Zedoarondiol Targets GYG1 to Alleviate Myocardial Ischemia-Reperfusion Injury

Myocardial ischemia/reperfusion injury (MIRI) remains a major clinical problem, marked by acute bioenergetic dysfunction and Ca2+ overload that contribute to cardiomyocyte death after ischemia. In this study, we showed that Zedoarondiol (Zed), a natural compound, significantly attenuated myocardial injury and cardiomyocyte death in both a rat MIRI model in vivo and an oxygen-glucose deprivation/reoxygenation (OGD/R) cardiomyocyte model in vitro. By integrating reverse virtual screening, molecular docking, and molecular dynamics simulations, we identified GYG1 as a putative binding target of Zed. Surface plasmon resonance (SPR) analyses showed concentration-dependent binding curves, confirming the physical interaction. Importantly, siRNA-mediated knockdown of GYG1 markedly abolished the cardioprotective effects of Zed. Furthermore, analysis of publicly available single-nucleus RNA-sequencing (snRNA-seq) data (GEO: GSE240848), combined with in silico virtual-knockout analysis, revealed that Gyg1 deletion significantly disrupts key glycogen metabolism and Ca2+ handling pathways in cardiomyocytes. Cellular assays were in line with these findings and showed that Zed restored OGD/R-induced intracellular Ca2+ homeostasis in a GYG1-dependent manner. Collectively, these results support GYG1 as a critical functional target of Zed and highlight Zed as a promising small-molecule candidate for the treatment of MIRI.