Dong Hua, Dong Shimin, Zhang Lisi, Gao Xueping, Lv Guangwei, Chen Wei, Shao Suxia
Department of Histology and Embryology, Hebei Medical University, Shijiazhuang, 050017, China.
Department of Emergency, the 3rd Hospital of Hebei Medical University, Shijiazhuang, 050051, China.
Anat Rec (Hoboken). 2016 Oct;299(10):1348-57. doi: 10.1002/ar.23396. Epub 2016 Jul 18.
The miRNAs play important roles in regulating myocardial fibrosis. The purpose of this study was to determine the potential roles of microRNA-214 (miR-214) in cardiac fibrosis in vitro and in vivo. In vitro experiment, Ang II-induced cardiac fibroblasts (CFBs) are transfected with pre-miR-214, anti-miR-214 and their oligo controls. Gene expression was checked by Quantitative realtime-PCR (qRT-PCR) and western blotting. In the present experiment, compared with controls, expressions of collagen type I (COL I), collagen type III (COL III), transforming growth factor (TGF)-β1, and tissue inhibitors of metalloproteinase (TIMP)-1 were all increased, but matrix metalloproteinase (MMP)-1 was reduced in CFB by Ang II treatment at both mRNA and protein levels, and these alterations were found reversed by miR-214 transfection. In vivo, an anterior transmural acute myocardial infarction (AMI) was created by occlusion of the left anterior descending coronary artery after Ad-pre-miR-214, Ad-anti-miR-214 or Ad-GFP was delivered separately. Four weeks after AMI, protein contents of COL I, COL III and TGF-β1 in tissue from border area were found increased after AMI, but impaired by overexpression of miR-214. While the expression of MMP-1 was increased by miR-214 stimulation but decreased by miR-214 inhibition. These results suggested that miR-214 exerts cardio-protective effects by inhibition of fibrosis and the inhibitory effect involves TGF-β1 suppression and MMP-1/TIMP-1 regulation. Anat Rec, 299:1348-1357, 2016. © 2016 Wiley Periodicals, Inc.
微小RNA（miRNA）在调节心肌纤维化中发挥着重要作用。本研究的目的是确定微小RNA-214（miR-214）在体外和体内心脏纤维化中的潜在作用。在体外实验中，用前体miR-214、抗miR-214及其寡核苷酸对照转染血管紧张素II诱导的心脏成纤维细胞（CFB）。通过定量实时聚合酶链反应（qRT-PCR）和蛋白质印迹法检测基因表达。在本实验中，与对照组相比，血管紧张素II处理使CFB中I型胶原（COL I）、III型胶原（COL III）、转化生长因子（TGF）-β1和金属蛋白酶组织抑制剂（TIMP）-1的表达在mRNA和蛋白质水平均增加，但基质金属蛋白酶（MMP）-1减少，而miR-214转染可逆转这些改变。在体内，分别递送腺病毒前体miR-214、腺病毒抗miR-214或腺病毒绿色荧光蛋白（Ad-GFP）后，通过结扎左前降支冠状动脉造成前壁透壁性急性心肌梗死（AMI）。AMI后四周，发现梗死边缘区组织中COL I、COL III和TGF-β1的蛋白含量在AMI后增加，但miR-214过表达可使其受损。而MMP-1的表达在miR-214刺激下增加，但在miR-214抑制下减少。这些结果表明，miR-214通过抑制纤维化发挥心脏保护作用，且这种抑制作用涉及TGF-β1的抑制和MMP-1/TIMP-1的调节。《解剖学记录》，299:1348 - 1357，2016年。©2016威利期刊公司。
