上海交通大学学报（医学版）

上海交通大学学报（医学版） ›› 2019, Vol. 39 ›› Issue (8): 834-.doi: 10.3969/j.issn.1674-8115.2019.08.005

• 论著·基础研究 • 上一篇    下一篇

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

金书欣 1, 2，吴婷 3，蔡飞扬 1, 2，雷蕴轩 1, 2，席晔斌 1，陈广洁 1   

  1. 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
  • 出版日期:2019-08-28 发布日期:2019-09-23
  • 通讯作者: 陈广洁，电子信箱：guangjie_chen@163.com。
  • 作者简介:金书欣（1994—），女，技术员，硕士；电子信箱： jinshuxin888@163.com。
  • 基金资助:
    国家自然科学基金（81771731）

Effect of miR-322-5p on inhibiting Th17 differentiationtargeting Akt3 in experimental autoimmune encephalomyelitis interferedinterferon-β

JIN Shu-xin1, 2, WU Ting3, CAI Fei-yang1, 2, LEI Yun-xuan1, 2, XI Ye-bin1, CHEN Guang-jie1   

  1. 1. Department of Immunology and Microbiology, Shanghai Jiao Tong University College of Basic Medical Sciences, Shanghai 200025, China; 2. Shanghai Institute of Immunology, Shanghai 200025, China; 3. Embryo Laboratory of Reproductive Medicine, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China
  • Online:2019-08-28 Published:2019-09-23
  • Supported by:
    National Natural Science Foundation of China, 81771731

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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。

关键词: 实验性自身免疫性脑脊髓炎, 干扰素 &, beta, miRNA, Th17分化

Abstract:

Objective · To investigate the effect of miR-322-5p which targets Akt3 on Th17 differentiation in experimental autoimmune encephalomyelitis (EAE) interferedinterferon-β (IFN-β). Methods · The effect of IFN-β on EAE mice which were randomly divided into IFN-β group and PBS group was examine. The percents of Th17 in the two groups were comparedfluorescence activated cell sorting. The miRNAarray 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 of Akt3 was detected after IFN-β intervention and miR-322-5p over. The target relationship between Akt3 and miR-322-5p was verifiedluciferase reporter assay. At last, the effect of Akt3 on Th17 differentiation was explored in vitro. Results · Comparedto PBSgroup, thepercent of Th17 wassignificantlydownregulated, the ofmiR-322-5p wassignificantlyupregulated and Akt3 was significantly downregulated in IFN-β group. The of Akt3 was obviously decreased after overexpressing miR-322-5p. Luciferase reporter assay showed that Akt3 was directly targetedmiR-322-5p. The percent of Th17 differentiation was greatly promotedAkt3 in vitro. Conclusion · IFN-βsignificantly ameliorates the severityof EAEbyaffecting miR-322-5p whichcan inhibitTh17 differentiationtargeting Akt3.

Key words: experimentalautoimmuneencephalomyelitis (EAE), interferon-&beta, (IFN-β), miRNA, Th17 differentiation

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