miR-146a对肺纤维化小鼠成纤维细胞增殖的影响及其机制  

Effect of microRNA-146a on proliferation of pulmonary fibroblasts of mice

作　　者：石宇红[1] 任亚飞[1] 蒋亚男 许佳[1] 李宝贞[1] 朱芳晓[1] 莫汉有[1] SHI Yuhong;REN Yafei;JIANG Yanan;XU Jia;LI Baozhen;ZHU Fangxiao;MO Hanyou(The Affiliated Hospital of Guilin Medical University,Guilin 541002,China)

机构地区：[1]桂林医学院附属医院,广西桂林541002

出　　处：《山东医药》2018年第36期13-16,共4页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(81460257)

摘　　要：目的探讨微小RNA-146a(miR-146a)对肺纤维化小鼠成纤维细胞增殖的影响及其机制。方法选择C57BL/6小鼠30只,随机分为模型组和假手术组各15只。模型组经气管滴入博来霉素建立肺纤维化模型,假手术组经气管滴入等体积生理盐水。造模第28天,采用RT-PCR法检测两组肺组织miR-146a相对表达量。提取并培养造模第28天模型组肺成纤维细胞,随机分为miR-146a过表达组、对照组、空白组,分别给予miR-146a mimics(miR-146a激活剂)、miR-146a阴性对照物及缓冲液转染。转染24 h,再培养36 h,收集细胞,采用CCK-8法检测细胞增殖率,采用Western blotting法检测转化生长因子β1(TGF-β1)、核因子κB(NF-κB) p65及NF-κB抑制蛋白α(IκBα)蛋白表达。结果模型组肺组织miR-146a相对表达量明显低于假手术组(P <0. 01)。miR-146a过表达组细胞增殖率为(13. 47±1. 83)%,对照组为(33. 87±1. 32)%,空白组为(32. 57±1. 54)%,miR-146a过表达组低于其他两组(P均<0. 05),而对照组与空白组比较P> 0. 05。与对照组和空白组比较,miR-146a过表达组TGF-β1、IκBα蛋白相对表达量均降低(P <0. 05),NF-κB p65蛋白相对表达量升高(P均<0. 05),而对照组与空白组比较P> 0. 05。结论 miR-146a可抑制肺纤维化小鼠成纤维细胞增殖,其机制可能与阻止细胞内NF-κB核转位有关。Objective To investigate the effect of microRNA(miR)-146a on proliferation of fibroblasts in mice with pulmonary fibrosis.Methods C57BL/6 mice of SPF grade(n=30)were randomly divided into the model group and the sham operation group,with 15 in each.The model group was treated with bleomycin through intratracheal instillation,and the control group was treated with normal saline.The relative expression of miR-146a in the lung tissues was detected by re-al-time PCR on the 28th day of modeling.Lung fibroblasts in the model group were extracted and cultured on the 28th day after the model establishment,and were randomly divided into the miR-146a overexpression group,control group and blank group,which were transfected with miR-146a mimics(miR-146a activator),miR-146a negative control and buffer solution for 48 h.The cell proliferation rate was detected by CCK-8,the cell viability was assessed by CCK-8.Expression of trans-forming growth factor-p1(TGF-p1),NF-kB p65 protein and inhibitory NF-kB protein a(iKBa)in the pulmonary fibro-blasts were detected by Western blotting.Results The relative expression of miR-146a in the lung tissues of the model group was significantly lower than that in the sham operation group(P<0.01).The cell proliferation rates of the miR-146a overexpression group,control group,and the blank group were 13.47%±1.83%,33.87%±1.32%,and 32.57%±1.54%,respectively.The miR-146a overexpression group was lower than the other two groups(both P<0.05).Com-pared with the control group and the blank group,the relative protein expression of iKBa and TGF-p1 significantly de-creased in the miR-146a overexpression group,while the expression of NF-kB-p65 significantly increased at the same time(P<0.05).Conclusion The miR-146a inhibits the proliferation of fibroblasts in mice with pulmonary fibrosis by inhibi-ting NF-kB nuclear translocation.

关 键 词：肺纤维化 肺成纤维细胞 微小RNA-146a 转化生长因子β1 核因子ΚBP65 核因子κB抑制蛋白α 小鼠 

分 类 号：R563[医药卫生—呼吸系统]