摘要
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TLR4RNA(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.
作者
汪继
赵浩
陈秀芬
肖宗宇
朱健伟
陈文锦
张丽
徐如祥
Wang Ji;Zhao Hao;Chen Xiufen;Xiao Zongyu;Zhu Jianwei;Chen Wenjin;Zhang Li;Xu Ruxiang(Affiliated Bayi Brain Hospital,The Seventh Medical Center of the PLA General Hospital,Beijing 100700,China;Department of Neurosurgery,Affiliated Hospital of Qinghai University,Xining 810000,China;Department of Neurosurgery,Affiliated TCM Hospital of Southwest Medical University,Luzhou 646000,China)
出处
《中华神经创伤外科电子杂志》
2019年第3期163-167,共5页
Chinese Journal Of Neurotraumatic Surgery：Electronic Edition
基金
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)
