微小RNA-199a-5p通过靶向SIRT1促进心肌纤维化相关基因表达

发布时间：2018-06-20 14:10

  本文选题：心肌纤维化 + 成纤维细胞　； 参考：《中国病理生理杂志》2017年10期

【摘要】：目的:研究微小RNA-199a-5p(miR-199a-5p)对心肌成纤维细胞中纤维化相关基因表达的调控作用及其可能作用的靶基因。方法:原代分离并体外培养成体C57BL/6小鼠心肌成纤维细胞;双萤光素酶报告基因实验检测miR-199a-5p与潜在靶基因沉默信息调节因子1(SIRT1)3’端非翻译区(3’-UTR)的结合作用;实时荧光定量PCR(RT-q PCR)和Western blot法分别检测SIRT1以及纤维化标志物胶原蛋白(Col)1a1、Col3a1和α-平滑肌肌动蛋白(α-SMA)的mRNA和蛋白表达。结果:在血管紧张素Ⅱ(AngⅡ)诱导的小鼠心肌成纤维细胞中,Col1a1、Col3a1和α-SMA的表达增强,miR-199a-5p表达上调。在心肌成纤维细胞中过表达miR-199a-5p可以增强Col1a1、Col3a1和α-SMA的表达。双萤光素酶报告基因实验显示miR-199a-5p与SIRT1 3’-UTR有结合作用。RT-q PCR和Western blot结果证实miR-199a-5p可在转录水平抑制SIRT1表达。过表达miR-199a-5p和沉默SIRT1均能一致性促进心肌成纤维细胞中Col1a1、Col3a1和α-SMA的表达。抑制AngⅡ诱导的小鼠心肌成纤维细胞中NF-κB激活,可显著降低miR-199a-5p表达。结论:SIRT1是miR-199a-5p的作用靶基因,并介导miR-199a-5p促进纤维化标志物Col1a1、Col3a1和α-SMA的表达。
[Abstract]:Aim: to study the regulatory effect of microRNA-199a-5pnmiR-199a-5p on the expression of fibrogenic genes in cardiac fibroblasts and the possible target genes. Methods: primary cultured adult C57BL / 6 mouse myocardial fibroblasts were isolated and cultured in vitro, and the binding of miR-199a-5p to potential target gene silencing information regulator 1SIRT1 3'untranslated region (3'UTRs) was detected by double luciferase reporter gene experiment. The mRNA and protein expressions of SIRT1, collagen marker Col1, Col3a1 and 伪 -smooth muscle actin (伪 -SMAs) were detected by real-time fluorescence quantitative PCR and Western blot, respectively. Results: the expression of Col1a1, Col3a1 and 伪 -SMA in myocardial fibroblasts induced by angiotensin 鈪，

本文编号：2044529