MicroRNA-101对肺尘埃沉着病肺纤维化影响的研究

Study of effect of microRNA-101 on pulmonary fibrosis in pneumoconiosis

目的 研究microRNA-101(miR-101)对肺尘埃沉着病肺纤维化的作用机制,为肺尘埃沉着病的二级预防提供新的策略和方法。方法 通过体外实验初步探讨miR-101在SiO_(2)粉尘诱导肺纤维化中的调节作用。选取SPF级健康成年雄性小鼠18只,将其分为空白对照组、生理盐水组和SiO_(2)组,每组6只。采用SiO_(2)粉尘混悬液构建小鼠肺尘埃沉着病模型,HE染色观察大鼠肺组织形态学改变;采用实时定量PCR、蛋白质印迹法检测各组小鼠肺组织中miR-101和Ⅰ型胶原蛋白(ColⅠ)表达水平。结果 模型组在28 d形成肺尘埃沉着病模型。空白对照组及生理盐水组小鼠肺组织结构完整,肺泡间隔均匀,无间质纤维组织增生,SiO_(2)组小鼠肺部组织发生纤维化,肺部结构被破坏,肺间质和肺泡内有大量的细胞浸润,并伴有炎症反应,血管壁增厚。SiO_(2)组的miR-101表达水平低于空白对照组,而ColⅠ蛋白水平高于空白对照组,差异均有统计学意义(P<0.05)。结论 miR-101通过参与抑制炎症因子的释放,从而实现对肺尘埃沉着病模型细胞的纤维化的抑制。

Objective To investigate the effect of microRNA-101(miR-101)on delaying pulmonary fibrosis of pneumoconiosis and its mechanism and to provide new strategies and methods for the secondary prevention of pneumoconiosis.Methods The regulatory role of miR-101 in SiO_(2) dust-induced pulmonary fibrosis was preliminarily explored by in vitro experiments.Eighteen healthy adult male mice of SPF grade were selected and divided into blank control group,saline group and SiO_(2) group,6 mice in each group.The mouse pneumoconiosis model was constructed by using SiO_(2) dust suspension,and the morphological changes of mouse lung tissue were observed by HE staining,the expression levels of miR-101 and Col I were detected by real-time quantitative PCR and Western blotting in the lung tissues of each group of mice.Results The model group formed a pneumoconiosis model at 28 d.In the blank control group and the saline control group,the lung tissue structure was intact,the alveolar intervals were uniform,and there was no interstitial fibrous tissue proliferation,while in the SiO_(2) group,fibrosis occurred in the lung tissue,the lung structure was damaged,and there was a large number of cells infiltrating in the interstitium and alveoli,accompanied by inflammatory reaction and thickening of the vascular wall.miR-101 expression level was lower than that of the blank control group in the SiO_(2) group,and the ColⅠprotein level was higher than that of the blank control group,and the differences were all statistically significant(P<0.05).Conclusion MiR-101 can inhibit the fibrosis of pneumoconiosis model cells by inhibiting the release of inflammatory factors.