Institute of Hypertension Vascular Disease, Guangxi International Zhuang Medicine Hospital, Guangxi University of Chinese Medicine, No.8 Qiuyue Road, Wuxiang New District, Nanning, 530201, Guangxi, China.
Cardiology Department, Guangxi International Zhuang Medicine Hospital, Guangxi University of Chinese Medicine, Nanning, 530201, China.
Hum Cell. 2021 Mar;34(2):360-371. doi: 10.1007/s13577-020-00482-z. Epub 2021 Jan 15.
Myocardial ischemia/reperfusion injury (MIRI) is a major cause of acute cardiac injury that is associated with high morbidity and mortality, and for which specific treatments are lacking. In this study, we investigated the underlying molecular mechanism of miR-144-3p in the pathological process of MIRI. A mouse I/R injury model and H9c2 cardiomyocyte hypoxia/reoxygenation (H/R) model were used to simulate the ischemia/reperfusion process in vivo and in vitro, respectively, and the relative expression and regulatory effect of miR-144-3p were determined. The target of miR-144-3p was also verified by a luciferase reporter assay. We found that miR-144-3p was significantly downregulated in mouse myocardium subjected to I/R and cardiomyocytes subjected to H/R. Upregulation of miR-144-3p significantly attenuated MIRI in vivo and in vitro. A Ca-activated chloride channel-TMEM16A (ANO1)-was identified as a target gene of miR-144-3p through bioinformatic analysis. The interaction between miR-144-3p and the 3'-untranslated region of ANO1 was confirmed with dual-luciferase reporter assay, RNA immunoprecipitation assay, real-time quantitative polymerase chain reaction, and western blot analysis. Moreover, by targeting ANO1, miR-144-3p inhibited the activation of NLRP3 inflammasome inflammatory signals in myocardial cells. Collectively, the present study provides a novel insight into the role of miR-144-3p in the inhibition of MIRI, suggesting that the miR-144-3p/ANO1 axis may be a putative therapeutic target in myocardial ischemia.
心肌缺血/再灌注损伤（MIRI）是一种主要的急性心脏损伤原因，与高发病率和死亡率相关，目前缺乏特定的治疗方法。在本研究中，我们研究了 miR-144-3p 在 MIRI 病理过程中的潜在分子机制。使用小鼠 I/R 损伤模型和 H9c2 心肌细胞缺氧/复氧（H/R）模型分别在体内和体外模拟缺血/再灌注过程，确定了 miR-144-3p 的相对表达和调节作用。还通过荧光素酶报告基因检测验证了 miR-144-3p 的靶标。我们发现，miR-144-3p 在经历 I/R 的小鼠心肌中和经历 H/R 的心肌细胞中均显著下调。miR-144-3p 的上调显著减轻了体内和体外的 MIRI。通过生物信息学分析，鉴定出 Ca2+激活的氯离子通道-TMEM16A（ANO1）是 miR-144-3p 的靶基因。通过双荧光素酶报告基因检测、RNA 免疫沉淀实验、实时定量聚合酶链反应和 Western blot 分析证实了 miR-144-3p 与 ANO1 的 3'-非翻译区的相互作用。此外，通过靶向 ANO1，miR-144-3p 抑制了心肌细胞中 NLRP3 炎性小体炎症信号的激活。总之，本研究为 miR-144-3p 在抑制 MIRI 中的作用提供了新的见解，表明 miR-144-3p/ANO1 轴可能是心肌缺血的潜在治疗靶点。
