染Neuro2a小鼠神经母细胞瘤细胞、N9小鼠小胶质细胞和BHK幼仓鼠肾传代细胞,然后用3-DAA处理;在C57BL/6成年小鼠足底接种JEV,腹腔注射3-DAA。实时定量PCR检测细胞与小鼠脑组织中JEV,白细胞介素1β(IL-1β)、IL-6、肿瘤坏死因子α(TNF-α)、单核细胞趋化蛋白1(MCP-1)、诱导型一氧化氮合酶(iNOS)、精氨酸酶1(Arg1)、α干扰素(IFN-α)、IFN-β、IFN-γ、CXC趋化因子配体10(CXCL10)的mRNA表达,Western blot法检测细胞及小鼠脑组织中JEV蛋白表达,观察小鼠生存情况,并通过HE染色检测小鼠脑部病理变化。结果3-DAA可剂量依赖性的促进JEV在小鼠和仓鼠细胞及小鼠脑组织的核酸复制与蛋白表达,显著加速JEV感染小鼠的发病进程,降低小鼠生存率。3-DAA的处理下调IL-6、TNF-α、CXCL10、IL-1β、iNOS多种炎症因子的表达,免疫反应减弱。结论3-DAA促进JEV感染,加速被感染细胞及小鼠死亡,提示m^(6)A修饰可能对JEV复制起负向调控作用。Objective To investigate the effect of 3-deazaadenosine(3-DAA),an N_(6)-methyladenosine(m^(6)A)methylation modification inhibitor,on the replication of the Japanese encephalitis virus(JEV).Methods Neuro2a mouse neuroblastoma cells,N9 mouse microglial cells,and BHK baby hamster kidney cells were exposed to JEV and then treated with 3-DAA.JEV was also injected into the footpad of adult C57BL/6 mice,which were then administered 3-DAA intraperitoneally.Real-time quantitative PCR was utilized to measure mRNA expression levels of JEV,interleukin 1β(IL-1β),IL-6,tumor necrosis factorα(TNF-α),monocyte chemoattractant protein 1(MCP-1),inducible nitric oxide synthase(iNOS),arginase 1(Arg1),interferon(IFN)-α,IFN-β,IFN-γ,and C-X-C motif chemokine ligand 10(CXCL10)in the cells and mouse brain tissues.Western blot analysis was used to detect JEV protein expression in the cells and mouse brain tissues.Furthermore,the survival of the mice was monitored and pathological changes in mouse brains were observed via hematoxylin and eosin(HE)staining.Results 3-DAA had a dose-dependent effect on the replication of RNA and protein expression of JEV in both BHK,N9,Neuro 2αcells and mouse brain tissues,which resulted in rapid progression of JEV infection in mice and a decrease in their survival rate.Furthermore,3-DAA suppressed the expression of inflammatory factors such as IL-6,TNF-α,CXCL10,IL-1βand iNOS,thus weakening the immune response.Conclusion 3-DAA promotes JEV infection and hastens death of infected cells and mice,indicating that m^(6)A modification may negatively regulate JEV replication.

关 键 词：3-脱氮腺苷(3-DAA) N^(6)-甲基腺苷酸(m^(6)A) 日本脑炎病毒(JEV) 复制 

分 类 号：R373.9[医药卫生—病原生物学] R512.32[医药卫生—基础医学]