Department of Gerontology, Shibei Hospital of Jing'an District, Shanghai 200443, P.R. China.
Department of Gerontology, Shanghai Ninth People's Hospital, Shanghai JiaoTong University School of Medicine, Shanghai 200011, P.R. China.
Mol Med Rep. 2021 Feb;23(2). doi: 10.3892/mmr.2020.11790. Epub 2020 Dec 20.
Angiotensin II (AngII) is a central signaling molecule of the renin‑angiotensin system that serves a vital role in myocardial fibrosis (MF). The present study aimed to investigate the effects of matrix metalloproteinase (MMP)3 on MF progression. To induce cellular fibrosis, H9C2 rat myocardial cells were treated with AngII for 24 h. Subsequently, cells were treated with levocarnitine, or transfected with small interfering (si)RNA‑negative control or siRNA‑MMP3 (1/2/3). Cell viability, apoptosis and migration were assessed by performing Cell Counting Kit‑8, flow cytometry and Transwell assays, respectively. Reverse transcription‑quantitative PCR (RT‑qPCR) and western blotting were performed to determine the expression levels of MF biomarkers, including disease‑, apoptosis‑ and oxidative stress‑related genes. Compared with the control group, AngII significantly inhibited H9C2 cell viability and migration, and significantly increased H9C2 cell apoptosis (P<0.05). However, compared with AngII‑treated H9C2 cells, MMP3 knockdown significantly inhibited fibrotic H9C2 cell viability and migration, but increased fibrotic H9C2 cell apoptosis (P<0.05). The RT‑qPCR results demonstrated that MMP3 knockdown significantly downregulated the expression levels of AXL receptor tyrosine kinase, AngII receptor type 1, α‑smooth muscle actin and Collagen I in AngII‑treated H9C2 cells (P<0.05). Moreover, compared with AngII‑treated cells, MMP3 knockdown significantly decreased Bcl‑2 expression levels , but significantly increased caspase‑3 and p53 expression levels in AngII‑treated cells (P<0.05). Additionally, compared with AngII‑treated cells, MMP3 knockdown significantly decreased MMP3, MMP9, STAT3, p22Phox and p47Phox expression levels in AngII‑treated cells (P<0.05). The present study indicated that MMP3 knockdown altered myocardial fibroblast cell viability, migration and apoptosis by regulating apoptosis‑ and oxidative stress‑related genes, thus delaying MF progression.
血管紧张素 II（AngII）是肾素-血管紧张素系统的核心信号分子，在心肌纤维化（MF）中发挥着重要作用。本研究旨在探讨基质金属蛋白酶 3（MMP3）对 MF 进展的影响。为了诱导细胞纤维化，将 H9C2 大鼠心肌细胞用 AngII 处理 24 小时。随后，用左卡尼汀或转染小干扰 RNA-阴性对照或 siRNA-MMP3（1/2/3）处理细胞。通过细胞计数试剂盒-8、流式细胞术和 Transwell 测定分别评估细胞活力、凋亡和迁移。采用逆转录-定量 PCR（RT-qPCR）和蛋白质印迹法检测 MF 生物标志物，包括疾病、凋亡和氧化应激相关基因的表达水平。与对照组相比，AngII 显著抑制 H9C2 细胞活力和迁移，显著增加 H9C2 细胞凋亡（P<0.05）。然而，与 AngII 处理的 H9C2 细胞相比，MMP3 敲低显著抑制纤维性 H9C2 细胞活力和迁移，但增加纤维性 H9C2 细胞凋亡（P<0.05）。RT-qPCR 结果表明，MMP3 敲低显著下调 AngII 处理的 H9C2 细胞中 AXL 受体酪氨酸激酶、血管紧张素 II 受体 1、α-平滑肌肌动蛋白和 Collagen I 的表达水平（P<0.05）。此外，与 AngII 处理的细胞相比，MMP3 敲低显著降低 AngII 处理的细胞中 Bcl-2 的表达水平，但显著增加 caspase-3 和 p53 的表达水平（P<0.05）。此外，与 AngII 处理的细胞相比，MMP3 敲低显著降低 AngII 处理的细胞中 MMP3、MMP9、STAT3、p22Phox 和 p47Phox 的表达水平（P<0.05）。本研究表明，MMP3 敲低通过调节凋亡和氧化应激相关基因改变心肌成纤维细胞的活力、迁移和凋亡，从而延缓 MF 进展。
