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 Ca²⁺-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 3ʹ-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) 是急性心脏损伤的主要原因，与高发病率和死亡率相关，并且缺乏具体的治疗方法。在这项研究中，我们研究了 miR-144-3p 在 MIRI 病理过程中的潜在分子机制。采用小鼠I/R损伤模型和H9c2心肌细胞缺氧/复氧（H/R）模型分别模拟体内和体外的缺血/再灌注过程，研究miR-144-3p的相对表达和调节作用。决定。miR-144-3p 的靶标也通过荧光素酶报告基因检测进行了验证。我们发现 miR-144-3p 在接受 I/R 的小鼠心肌和接受 H/R 的心肌细胞中显着下调。miR-144-3p 的上调显着减弱了体内和体外的 MIRI。钙²⁺激活的氯离子通道——TMEM16A (ANO1)——通过生物信息学分析被确定为 miR-144-3p 的靶基因。miR-144-3p 与 ANO1 的 3ʹ-非翻译区之间的相互作用通过双荧光素酶报告基因测定、RNA 免疫沉淀测定、实时定量聚合酶链反应和蛋白质印迹分析得到证实。此外，通过靶向 ANO1，miR-144-3p 抑制了心肌细胞中 NLRP3 炎性体炎症信号的激活。总的来说，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推定治疗靶点。