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中华神经创伤外科电子杂志 ›› 2019, Vol. 05 ›› Issue (03) : 163 -167. doi: 10.3877/cma.j.issn.2095-9141.2019.03.008

所属专题: 文献

基础研究

MicroRNA-182靶向抑制TLR4后减轻缺氧诱导的小胶质细胞炎症反应
汪继1, 赵浩1, 陈秀芬1, 肖宗宇2, 朱健伟3, 陈文锦1, 张丽1, 徐如祥1,()   
  1. 1. 100700 北京,解放军总医院第七医学中心附属八一脑科医院
    2. 810000 西宁,青海大学附属医院神经外科
    3. 646000 泸州,西南医科大学附属中医医院神经外科
  • 收稿日期:2018-12-04 出版日期:2019-06-15
  • 通信作者: 徐如祥
  • 基金资助:
    国家自然科学基金(81671189); 全军"十三五"军事医学创新工程项目(16CXZ001)

MicroRNA-182 targets inhibition of TLR4 and reduces hypoxia-induced microglial inflammatory response

Ji Wang1, Hao Zhao1, Xiufen Chen1, Zongyu Xiao2, Jianwei Zhu3, Wenjin Chen1, Li Zhang1, Ruxiang Xu1,()   

  1. 1. Affiliated Bayi Brain Hospital, The Seventh Medical Center of the PLA General Hospital, Beijing 100700, China
    2. Department of Neurosurgery, Affiliated Hospital of Qinghai University, Xining 810000, China
    3. Department of Neurosurgery, Affiliated TCM Hospital of Southwest Medical University, Luzhou 646000, China
  • Received:2018-12-04 Published:2019-06-15
  • Corresponding author: Ruxiang Xu
  • About author:
    Corresponding author: Xu Ruxiang, Email:
引用本文:

汪继, 赵浩, 陈秀芬, 肖宗宇, 朱健伟, 陈文锦, 张丽, 徐如祥. MicroRNA-182靶向抑制TLR4后减轻缺氧诱导的小胶质细胞炎症反应[J/OL]. 中华神经创伤外科电子杂志, 2019, 05(03): 163-167.

Ji Wang, Hao Zhao, Xiufen Chen, Zongyu Xiao, Jianwei Zhu, Wenjin Chen, Li Zhang, Ruxiang Xu. MicroRNA-182 targets inhibition of TLR4 and reduces hypoxia-induced microglial inflammatory response[J/OL]. Chinese Journal of Neurotraumatic Surgery(Electronic Edition), 2019, 05(03): 163-167.

目的

研究microRNA-182(miR-182)靶向Toll样受体4(TLR4)对小胶质细胞炎症反应的影响。

方法

利用转染试剂Lipofectamine 2000进行小胶质细胞的miR-182、小干扰TLR4 RNA(siTLR4)以及TLR4(ORF)过表达质粒的转染;通过氧糖剥夺处理,模拟缺血缺氧环境;通过荧光素酶基因检测报告检测TLR4是否是miR-182的直接结合靶点;通过Western blot、RT-PCR、ELISA检测mRNA以及蛋白的相对表达量。

结果

miR-182与TLR4在缺氧条件下小胶质细胞中的表达具有相关性,TLR4是miR-182的直接作用靶点,miR-182可以抑制TLR4的表达,同时降低小胶质细胞的炎症因子(TNF-α、IL-6、IL-1β)的释放。

结论

miR-182通过靶向抑制TLR4的表达来抑制缺氧诱导的小胶质细胞炎症反应。

Objective

To study the effect of microRNA-182 (miR-182) targeting Toll-like receptor 4 (TLR4) on microglial inflammatory response.

Methods

Microglial cells were transfected with miR-182, small interfering TLR4 RNA (siTLR4) and TLR4(ORF) overexpression plasmids using transfection reagent Lipofectamine 2000. Simulating ischemia and hypoxia by OGD treatment. Detection of TLR4 as a direct target of miR-182 by luciferase reporter assay. Detection of mRNA and relative expression of protein by RT-PCR, Western blot and ELISA.

Results

miR-182 is associated with the expression of TLR4 in microglia under hypoxic conditions, TLR4 is a direct target of miR-182, and miR-182 can inhibit the expression of TLR4, and reduce the release of inflammatory factors (TNF-α, IL-6, IL-1β) from microglia

Conclusion

miR-182 inhibits hypoxia-induced microglial inflammatory response by targeting inhibition of TLR4 expression.

图1 OGD处理不同时间后小胶质细胞miR-182、TLR4 mRNA和蛋白相对表达量
图2 荧光素酶基因测定报告结果
图3 小胶质细胞转染miR-182后TLR4的表达情况
图4 小胶质细胞转染siTLR4、TLR4过表达质粒后TLR4表达情况
图5 小胶质细胞的炎症因子表达水平
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