Group of Molecular and Cellular Cardiology, Department of Circulation and Medical Imaging, Faculty of Medicine and Health, Norwegian University of Science and Technology, Trondheim, Norway.
St. Olav's University Hospital, Trondheim, Norway.
Am J Physiol Cell Physiol. 2020 Jan 1;318(1):C94-C102. doi: 10.1152/ajpcell.00204.2019. Epub 2019 Oct 16.
Matrix metalloproteinases (MMP) are important for cardiac remodeling. Recently, microRNA (miR)-451a has been found to inhibit the expression of both MMP-2 and MMP-9 in human malignancies, but its role in cardiomyocytes has not been explored. We hypothesized that miR-451a modulates MMP-2 and MMP-9 levels in human cardiomyocytes. The role of miR-451a on regulation of MMP-2 and MMP-9 was evaluated in two separate pathological models using Cor.4U human inducible pluripotent stem cell-derived cardiomyocytes (hiPS-CMs): ) endothelin-1 (ET-1), and ) 48-h hypoxia (1% O). Both models were transfected with synthetic miR-451a mimics or scramble control. Expression of both mRNA and miR was determined by quantitative real-time polymerase chain reaction and protein activity by (MMP-2/9) activity assay. Bioinformatic analyses were performed using Targetscan 7.1 and STRING 10.5. hiPS-CMs stimulated by hypoxia increased both MMP-2 and MMP-9 expression levels compared with normoxia ( < 0.05), whereas ET-1 stimulation only increased the MMP-9 level compared with vehicle controls ( < 0.05). miR-451a mimics prevented the increase of MMP-2 and MMP-9 expression in both models. Protein activity of MMP-2 and MMP-9 was confirmed to be lower following treatment with miR-451a mimic compared with scramble-controls. Six of 28 predicted gene transcripts of miR-451a were linked to MMP-2 and MMP-9; Macrophage migration inhibitory factor (MIF) was the only predicted target of miR-451a that was increased by ET-1 and hypoxia and reduced following miR-451a mimic transfection. miR-451a prevent the increase of MMP-2 and MMP-9 in human cardiomyocytes during pathological stress. The modulation by miR-451a on MMP-2 and MMP-9 is caused by MIF.
基质金属蛋白酶（MMP）对于心脏重构非常重要。最近，研究发现微小 RNA（miR）-451a 可抑制人类恶性肿瘤中 MMP-2 和 MMP-9 的表达，但它在心肌细胞中的作用尚未得到探索。我们假设 miR-451a 可调节人心肌细胞中 MMP-2 和 MMP-9 的水平。我们使用 Cor.4U 人诱导多能干细胞衍生的心肌细胞（hiPS-CM）的两个独立病理模型评估了 miR-451a 对 MMP-2 和 MMP-9 调节的作用：）内皮素-1（ET-1），和）48 小时缺氧（1% O）。这两种模型都用合成的 miR-451a 模拟物或乱序对照物进行了转染。通过定量实时聚合酶链反应确定 mRNA 和 miR 的表达，通过（MMP-2/9）活性测定法确定蛋白活性。使用 Targetscan 7.1 和 STRING 10.5 进行生物信息学分析。与正常氧相比，缺氧刺激的 hiPS-CM 增加了 MMP-2 和 MMP-9 的表达水平（<0.05），而 ET-1 刺激仅增加了与载体对照相比 MMP-9 的水平（<0.05）。miR-451a 模拟物可防止两种模型中 MMP-2 和 MMP-9 表达的增加。与 scramble 对照相比，miR-451a 模拟物处理后 MMP-2 和 MMP-9 的蛋白活性证实较低。miR-451a 的 28 个预测基因转录物中有 6 个与 MMP-2 和 MMP-9 相关；巨噬细胞移动抑制因子（MIF）是唯一被 ET-1 和缺氧上调、miR-451a 模拟物转染后下调的 miR-451a 预测靶点。miR-451a 可防止人心肌细胞在病理应激期间 MMP-2 和 MMP-9 的增加。miR-451a 对 MMP-2 和 MMP-9 的调节是由 MIF 引起的。
