Wang Xuejun, Su Jinwen, Lin Zhikang, Liu Kangyong, Zhuang Yu
Department of Cardiology, Shanghai University of Medicine and Health Sciences Affiliated Zhoupu Hospital, Shanghai 201318, P.R. China.
Department of Cardiology, Shanghai Ninth People's Hospital Affiliated to Shanghai JiaoTong University School of Medicine, Shanghai 200011, P.R. China.
Exp Ther Med. 2022 Jan;23(1):62. doi: 10.3892/etm.2021.10984. Epub 2021 Nov 22.
Myocardial infarction (MI), the leading cause of death among patients with cardiovascular diseases, is characterized by acute cardiac muscle injury due to severe impairment of the coronary blood supply, which may lead to cardiogenic shock and cardiac arrest. Particularly interesting new cysteine histidine rich 1 (PINCH1) protein, a key component of the integrin signaling pathway, interacts with several proteins and serves a vital role in numerous cellular processes, including cytoskeleton remodeling, cell proliferation and cell migration. To investigate the role of PINCH1 in heart injury in the present study, PINCH1 was knocked out in the myocardial tissue of mice (age, 18 weeks) to induce MI. In addition, cell viability, migration and apoptosis, as well as the expression levels of NF-κB-associated proteins were determined in murine HL1 cardiomyocytes with a conditional PINCH1 shRNA using Cell Counting Kit-8, Transwell, flow cytometry and western blot assays, respectively. Furthermore, the cardiac expansion and myocardial fibrosis in PINCH1 knockout mice was investigated by performing morphological and histological examinations. Additionally, the murine ventricular myocardial ultrastructure was evaluated using an electron microscope, and the cardiomyocyte apoptotic rate and expression levels of NF-κB-related proteins were determined using TUNEL and western blot assays, respectively. The results showed that the apoptotic rate in the PINCH1 knockdown group was significantly increased. In addition, the protein expression levels of NF-κB signaling pathway-related proteins, including NF-κB, myeloid differentiation factor 88, TNF-α and caspase-3, were significantly increased in the PINCH1 knockdown group compared with the wild-type group, but the protein expression of MMP2 and MMP9 were the opposite. Overall, the and results revealed that PINCH1 knockout in mice significantly aggravated MI via the NF-κB signaling pathway.
心肌梗死（MI）是心血管疾病患者死亡的主要原因，其特征是由于冠状动脉血液供应严重受损导致急性心肌损伤，这可能会导致心源性休克和心脏骤停。特别值得关注的是富含半胱氨酸组氨酸的新蛋白1（PINCH1），它是整合素信号通路的关键组成部分，与多种蛋白质相互作用，并在众多细胞过程中发挥重要作用，包括细胞骨架重塑、细胞增殖和细胞迁移。为了在本研究中探究PINCH1在心脏损伤中的作用，在小鼠（18周龄）的心肌组织中敲除PINCH1以诱导心肌梗死。此外，分别使用细胞计数试剂盒-8、Transwell、流式细胞术和蛋白质印迹分析，测定了具有条件性PINCH1短发夹RNA的小鼠HL1心肌细胞的细胞活力、迁移和凋亡以及NF-κB相关蛋白的表达水平。此外，通过进行形态学和组织学检查，研究了PINCH1基因敲除小鼠的心脏扩张和心肌纤维化。另外，使用电子显微镜评估小鼠心室心肌超微结构，并分别使用TUNEL和蛋白质印迹分析测定心肌细胞凋亡率和NF-κB相关蛋白的表达水平。结果显示，PINCH1敲低组的凋亡率显著增加。此外，与野生型组相比，PINCH1敲低组中NF-κB信号通路相关蛋白的表达水平显著增加，包括NF-κB、髓样分化因子88、TNF-α和caspase-3，但MMP2和MMP9的蛋白表达情况则相反。总体而言，上述结果表明，小鼠中PINCH1基因敲除通过NF-κB信号通路显著加重了心肌梗死。
