miR-190-5p 通过靶向 PHLLP1 减轻心肌缺血再灌注损伤。
miR-190-5p Alleviates Myocardial Ischemia-Reperfusion Injury by Targeting PHLPP1.
机构信息
Department of Cardiology, The Second Hospital of Jilin University, Changchun, China.
Department of Cardiology, Beijing Tsinghua Changgung Hospital, Tsinghua University, Beijing, China.
出版信息
Dis Markers. 2021 Nov 25;2021:8709298. doi: 10.1155/2021/8709298. eCollection 2021.
OBJECTIVE
Myocardial ischemia-reperfusion (I/R) injury (MIRI) refers to the more serious myocardial injury after blood flow recovery, which seriously affects the prognosis of patients with ischemic cardiomyopathy. This study explored the new targets for MIRI treatment by investigating the effects of miR-190-5p and its downstream target on the structure and function of myocardial cells.
METHODS
We injected agomir miR-190-5p into the tail vein of rats to increase the expression of miR-190-5p in rat myocardial cells and made an I/R rat model by coronary artery occlusion. We used 2,3,5-triphenyl tetrazolium chloride staining, lactate dehydrogenase (LDH) detection, echocardiography, and hematoxylin-eosin (HE) staining to determine the degree of myocardial injury in I/R rats. In addition, we detected the expression of inflammatory factors and apoptosis-related molecules in rat serum and myocardial tissue to determine the level of inflammation and apoptosis in rat myocardium. Finally, we determined the downstream target of miR-190-5p by Targetscan system and dual luciferase reporter assay.
RESULTS
The expression of miR-190-5p in an I/R rat myocardium was significantly lower than that in normal rats. After treatment of I/R rats with agomir miR-190-5p, the ischemic area of rat myocardium and the concentration of LDH decreased. The results of echocardiography and HE staining also found that overexpression of miR-190-5p improved the structure and function of rat myocardium. miR-190-5p was also found to improve the viability of H9c2 cells and reduce the level of apoptosis of H9c2 cells. The results of Targetscan system and dual luciferase reporter assay found that miR-190-5p targeted to inhibit pleckstrin homology domain leucine-rich repeat protein phosphatase 1 (PHLPP1). In addition, inhibition of PHLPP1 was found to improve the viability of H9c2 cells.
CONCLUSION
Therefore, miR-190-5p can reduce the inflammation and apoptosis of myocardium by targeting PHLPP1, thereby alleviating MIRI.
目的
心肌缺血再灌注（I/R）损伤（MIRI）是指血流恢复后更严重的心肌损伤，严重影响缺血性心肌病患者的预后。本研究通过研究 miR-190-5p 及其下游靶标对心肌细胞结构和功能的影响，探讨 MIRI 治疗的新靶点。
方法
我们通过尾静脉注射 agomir miR-190-5p 增加大鼠心肌细胞中 miR-190-5p 的表达，并通过冠状动脉阻塞制作 I/R 大鼠模型。我们使用 2,3,5-三苯基氯化四氮唑染色、乳酸脱氢酶（LDH）检测、超声心动图和苏木精-伊红（HE）染色来确定 I/R 大鼠的心肌损伤程度。此外，我们检测了大鼠血清和心肌组织中炎症因子和凋亡相关分子的表达，以确定大鼠心肌的炎症和凋亡水平。最后，我们通过 Targetscan 系统和双荧光素酶报告基因检测确定了 miR-190-5p 的下游靶标。
结果
I/R 大鼠心肌组织中 miR-190-5p 的表达明显低于正常大鼠。用 agomir miR-190-5p 处理 I/R 大鼠后，大鼠心肌缺血面积和 LDH 浓度降低。超声心动图和 HE 染色的结果也发现，过表达 miR-190-5p 改善了大鼠心肌的结构和功能。还发现 miR-190-5p 提高了 H9c2 细胞的活力，减少了 H9c2 细胞的凋亡水平。Targetscan 系统和双荧光素酶报告基因检测结果发现，miR-190-5p 靶向抑制pleckstrin 同源结构域富含亮氨酸重复蛋白磷酸酶 1（PHLPP1）。此外，抑制 PHLPP1 可提高 H9c2 细胞的活力。
结论
因此，miR-190-5p 通过靶向 PHLPP1 减轻 MIRI，从而减少心肌的炎症和凋亡。
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