MiR-146a高表达对神经胶质细胞BV2炎性反应的影响  

Effect of High MiR-146a Expression on the Inflammatory Reaction in BV2 Cells

作　　者：赵娜[1] 申乐[1] 姜浩武[2] 马超[2] 黄宇光[1] 

机构地区：[1]中国医学科学院北京协和医学院北京协和医院麻醉科,北京100730 [2]中国医学科学院北京协和医学院基础医学研究所人体解剖与组织胚胎学系,北京100005

出　　处：《中国医学科学院学报》2016年第1期27-32,共6页Acta Academiae Medicinae Sinicae

基　　金：国家自然科学基金(31070930;81200869)~~

摘　　要：目的观察高表达MiR-146a对细菌脂多糖(LPS)诱导后神经胶质细胞BV2炎性反应的影响。方法采用LPS刺激接受MiR-146a模拟物转染后的BV2细胞,Real-time PCR检测MiR-146a转染效率,ELISA检测促炎症因子白细胞介素-6(IL-6)和肿瘤坏死因子α(TNFα)的表达水平,Real-time PCR和Western blot法检测TLR4信号通路上肿瘤坏死因子受体相关因子6(TRAF6)和白细胞介素-1受体相关激酶1(IRAK1)的表达水平。结果与正常组比较,采用50 nmol/L的转染浓度对BV2细胞进行MiR-146a模拟物转染,可明显增加细胞内MiR-146a含量(t=5.846,P=0.0021);LPS刺激导致BV2细胞激活,可明显增加细胞内IRAK1和TRAF6表达,也可明显诱发细胞分泌更多的IL-6和TNFα;而与单纯采用LPS刺激相比,用MiR-146a模拟物转染后再接受LPS刺激作用可明显降低IL-6(t=5.200,P=0.0003)和TNFα(t=9.812,P<0.0001)的表达水平,同时明显降低细胞内TRAF6分子在基因(t=5.353,P=0.0007)和蛋白(t=6.980,P=0.0009)水平的表达,而非IRAK1的表达。结论 MiR-146a可能是通过调控TLR4信号通路中TRAF6分子的表达,负反馈抑制BV2细胞的炎性反应。Objective To explore the effect of MiR-146 a regulator function on the inflammatory response in neuroglia cell（ microglia）. Methods BV2 cells were transfected by MiR-146 a mimics,and then stimulated by lipopolysaccharide（ LPS）. MiR-146 a expression was measured by real-time polymerase chain reaction（ real-time PCR）. Interleukin（ IL）-6 and tumor necrosis factor α（ TNFα） were measured by enzymelinked immunosorbent assay（ ELISA）. Furthermore,IL-1 receptor-associated kinase 1（ IRAK1） and TNF receptor-associated factor 6（ TRAF6） were detected by PCR and Western blotting. Results Compared to the normal control group,MiR-146 a expression was significantly elevated by transfection with MiR-146 a mimics（ t =5. 846,P = 0. 0021）. The expression levels of IRAK1,TRAF6,TNFα,and IL-6 significantly increased in the LPS-stimulated BV2 cells compared to the non-stimulated BV2. The enhancement of MiR-146 a resulted insignificantly decreased IL-6（ t = 5. 200, P = 0. 0003） and TNFα（ t = 9. 812, P〈0. 0001） secretion. The mRNA（ t = 5. 353,P = 0. 0007） and protein（ t = 6. 980,P = 0. 0009） levels of TRAF6,but not IRAK1,also significantly decreased. Conclusion MiR-146 a may negatively suppress the inflammatory response of BV2 cells by regulating the expression of IRAF6 molecules in the TLR4 signaling pathway.

关 键 词：MIR-146A 小胶质细胞 神经病理性疼痛 促炎症因子 

分 类 号：R4[医药卫生—临床医学]