miR-21通过NF-κB信号通路影响血管紧张素Ⅱ诱导的肾小球系膜细胞炎症因子表达  

miR-21 affects angiotensin Ⅱ-induced inflammatory factor expression in glomerular mesangial cells via NF-κB signaling pathway

作　　者：彭君[1] 彭家清[1] 秦鹏[1] 罗先荣 张敏[1] PENG Jun;PENG Jiaqing;QIN Peng;LUO Xianrong;ZHANG Min(Department of Nephrology,Jingzhou Central Hospital in Hubei Province,Jingzhou 434020,China)

机构地区：[1]湖北省荆州市中心医院肾内科,434020

出　　处：《免疫学杂志》2020年第5期383-389,共7页Immunological Journal

基　　金：荆州市科技发展计划(2015049)。

摘　　要：目的探讨miR-21对血管紧张素Ⅱ(AngⅡ)诱导肾小球系膜细胞增殖和炎症因子的影响。方法以AngⅡ诱导体外培养人肾小球系膜细胞HMC,在AngⅡ诱导的HMC细胞中转染miR-21模拟物。采用实时荧光定量PCR检测miR-21表达水平,噻唑蓝(MTT)法检测细胞增殖能力,Western blot检测NF-κB信号通路核心分子IκBα和p65蛋白及磷酸化水平,酶联免疫吸附法(ELISA)检测炎症相关因子白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)和肿瘤坏死因子-α(TNF-α)含量。采用NF-κB信号通路激活剂PMA处理,检测对炎症因子表达的影响。结果 AngⅡ组HMC细胞中miR-21表达水平较Control组降低(P<0.05),细胞增殖能力明显升高(P<0.05),炎症因子IL-6、IL-1β和TNF-α的表达升高(P<0.05)。进一步在AngⅡ组过表达miR-21后,IκBα和p65蛋白磷酸化水平显著降低(P<0.05),细胞增殖能力明显降低(P<0.05),IL-6、IL-1β和TNF-α的表达亦降低(P<0.05)。NF-κB信号通路激活剂处理后,IL-6、IL-1β和TNF-α的含量较AngⅡ+miR-21组升高(P<0.05)。结论过表达miR-21能够通过NF-κB信号通路调控AngⅡ诱导的HMC细胞增殖和炎症因子的表达。To investigate the effects of miR-21 on angiotensin Ⅱ(Ang Ⅱ)-induced proliferation and inflammatory factors production of glomerular membrane cells, human glomerular membrane cells(HMC) were cultured in vitro with Ang Ⅱ, and the miR-21 mimics were transferred in the HMC cells induced by Ang II. Realtime fluorescence quantitative PCR was used to detect miR-21 expression level;Thiazolam(MTT) method was employed to detect cell proliferation ability;while Western blot was used to detect the core molecules NF-κBα and p65 proteins and phosphorylation levels of the NF-κB signaling pathway. Enzyme linked immunoadsorption(ELISA)was used to detect the content of inflammatory factors interleukin-6(IL-6), interleukin-1 β(IL-1 β) and tumor necrosis factor-α(TNF-α). The effect of the NF-κB signaling pathway activator PMA on the expression of inflammatory factors was detected. Data showed that compared with control group, HMC cells of Ang Ⅱ group demonstrated lower level of miR-21(P<0.05), increased cell proliferation ability(P<0.05), and higher expression of inflammatory factors IL-6, IL-1 β, and TNF-α(P<0.05). Furthermore, after the over-expression of miR-21 in the Ang II group, the levels of phosphorylation of the IκBα and p65 proteins were significantly reduced(P<0.05), the cell proliferation ability was significantly reduced(P<0.05), and the expression of IL-6, IL-1 β and TNF-α were also reduced(P<0.05). NF-κB signal pathway activator could reverse the reduce of IL-6, IL-1 β and TNF-α in Ang II + miR-21 group(P<0.05). Taken together, overexpression of miR-21 regulates Ang Ⅱ-induced HMC cell proliferation and inflammatory factor expression via NF-κB signaling pathway.

关 键 词：MIR-21 NF-ΚB信号通路 肾小球系膜细胞 炎症反应 糖尿病肾病 

分 类 号：R-332[医药卫生] R593.242