长链非编码RNA Dnm3os在心肌成纤维细胞活化中的作用研究

The potential role of long non-coding RNA Dnm3os in the activation of cardiac fibroblasts

长链非编码RNA(lncRNA)Dnm3os在肌腱周围组织纤维化和肺纤维化中发挥了重要作用,但其是否参与心肌纤维化过程,目前尚不清楚。为此,本研究利用课题组前期通过胸主动脉缩窄术(TAC)所致的小鼠纤维化模型的心脏组织,以及转化生长因子-β1(TGF-β1)所诱导的体外心肌成纤维细胞活化模型,采用定量聚合酶链式反应(RT-qPCR)、蛋白质印迹法(Western blot)及胶原凝胶收缩等方法,对心肌成纤维细胞活化表型和Dnm3os的表达变化进行鉴定和检测。并通过RNA干扰技术来沉默Dnm3os,以探究其在心肌成纤维细胞活化过程中的作用。结果表明,Dnm3os在小鼠纤维化心肌组织和体外培养活化的心肌成纤维细胞中均表达升高,而沉默其表达则可明显缓解心肌成纤维细胞活化。此外,TGF-β1/Smad2/3通路在心肌成纤维细胞活化过程中被激活;而沉默Dnm3os后,该通路受到抑制。本研究的结果提示,沉默Dnm3os可通过抑制TGF-β1/Smad2/3信号通路来影响心肌成纤维细胞的活化过程。因此,干预lncRNA Dnm3os的表达可能成为治疗心肌纤维化的潜在靶点。

Long non-coding RNA(lncRNA)Dnm3 os plays a critical role in peritendinous fibrosis and pulmonary fibrosis,but its role in the process of cardiac fibrosis is still unclear.Therefore,we carried out study by using the myocardial fibrotic tissues obtained by thoracic aortic constriction(TAC)in an early study of our group,and the in vitro cardiac fibroblast activation model induced by transforming growth factor-β1(TGF-β1).Quantitative real-time polymerase chain reaction(RT-qPCR),Western blot,and collagen gel contraction test were used to identify the changes of activation phenotype and the expression of Dnm3 os in cardiac fibroblasts.Small interfering RNA was used to silence Dnm3 os to explore its role in the activation of cardiac fibroblasts.The results showed that the expression of Dnm3 os was increased significantly in myocardial fibrotic tissues and in the activated cardiac fibroblasts.And the activation of cardiac fibroblasts could be alleviated by Dnm3 os silencing.Furthermore,the TGF-β1/Smad2/3 pathway was activated during the process of cardiac fibroblasts activation,while was inhibited after silencing Dnm3 os.The results suggest that Dnm3 os silencing may affect the process of cardiac fibroblast activation by inhibiting TGF-β1/Smad2/3 signal pathway.Therefore,interfering with the expression of lncRNA Dnm3 os may be a potential target for the treatment of cardiac fibrosis.