Ilex Pubescens Inhibits Ferroptosis Post-Myocardial Ischemia-Reperfusion Injury by Activating Aldehyde Dehydrogenase 2

Background Ilex pubescens, a traditional Chinese herb widely used in cardiovascular diseases, exhibited potential anti-inflammatory capabilities in myocardial ischemia-reperfusion injury (MI/RI). Aldehyde dehydrogenase 2 (ALDH2) plays a crucial role in the cascade of events leading to ferroptosis, a form of controlled cellular demise involving iron-dependent lipid peroxidation and associated with MI/RI. However, the impact of Ilex pubescens on MI/RI remains unclear.Purpose This work aimed to elucidate the primary mechanism by which Ilex pubescens (IPES) extracts reduce ferroptosis post-MI/RI.M ethodsAn in vivo ischemia-reperfusion (I/R) model was established by occluding the left anterior descending coronary artery (LAD) in C57BL/6 mice for 30 minutes, followed by 24 hours of reperfusion. To induce ferroptosis in vitro, primary neonatal cardiomyocytes, extracted from Sprague-Dawley rats, were stimulated with erastin, an inducer of ferroptotic cell death. Furthermore, the selective ALDH2 inhibitor CVT-10216 was employed to confirm that IPES reduces ferroptosis in myocardial cells through ALDH2 activation.Results IPES exerted significant cardioprotective effects. In the mouse I/RI model group, IPES significantly reduced cardiac injury, improved cardiac function, and lowered serum biomarkers of cardiac damage. Furthermore, treatment with IPES significantly reduced inflammatory cell infiltration, myocardial intracellular ROS generation, and iron deposition-induced damage. Furthermore, it significantly decreased the expression levels of iron-related proteins transferrin receptor 1 (TFR1), ferritin, ferritin heavy chain 1 (FTH1), ferritin light chain 1 (FTL1), and the concentration of the ferroptosis-associated metabolite aldehyde 4-hydroxynonenal (4-HNE). IPES prevented the degradation of GSH while maintaining the activity of glutathione peroxidase 4 (GPX4). Importantly, IPES substantially increased the ALDH2 expression level, emphasizing its inhibitory effect on myocardial injury. IPES provided in vitro protection for primary cardiomyocytes against erastin-induced ferroptotic cell death. Specific inhibition of ALDH2 by CVT10216 partially attenuated the protective effects of IPES.Conclusion The results demonstrate that IPES inhibits ferroptosis post-MI/RI by activating ALDH2, strongly supporting its potential use in MI/RI.