miR-322-5p靶向Akt3抑制Th17分化对干扰素β干预实验性自身免疫性脑脊髓炎的影响  

Effect of miR-322-5p on inhibiting Th17 differentiation by targeting Akt3 in experimental autoimmune encephalomyelitis interfered by interferon-β

作　　者：金书欣 吴婷[3] 蔡飞扬 雷蕴轩 席晔斌 陈广洁[1] JIN Shu-xin;WU Ting;CAI Fei-yang;LEI Yun-xuan;XI Ye-bin;CHEN Guang-jie(Department of Immunology and Microbiology,Shanghai Jiao Tong University College of Basic Medical Sciences,Shanghai 200025,China;Shanghai Institute of Immunology,Shanghai 200025,China;Embryo Laboratory of Reproductive Medicine,Renji Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200025,China)

机构地区：[1]上海交通大学基础医学院免疫学与微生物学系,上海200025 [2]上海市免疫学研究所,上海200025 [3]上海交通大学医学院附属仁济医院生殖医学科胚胎实验室,上海200127

出　　处：《上海交通大学学报（医学版）》2019年第8期834-842,共9页Journal of Shanghai Jiao tong University:Medical Science

基　　金：国家自然科学基金(81771731)~~

摘　　要：目的·探讨miR-322-5p靶向Akt3抑制Th17分化对干扰素β(interferon-β,IFN-β)干预实验性自身免疫性脑脊髓炎(experimental autoimmune encephalomyelitis,EAE)的影响。方法·建立EAE小鼠模型,设IFN-β干预组和PBS对照组。流式染色比较2组Th17的比例变化;RNA芯片检测2组小鼠miRNA的差异表达,筛选出miR-322-5p做进一步研究;软件预测miR-322-5p的靶基因为Akt3;IFN-β干预后和过表达miR-322-5p后检测Akt3的表达水平;双荧光素酶报告实验验证miR-322-5p和Akt3的直接靶向关系;体外实验观察Akt3对Th17细胞分化的影响。结果·IFN-β干预组的EAE小鼠Th17比例均显著降低,miR-322-5p的表达显著升高,而Akt3的表达明显降低;过表达miR-322-5p能显著抑制Akt3的表达,双荧光素酶报告实验显示Akt3是miR-322-5p的直接靶基因,且Akt3对Th17的体外分化有明显促进作用。结论·IFN-β可通过影响miR-322-5p靶向Akt3,进而抑制Th17分化来缓解EAE的疾病进程。Objective·To investigate the effect of miR-322-5p which targets Akt3 on Th17 differentiation in experimental autoimmune encephalomyelitis(EAE)interfered by interferon-β(IFN-β).Methods·The effect of IFN-βon EAE mice which were randomly divided into IFN-βgroup and PBS group was examine.The percents of Th17 in the two groups were compared by fluorescence activated cell sorting.The miRNA array was made to find different miRNAs between those two groups.MiR-322-5p was screened for further research.The target gene of miR-322-5p was predicted using softwares and the common predicted target gene Akt3 was got.The expression of Akt3 was detected after IFN-βintervention and miR-322-5p overexpression.The target relationship between Akt3 and miR-322-5p was verified by luciferase reporter assay.At last,the effect of Akt3 on Th17 differentiation was explored in vitro.Results·Compared to PBS group,the percent of Th17 was significantly downregulated,the expression of miR-322-5p was significantly upregulated and Akt3 was significantly downregulated in IFN-βgroup.The expression of Akt3 was obviously decreased after overexpressing miR-322-5p.Luciferase reporter assay showed that Akt3 was directly targeted by miR-322-5p.The percent of Th17 differentiation was greatly promoted by Akt3 in vitro.Conclusion·IFN-βsignificantly ameliorates the severity of EAE by affecting miR-322-5p which can inhibit Th17 differentiation by targeting Akt3.

关 键 词：实验性自身免疫性脑脊髓炎 干扰素Β MIRNA Th17分化 

分 类 号：R392.32[医药卫生—免疫学]