| 邢 智,高 颖,帕丽达·阿布来提,李 辉,沙吉旦·阿不都热衣木.miR-21对心肌缺血大鼠心肌细胞TLR-4/NF-κB通路的影响[J].,2023,(5):812-817 |
| miR-21对心肌缺血大鼠心肌细胞TLR-4/NF-κB通路的影响 |
| Effects of miR-21 on TLR-4/NF-κB Pathway in Myocardial Cells of Rats with Myocardial Ischemia |
| 投稿时间：2022-06-21 修订日期：2022-07-18 |
| DOI：10.13241/j.cnki.pmb.2023.05.003 |
| 中文关键词: 微小核糖核酸-21 Toll样受体4 核因子-κB 心肌缺血 凋亡 |
| 英文关键词: miR-21 TLR4 NF-κB Myocardial ischemia Apoptosis |
| 基金项目:新疆维吾尔自治区自然科学基金面上项目（2021D01C310）；新疆医科大学第一附属医院优秀青年人才培养专项（新医大一附院院字【2020】63号） |
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| 中文摘要: |
| 摘要 目的：探究微小核糖核酸-21（miR-21）对心肌缺血大鼠心肌细胞Toll样受体4（TLR4）/核因子-κB（NF-κB）通路的影响。方法：取40只SD大鼠随机分为假手术组（10只）和建模组（30只），建模组大鼠通过左冠状动脉前降支（LAD）结扎术建立心肌缺血大鼠模型，假手术组大鼠仅开胸后不做其他处理即缝合。将建模成功大鼠随机分为对照组（转染miR-NC慢病毒）、沉默组（转染miR-21 antagomir慢病毒）、过表达组（转染miR-21 mimics慢病毒），假手术组注射等量生理盐水。苏木素-伊红（HE）染色观察心肌组织损伤情况；原位末端标记法（TUNEL）检测细胞凋亡率；实时定量聚合酶链反应（RT-qPCR）和蛋白质免疫印迹法（WB）检测TLR-4、NF-κB、半胱氨酸天冬氨酸蛋白酶3（Caspase3）、B淋巴细胞瘤-2基因（Bcl-2）、Bcl-2关联X蛋白（Bax）信使核糖核酸（mRNA）和蛋白表达及磷酸化NF-κB p65（p-NF-κB p65）水平。结果：对照组大鼠心肌纤维排列紊乱，大量心肌细胞肿胀坏死并伴随炎症细胞浸润，沉默组大鼠心肌组织损伤情况进一步加重，而过表达组大鼠心肌组织损伤现象得到明显改善。与假手术组比，对照组、沉默组、过表达组大鼠心肌细胞凋亡率，TLR-4、NF-κB、Caspase-3、Bax mRNA与其蛋白表达，以及p-NF-κB p65水平升高，Bcl-2 mRNA及其蛋白表达降低（P＜0.05）；与对照组比，沉默组大鼠心肌细胞凋亡率，TLR-4、NF-κB、Caspase-3、Bax mRNA表达与其蛋白表达，以及p-NF-κB p65水平升高，Bcl-2 mRNA及其蛋白表达降低（P＜0.05），过表达组大鼠心肌细胞凋亡率，TLR-4、NF-κB、Caspase-3、Bax mRNA与其蛋白表达，以及p-NF-κB p65水平降低，Bcl-2 mRNA及其蛋白表达升高（P＜0.05）。结论：下调miR-21表达可促进TLR-4/NF-κB通路表达，加重心肌缺血大鼠心肌损伤。 |
| 英文摘要: |
| ABSTRACT Objective: To observe and explore the effect of micro ribonucleic acid-21 (miR-21) on the Toll-like receptor 4 (TLR4)/nuclear factor-κB (NF-κB) pathway in cardiomyocytes of rats with myocardial ischemia. Methods: Forty SD rats were randomly divided into sham operation group (10 rats) and modeling group (30 rats). The rats in the modeling group were subjected to ligation of the left anterior descending coronary artery (LAD) to establish a rat model of myocardial ischemia. Rats in the sham operation group were sutured without any other treatment after thoracotomy. Rats with successful modeling were randomly divided into control group (transfected with miR-NC lentivirus), silent group (transfected with miR-21 antagomir lentivirus), and overexpression group (transfected with miR-21 mimics lentivirus). The sham operation group was injected with the same volume of normal saline. Hematoxylin-eosin (HE) staining was used to observe myocardial tissue damage. TdT-mediated dUTP nick end labeling (TUNEL) method was used to detect cell apoptosis rate. Real-time quantitative polymerase chain reaction (RT-qPCR) and western blotting (WB) were used to detect the TLR-4, NF-κB, Caspase 3, B-lymphoma-2 gene (Bcl-2), Bcl-2-associated X protein (Bax) messenger ribonucleic acid (mRNA) and protein expressions, as well as phosphorylated NF-κB p65(p-NF-κB p65) level. Results: The myocardial fibers in the control group were disordered, and a large number of myocardial cells were swollen and necrotic, accompanied by inflammatory cell infiltration. The myocardial tissue damage in the silent group was further aggravated, while the myocardial tissue damage in the overexpression group was significantly improved. Compared with the sham operation group, the myocardial cell apoptosis rate, TLR-4, NF-κB, Caspase-3, Bax mRNA and their protein expressions and p-NF-κB p65 levels were increased in the control group, the silence group, and the overexpression group, while the Bcl-2 mRNA and its protein expression were decreased (P<0.05). Compared with the control group, the myocardial cell apoptosis rate, the mRNA and protein expressions of TLR-4, NF-κB, Caspase-3, Bax, and the level of p-NF-κB p65 in the silence group were increased, while the Bcl-2 mRNA and its protein expression were decreased(P<0.05), and the myocardial cell apoptosis rate, the mRNA and protein expressions of TLR-4, NF-κB, Caspase-3, Bax, and the level of p-NF-κB p65 in the overexpression group were decreased, while the Bcl-2 mRNA and its protein expression were increased(P<0.05). Conclusion: Down-regulation of miR-21 expression can promote the expression of TLR-4/NF-κB pathway and aggravate myocardial injury in rats with myocardial ischemia. |
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