miR-136-5p调控IL-17诱导大鼠星形胶质细胞炎症反应  

MiR-136-5p regulates IL-17-induced inflammatory response in rat astrocytes

作　　者：何基琛 宗少晖[1] 曾高峰[2] 彭小明[1] 邓贵营 高云兵[1] HE Ji - chen , ZONG Shao - hui, ZENG Gao - feng , PENG Xiao - ming, DENG Gui - ying , GAO Yun - bing(Department of Spine Osteopathia, the First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi, China)

机构地区：[1]广西医科大学第一附属医院脊柱骨病外科,广西南宁530021 [2]广西医科大学公共卫生学院,广西南宁530021

出　　处：《广东医学》2018年第15期2274-2278,共5页Guangdong Medical Journal

基　　金：国家自然科学基金资助项目(编号:81560351)

摘　　要：目的探讨miR-136-5p靶向NF-κB/A20调控白细胞介素(IL)-17诱导星形胶质细胞炎症反应的研究。方法体外培养SD大鼠原代星形胶质细胞,免疫荧光染色鉴定。构建miR-136-5p过表达及表达抑制的重组慢病毒载体并转染细胞,荧光显微镜下观察并计算转染效率。Real-time PCR及抗体芯片检测IL-17刺激后各组(阴性组:转染LV-ctrl病毒、过表达组:转染LV-miR-136-5p病毒、抑制组:转染LV-miR-136-5p-inhibition病毒)大鼠星形胶质细胞炎性和趋化因子[白细胞介素-1β(IL-1β)、IL-6、肿瘤坏死因子-α(TNF-α)、巨噬细胞炎性蛋白2(MIP-2)、单核细胞趋化蛋白1(MCP-1)]的相对表达情况。Western blot检测p-NF-κB、A20蛋白的相对表达水平。结果与阴性组相比,过表达组细胞炎性和趋化因子的mRNA和蛋白表达增加,A20蛋白表达减少,p-NF-κB蛋白表达增加,同时,抑制组的测定结果与之相反。结论 miR-136-5p能够通过NF-κB/A20信号通路来促进IL-17诱导的星形胶质细胞产生上述炎性和趋化因子。Objective To investigate the effect of miR-136-5p targeting nuclear factor-κB（ NF-κB）/A20 on the inflammatory response of rat astrocytes induced by IL-17. Methods Primary astrocytes of SD rats were cultured in vitro and identified by immunofluorescence staining. MiR-136-5p overexpression and expression-inhibited recombinant lentiviral vectors were constructed and transfected into rat astrocytes. The transfection efficiency was calculated under fluorescence microscope. Real-time PCR and antibody microarray were used to assess the relative expression levels of IL-1β,IL-6,tumor necrosis factor-α（ TNF-α）,macrophage inflammatory protein-2（ MIP-2） and monocyte chemoattractant protein-1（ MCP-1） in rat astrocytes after IL-17 stimulation. The relative expression levels of p-NF-κB and A20 protein were determined by Western blot. Results Compared with the negative group,miR-136-5p was transfected into rat astrocytes and stimulated by IL-17. Overexpression of miR-136-5p increased the expression of IL-1β,IL-6,TNF-α,MIP-2 and MCP-1 mRNA and protein in cells; reduced the relative expression of A20 protein was decreased; and increased the relative expression of p-NF-κB protein. However,the results of the inhibition group were opposite to those in the over expression group. Conclusion MiR-136-5p promotes the expression of inflammation cytokines and chemokines via NF-κB/A20 signaling pathway in rat astrocytes stimulated with IL-17.

关 键 词：星形胶质细胞 miRNA NF-κB/A20信号通路 炎性因子和趋化因子 

分 类 号：R651.2[医药卫生—外科学]