Department of Cardiovascular Medicine, Taizhou People's Hospital, Taizhou, China.
Eur Rev Med Pharmacol Sci. 2019 Oct;23(19):8580-8587. doi: 10.26355/eurrev_201910_19174.
The aim of this study was to investigate the effect of micro ribonucleic acid (miR)-497 on myocardial cell apoptosis in rats with myocardial ischemia/reperfusion (I/R) through the mitogen-activated protein kinase (MAPK)/extracellular regulated protein kinase (ERK) signaling pathway.
A rat model of myocardial I/R was established, myocardial cells were extracted, and miR-497 was inhibited by inhibitors and overexpressed using miRNA mimics. The cell apoptosis rate was detected by flow cytometry and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. The interaction between miR-497 and ERK was determined by dual-luciferase reporter gene assay. The change in the protein level was measured via Western blotting (WB).
Up-regulation of miR-497 promoted myocardial cell apoptosis, and the 3'-untranslated region (3'-UTR) of ERK was highly conserved to combine with miR-497. The luciferase reporter gene assay showed that the transfection of miR-497 could significantly inhibit the relative luciferase activity in cells.
MiR-497 overexpression significantly down-regulated the ERK expression at messenger RNA (mRNA) and protein levels in cells. MiR-497 plays an important role in regulating I/R injury-induced myocardial cell apoptosis by targeting the ERK-induced apoptosis pathway.
本研究旨在通过丝裂原活化蛋白激酶（MAPK）/细胞外调节蛋白激酶（ERK）信号通路，探讨微小 RNA（miR）-497 对心肌缺血再灌注（I/R）大鼠心肌细胞凋亡的影响。
建立大鼠心肌 I/R 模型，提取心肌细胞，采用抑制剂抑制 miR-497 表达，利用 miRNA 模拟物过表达 miR-497。采用流式细胞术和末端脱氧核苷酸转移酶 dUTP 缺口末端标记（TUNEL）法检测细胞凋亡率。采用双荧光素酶报告基因检测 miR-497 与 ERK 之间的相互作用。采用 Western blot（WB）检测蛋白水平的变化。
miR-497 上调促进心肌细胞凋亡，ERK 的 3'-非翻译区（3'-UTR）与 miR-497 高度保守，可结合。荧光素酶报告基因检测显示，miR-497 转染可显著抑制细胞内相对荧光酶活性。
miR-497 过表达可显著下调细胞中 ERK 的信使 RNA（mRNA）和蛋白水平表达。miR-497 通过靶向 ERK 诱导的凋亡通路，在调节 I/R 损伤诱导的心肌细胞凋亡中发挥重要作用。
