MiR-125b-1-3p在轮状病毒复制中的作用研究  

Research on the role of miR-125b-1-3p in rotavirus replication

作　　者：纳璐 耿盼盼 周艳[1] 姚石宝[1] 郭梅 李鸿钧[1] Na Lu;Geng Panpan;Zhou Yan;Yao Shibao;Guo Mei;Li Hongjun(Molecular Biological Chamber,Institute of Medical Biology,Chinese Academy of Medical Science,Peking Union Medical College,Kunming 650031,China;Laboratory of Experimental Hematology and Biochemistry,Institute of Radiation Medicine,Military Medical Research Institute,Beijing 100850,China)

机构地区：[1]中国医学科学院,北京协和医学院医学生物学研究所分子生物室,昆明650031 [2]军事医学研究院辐射医学研究所实验血液学与生物化学研究室,北京100850

出　　处：《中华微生物学和免疫学杂志》2021年第6期433-439,共7页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金(31700154);云南省重大科技专项(生物医药)(2018ZF006);云南省应用基础研究计划面上项目(2019FB020)。

摘　　要：目的研究miR-125b-1-3p在轮状病毒(rotavirus)复制过程中的作用和调控机制。方法将MA104细胞中的miR-125b-1-3p分别上调和下调后感染轮状病毒,通过RT-PCR分析miR-125b-1-3p的表达情况和轮状病毒拷贝数;免疫荧光分析miR-125b-1-3p对轮状病毒蛋白表达情况的影响;Western blot分析miR-125b-1-3p参与调控的相关蛋白表达情况。结果轮状病毒感染细胞后,miR-125b-1-3p的表达水平明显上调,上调miR-125b-1-3p后,轮状病毒的VP7、NSP3基因拷贝数下降,下调miR-125b-1-3p后,轮状病毒的VP7、NSP3基因拷贝数明显升高;上调miR-125b-1-3p的表达水平后轮状病毒蛋白荧光数下降,下调miR-125b-1-3p后,病毒蛋白荧光数增加;轮状病毒感染细胞16 h后PI3K/Akt通路活性受到抑制,上调miR-125b-1-3p能够抑制PI3K/Akt通路的活化。结论miR-125b-1-3p通过调控PI3K/Akt通路抑制轮状病毒的复制。研究结果为探讨miR-125b-1-3p和PI3K/Akt通路之间的具体调控机制研究提供了实验基础,为轮状病毒抗感染治疗提供了靶点。Objective To investigate the role and regulatory mechanism of miR-125b-1-3p in rotavirus replication.Methods MA104 cells were infected with rotavirus after upregulation and down-regulation of miR-125b-1-3p,respectively.The expression of miR-125b-1-3p and the copy number of rotavirus were analyzed by RT-PCR.The effect of miR-125b-1-3p on the protein expression of rotavirus was analyzed by immunofluorescence.The expression of related proteins involved in the regulation of miR-125b-1-3p was analyzed by Western blot analysis.Results After rotavirus infection,the expression level of miR-125b-1-3p was significantly up-regulated,the copy number of VP7 and NSP3 gene of rotavirus decreased after up-regulation of miR-125b-1-3p,and the copy number of VP7 and NSP3 gene of rotavirus was significantly increased after down-regulation of miR-125b-1-3p.The fluorescence number of rotavirus protein decreased after upregulation of miR-125b-1-3p expression level,and increased after down-regulation of miR-125b-1-3p expression level.The activity of PI3K/Akt pathway was inhibited 16 h after rotavirus infection,and the up-regulation of miR-125b-1-3p could inhibit the activation of PI3K/Akt pathway.Conclusions MiR-125b-1-3p inhibits rotavirus replication by regulating the PI3K/Akt pathway.These results provide an experimental basis for exploring the specific regulatory mechanism between miR-125b-1-3p and PI3K/Akt pathway,and provide a target for anti-infection therapy of rotavirus.

关 键 词：轮状病毒 miRNA PI3K/AKT 病毒复制 抗病毒反应 

分 类 号：R373.2[医药卫生—病原生物学]