miR-21对ADAMTS-1的调节在肺纤维化体内外实验中的研究  

Study of regulation of miR-21 on ADAMTS-1 expression

作　　者：赵红[1] 姚平波[2] 戴利[1] 张平[1] 何军[2] 

机构地区：[1]南华大学护理学院,湖南省衡阳市421001 [2]南华大学附属南华医院,湖南省衡阳市421002

出　　处：《实用医学杂志》2015年第15期2426-2429,共4页The Journal of Practical Medicine

基　　金：湖南省自然科学基金资助项目(编号:14JJ7044);湖南省教育厅资助项目(编号:14B157)

摘　　要：目的:探讨mi R-21在肺纤维化模型中对ADAMTS-1表达水平的调节作用。方法:(1)建立博来霉素诱导的小鼠肺纤维化模型和构建体外肺纤维化模型,应用q RT-PCR检测mi R-21表达水平,Western blot法观察ADAMTS-1蛋白的表达。(2)通过体外转染mi R-21相似剂(mimics)及抑制剂(inhibitors)至NIH3T3细胞,Westem blot法检测ADAMTS-1蛋白的表达。结果:(1)在小鼠肺纤维化模型中,与空白对照组比较,博来霉素处理后的3个时间点,小鼠肺组织mi R-21表达明显上调,而ADAMTS-1蛋白表达明显下调(P<0.01)。(2)在构建体外肺纤维化模型中,NIH3T3细胞经5μg/L的TGF-β1刺激后,ADAMTS-1蛋白表达下调,mi R-21表达量明显上调(P<0.01)。(3)对NIH3T3细胞转染mi R-21相似物及抑制剂后实验结果显示,高表达mi R-21后,NIH3T3细胞ADAMTS-1蛋白表达下调,反之亦然。结论:(1)体内外实验证实,肺纤维过程中mi R-21表达上升,ADAMTS-1蛋白表达下降。(2)mi R-21可负调控细胞ADAMTS-1的表达水平。Objective To investigate the regulatory effect of miR-21 on ADAMTS-1 expression in pulmonary fibrosis model. Methods A mouse model of pulmonary fibrosis was established using bleomycin （BLM） anda model of pulmonary fibrosis was constructed in vitro, the expression level of miR-21 was measured by quantitative real-time polymerase chain reaction （qRT-PCR）, while the protein expression of ADAMTS-1 was measured by Western blot. NIH3T3 were transfected with miR-21 mimics and inhibitor in vitro and the cellular expression of ADAMTS-1 was measured by Western blot. Results Compared with the blank group, in mouse models of pulmonary fibrosis, the miR-21 expressions in lung tissues at three time points after BLM-treatment were significantly up-regulated while an evident decrease in ADAMTS-1 expressions were observed （P 〈 0.01 ）. In vitro pulmonary fibrosis model, NIH3T3 cells after TGF-（31 in concentration 5 μg/L stimulation down- regulated ADAMTS-1 expression and up-regulated miR-21 expression（P 〈 0.01）. NIH3T3 transfected with miR- 21 mimics and inhibitor, up-regulated miR-21 expression, while down-regulated ADAMTS-1 protein expression. Conclusions Up-regulation of miR-21 and Down-regulation of ADAMTS-1 might be involved in the progression of pulmonary fibrosis model; miR-21 could negatively regulate ADAMTS-1 expression.

关 键 词：微小RNA 体内外实验 ADAMTS-1 肺纤维化 

分 类 号：R563[医药卫生—呼吸系统]