| 王 俭,樊 霞,贾世英,刘 静,孟宪猛,黄海东,陈 悦.miR-21在大鼠心肌细胞缺血再灌注损伤中的作用机制研究[J].,2022,(16):3030-3035 |
| miR-21在大鼠心肌细胞缺血再灌注损伤中的作用机制研究 |
| Study on the Mechanism of miR-21 in Rat Myocardial Ischemia-Reperfusion Injury |
| 投稿时间：2022-03-23 修订日期：2022-04-18 |
| DOI：10.13241/j.cnki.pmb.2022.16.007 |
| 中文关键词: miR-21 大鼠 心肌细胞 缺血再灌注损伤 作用机制 |
| 英文关键词: miR-21 Rat Myocardial cell Ischemia reperfusion injury Mechanism |
| 基金项目:辽宁省自然科学基金项目（2017-ZD-0323） |
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| 中文摘要: |
| 摘要 目的：探讨微小RNA-21（miR-21）在大鼠心肌细胞缺血再灌注损伤中的作用机制。方法：选取50只SPF级Wistar大鼠并随机分为5组（n=10），分别为对照组、模型组、模型+阴性对照组、模型+ miR-21组和模型+ miR-21抑制物组。通过结扎大鼠左冠前降支进行建模。建模成功后采用高频彩色超声诊断仪检查各组大鼠的心脏功能指标：心脏射血分数（EF）、左心室收缩期峰值压力（LVSP）、左室舒张末压（LVEDP）和缩短分数（FS）。检测各组大鼠心肌梗死面积和心肌细胞凋亡率。采用酶联免疫吸附试验（ELISA）测定各组心肌组织中肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）和白细胞介素-10（IL-10）的含量。采用反转录聚合酶链式反应（RT-PCR）检测心肌组织中miR-21的表达水平。蛋白免疫印迹试验（Western Blot）检测各组大鼠心肌凋亡蛋白和TLR4/NF-κB表达水平。结果：模型组大鼠出现心肌梗死，提示建模成功，建模后大鼠心肌组织中miR-21的表达水平显著下降，提示miR-21可能具有保护心肌细胞的作用。建模成功后，EF、LVSP和FS下降，LVEDP升高，心肌细胞凋亡率显著升高，TNF-α和IL-6表达水平显著升高，IL-10显著下降，Bcl-2/Bax表达下降，Caspase-3表达升高，大鼠心肌细胞TLR4和NF-κB蛋白磷酸化表达水平升高，而模型+ miR-21组上述指标均得到改善。结论：大鼠心肌细胞缺血再灌注损伤导致miR-21表达降低，而过表达miR-21能有效抑制TLR4/NF-κB信号通路，降低大鼠心肌凋亡水平和炎症因子的释放，从而发挥保护心肌细胞的作用。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the mechanism of microrna-21 (miR-21) in rat myocardial ischemia-reperfusion injury. Methods: Fifty SPF Wistar rats were selected and randomly divided into 5 groups (n=10), including control group, model group, model + negative control group, model + miR-21 group and model + miR-21 inhibitor group. The model was established by ligation of the left anterior descending coronary artery in rats. After the successful modeling, high-frequency color ultrasound for small animals was used to examine the cardiac function indexes of rats in each group: cardiac ejection fraction (EF), left ventricular peak systolic pressure (LVSP), left ventricular end diastolic pressure (LVEDP) and shortening fraction (FS). Myocardial infarction size and apoptosis rate of myocardial cells were measured. The contents of tumor necrosis factor -α (TNF-α), interleukin-6 (IL-6) and interleukin-10 (IL-10) in myocardial tissues were determined by enzyme-linked immunosorbent assay (ELISA). Reverse transcription polymerase chain reaction (RT-PCR) was used to detect the expression level of miR-21 in myocardial tissue. The expression levels of apoptosis protein and TLR4/NF-κB were detected by western blot. Results: Myocardial infarction occurred in rats in the model group, indicating that the modeling was successful. After modeling, the expression level of miR-21 in rat myocardial tissue decreased significantly, suggesting that miR-21 may protect myocardial cells. After successful modeling, EF, LVSP and FS decreased, LVEDP increased, apoptosis rate of myocardial cells significantly increased, TNF-α and IL-6 expression significantly increased, IL-10 significantly decreased, Bcl-2/Bax expression decreased, caspase-3 expression increased. The expression levels of TLR4 and NF-κB protein phosphorylation increased in rat cardiomyocytes, while the above indicators were improved in model + miR-21 group. Conclusion: Ischemia reperfusion injury of rat cardiomyocytes leads to decreased expression of miR-21, Overexpression of miR-21 can effectively inhibit the TLR4/NF-κB signaling pathway, reduce the level of myocardial apoptosis and the release of inflammatory factors, and thus play a protective role in myocardial cells. |
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