空间诱变菌感染对模拟失重RAW264.7巨噬细胞miR-1224-5p表达的影响及生物信息学分析  

Expression and Bioinformatics Analysis of miR-1224-5p in RAW264.7 Macrophages Infected by Space Mutation E.coli under Simulated Weightlessness

作　　者：刘蓉[1,2] 薛光泽 袁敏 王静宇[3] 王俊锋[4] 程江 袁明[3] Liu Rong;Xue Guangze;Yuan Min;WangJingyu;Wang Junfeng;Cheng Jiang;Yuan Ming(不详;State Key Laboratory of Space MedicineFundamentals and Application,China Astronaut Research and Training Center,Beijing 100094,Chi-na)

机构地区：[1]中山大学附属第七医院,广东深圳518107 [2]石河子大学第一附属医院检验科,新疆石河子832000 [3]中国航天员科研训练中心航天医学基础与应用国家重点实验室,北京100094 [4]中国人民解放军总医院第二医学中心呼吸科,北京100853

出　　处：《航天医学与医学工程》2020年第5期384-389,共6页Space Medicine & Medical Engineering

基　　金：国家重点基础研究发展计划(2014CB744404);军队后勤科研重点项目(BWS17J030)。

摘　　要：目的探讨模拟失重下空间诱变大肠杆菌T1-13感染对RAW264.7巨噬细胞miR-1224-5p表达的影响及其生物学意义。方法采用细胞回转模拟失重,RAW264.7巨噬细胞随机分为对照组(Con)、对照染菌组(Con+T1-13)、模拟失重组(SW)及模拟失重染菌组(SW+T1-13),模拟失重细胞置于回转器回转培养72h,染菌细胞以T1-13刺激6h诱导炎症反应。采用qRT-PCR技术检测各组细胞内miR-1224-5p表达,对其靶基因进行预测并开展Gene Ontology(GO)功能富集、KEGG信号通路分析和STRING蛋白互作分析。结果 qRT-PCR结果表明,与Con组相比,SW+T1-13组RAW264.7巨噬细胞中miR-1224-5p表达显著下调(P<0.05)。生物信息学分析显示,miR-1224-5p靶基因可显著富集于轴突导向、PI3K-AKT、FOXO、Notch、Wnt及MAPK等信号通路;蛋白互作网络中Kras、MAPK8、IL-6、MAPK14、Myc、Stat1、Pik3r1等蛋白处于网络中心位置。结论模拟失重下空间诱变大肠杆菌T1-13感染可导致RAW264.7巨噬细胞中miR-1224-5p表达显著下调,miR-1224-5p可能通过其靶基因参与了炎症相关的分子调控。Objective To explore the effect of space mutation E.coli T1-13 infection on miR-1224-5p expression in RAW264.7 macrophages under simulated weightlessness and its biological significance.Methods Simulated weightlessness was induced by clinostat,RAW264.7 macrophages were randomly divided into control group(Con),control macrophages infected by Ec.oli T1-13(Con+T1-13),simulated weightlessness(SW),and simulated weightlessness macrophages infected by E.coli T1-13(SW+T1-13)groups.The simulated weightlessness cells were placed on a gyrator for 72 h.The infected cells were stimulated with space mutation E.coli T1-13 for 6 hours to induce inflammatory response.qRT-PCR technology was used to detect the expression of miR-1224-5p in each group,and the predicted target genes were analyzed by Gene Ontology(GO)enrichment,KEGG pathway and STRING protein-protein interaction analysis.Results The qRT-PCR result showed that,compared with the Con group,miR-1224-5p expression was significantly down-regulated in SW+T1-13 group(P<0.05).Bioinformatics analysis showed that the target genes of miR-1224-5p were significantly enriched in Axon guidance,PI3K-AKT,FOXO,Notch,Wnt and MAPK signaling pathways,and the proteins such as Kras,MAPK8,IL-6,MAPK14,Myc,Stat1 and Pik3 r1 were located in the central position of protein-protein interaction network.Conclusion Space mutation Ec.oli T1-13 infection under simulated microgravity can induce significant down-regulation of miR-1224-5p in RAW264.7 macrophages,and miR-1224-5p may take part in the molecular regulation of inflammatory response by its target genes.

关 键 词：模拟失重 空间诱变大肠杆菌 MIRNAS RAW264.7巨噬细胞 炎症 

分 类 号：R852.22[医药卫生—航空、航天与航海医学]