Zhang Liqin, Zhu Huajie, Teng Xianlin, Sheng Xiaosheng, Yu Beiwei
Department of Laboratory, Jinhua People's Hospital, Jinhua, People's Republic of China.
Department of obstetrics and gynecology, 2nd Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, People's Republic of China.
Autoimmunity. 2021 Jun;54(4):195-203. doi: 10.1080/08916934.2021.1910812. Epub 2021 May 27.
Acute myocardial infarction (AMI) is a severe cardiovascular condition. Blocking the apoptosis of myocardial cells may mitigate AMI. Excessive expression of Stanniocalcin-1 (STC1) plays a protective role in the heart by inhibiting myocardial cell apoptosis. Here, we looked at the mechanism by which miR-382-5p regulates STC1 and affects myocardial cell apoptosis after AMI.
An AMI mouse model with a descending anterior ligament coronary artery and an HL-1 cell model with reproducible hypoxia/reoxygenation (H/R) were established. For pathological changes in myocardial tissues, terminal deoxynucleotidyl transferase dUTP nick end labelling staining and haematoxylin and eosin staining were performed. STC1 mRNA and miR-382-5p levels were measured using quantitative real-time PCR. Protein levels of STC1 and apoptosis-related proteins were measured by western blotting. The 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di- phenytetrazoliumromide assay was used to detect cell viability, and a dual-luciferase reporter assay was carried out to verify potential targets of miR-382-5p.
The level of miR-382-5p was raised in myocardial tissues of AMI mice and H/R-induced HL-1 cells. Compared with the control group, the myocardial tissue cells in the AMI group were disordered, with evident necrosis of myocardial cells, apoptosis and inflammatory infiltration. Interference with miR-382-5p inhibited myocardial cell apoptosis after H/R, as well as inferior lactate dehydrogenase. Also, miR-382-5p adversely regulated STC1 and the expression of STC1 was increased after transfection with miR-382-5p antagomir. Furthermore, interference with miR-382-5p reduced myocardial cell apoptosis after H/R by increasing the expression level of STC1.
To summarise, our study showed an increase in miR-382-5p in myocardial tissues in the AMI mouse model. Interference with miR-382-5p reduced apoptosis of myocardial cells after AMI and the effect was achieved by increasing STC1 expression.
急性心肌梗死（AMI）是一种严重的心血管疾病。阻断心肌细胞凋亡可能减轻急性心肌梗死。鲟鱼钙蛋白-1（STC1）的过度表达通过抑制心肌细胞凋亡在心脏中发挥保护作用。在此，我们研究了miR-382-5p调节STC1并影响急性心肌梗死后心肌细胞凋亡的机制。
建立了前降支冠状动脉急性心肌梗死小鼠模型和可重复性缺氧/复氧（H/R）的HL-1细胞模型。对于心肌组织的病理变化，进行了末端脱氧核苷酸转移酶dUTP缺口末端标记染色和苏木精-伊红染色。使用定量实时PCR测量STC1 mRNA和miR-382-5p水平。通过蛋白质印迹法测量STC1和凋亡相关蛋白的水平。采用3-（4,5）-二甲基噻唑-（-z-y1）-3,5-二苯基四氮唑溴盐法检测细胞活力，并进行双荧光素酶报告基因检测以验证miR-382-5p的潜在靶标。
急性心肌梗死小鼠心肌组织和H/R诱导的HL-1细胞中miR-382-5p水平升高。与对照组相比，急性心肌梗死组心肌组织细胞排列紊乱，心肌细胞明显坏死、凋亡及炎性浸润。干扰miR-382-5p可抑制H/R后心肌细胞凋亡以及降低乳酸脱氢酶水平。此外，miR-382-5p对STC1起负向调节作用，转染miR-382-5p拮抗剂后STC1表达增加。此外，干扰miR-382-5p通过增加STC1表达水平减少H/R后心肌细胞凋亡。
总之，我们的研究表明急性心肌梗死小鼠模型心肌组织中miR-382-5p增加。干扰miR-382-5p可减少急性心肌梗死后心肌细胞凋亡，且该作用通过增加STC1表达实现。
