Dapagliflozin Ameliorates Myocardial Ischemia/Reperfusion Injury by Modulating EGFR Signaling and Targeting NCOA4-mediated Ferritinophagy

SGLT2 inhibitor dapagliflozin (Dapa) has gained increasing attention in the treatment of myocardial ischemia-reperfusion injury (IRI). However, the mechanism of action of the cardiovascular benefits of Dapa is unclear. The present study aimed to investigate the effects of Dapa on myocardial IRI and the underlying molecular mechanisms. The effects of Dapa on myocardial IRI were investigated using the in vitro perfusion Langendorf model and the in vitro hypoxia/reoxygenation (H/R) cell model. Histological changes, myocardial enzymes, oxidative stress and mitochondrial structure/function were assessed. Mechanistic studies involved various molecular biology methods such as ELISA, immunoprecipitation, western blot, immunofluorescence and Bioinformatics. Our findings demonstrate that Dapa upregulates EGFR phosphorylation, suppresses NHE1 expression in myocardial tissues, modulates NCOA4-mediated ferritinophagy to enhance mitochondrial function, reduces ROS expression, and mitigates myocardial IRI. In the Langendorf model, Dapa effectively attenuates cardiac dysfunction, myocardial injury, mitochondrial damage, and oxidative imbalance induced by ischemia-reperfusion. In vitro experiments revealed that blocking EGFR or autophagy with inhibitors (AG and Baf, respectively) or inducing ferroptosis with Era promotes ROS release, exacerbates mitochondrial injury, and diminishes the protective effects of Dapa. Notably, Era did not affect NCOA4-mediated ferritinophagy. Conversely, the EGFR agonist NSC counteracted these effects, underscoring that Dapa confers cardioprotection by modulating mitochondrial function through EGFR-mediated regulation of NCOA4-mediated ferritinophagy. In summary, Dapa activates EGFR phosphorylation, regulates NCOA4-mediated ferritinophagy, modulates mitochondrial function, and effectively mitigates myocardial IRI. These findings provide a robust theoretical foundation for the clinical application of Dapa in treating cardiovascular conditions.