Xiang Shaohua, Li Jing, Zhang Zhengfu
Department of Cardiothoracic Surgery, Dianjiang County Hospital of Traditional Chinese Medicine, Chongqing 408300, P.R. China.
Department of Cardiothoracic Surgery, People's Hospital of Changshou, Chongqing 401220, P.R. China.
Exp Ther Med. 2020 Mar;19(3):2067-2074. doi: 10.3892/etm.2020.8455. Epub 2020 Jan 15.
A critical event in cardiac fibrosis is the transformation of cardiac fibroblasts (CFs) into myofibroblasts. MicroRNAs (miRNAs) have been reported to be critical regulators in the development of cardiac fibrosis. However, the underlying molecular mechanisms of action of miRNA (miR)-26b in cardiac fibrosis have not yet been extensively studied. In the present study, the expression levels of miR-26b were downregulated in isoproterenol (ISO)-treated cardiac tissues and CFs. Moreover, miR-26b overexpression inhibited the cell viability of ISO-treated CFs and decreased the protein levels of collagen I and α-smooth muscle actin (α-SMA). Furthermore, bioinformatics analysis and dual luciferase reporter assays indicated that Kelch-like ECH-associated protein 1 (Keap1) was the target of miR-26b, and that its expression levels were decreased in miR-26b-treated cells. In addition, Keap1 overexpression reversed the inhibitory effects of miR-26b on ISO-induced cardiac fibrosis, as demonstrated by cell viability, and the upregulation of collagen I and α-SMA expression levels. Furthermore, inhibition of Keap1 expression led to the activation of nuclear factor erythroid 2-related factor 2 (Nrf2), which induced the transcriptional activation of antioxidant/detoxifying proteins in order to protect against cardiac fibrosis. Taken together, the data demonstrated that miR-26b attenuated ISO-induced cardiac fibrosis via the Keap-mediated activation of Nrf2.
心脏纤维化中的一个关键事件是心脏成纤维细胞(CFs)向肌成纤维细胞的转变。据报道,微小RNA(miRNAs)是心脏纤维化发展过程中的关键调节因子。然而,miRNA(miR)-26b在心脏纤维化中潜在的分子作用机制尚未得到广泛研究。在本研究中,miR-26b的表达水平在异丙肾上腺素(ISO)处理的心脏组织和CFs中下调。此外,miR-26b过表达抑制了ISO处理的CFs的细胞活力,并降低了I型胶原蛋白和α-平滑肌肌动蛋白(α-SMA)的蛋白质水平。此外,生物信息学分析和双荧光素酶报告基因检测表明, Kelch样ECH相关蛋白1(Keap1)是miR-26b的靶标,并且其在miR-26b处理的细胞中的表达水平降低。另外,Keap1过表达逆转了miR-26b对ISO诱导的心脏纤维化的抑制作用,这通过细胞活力以及I型胶原蛋白和α-SMA表达水平的上调得到证明。此外,抑制Keap1表达导致核因子红细胞2相关因子2(Nrf2)的激活,其诱导抗氧化/解毒蛋白的转录激活以预防心脏纤维化。综上所述,数据表明miR-26b通过Keap介导的Nrf2激活减轻了ISO诱导的心脏纤维化。