Vaccarin Improves Myocardial Ischemia-Reperfusion Injury by Attenuating Oxidative Stress and Ferroptosis Through Reducing NOX4-Modulated JAK2/STAT3 Pathway Activation

Vaccarin is a naturally occurring flavonoid glycoside with cardioprotective properties; however, its involvement in myocardial ischemia-reperfusion (MI/R) injury remains poorly defined. We herein investigated whether vaccarin protects cardiomyocytes from MI/R-induced dysfunction in vivo and in vitro and examined the associated molecular mechanisms. To explore the protective effects of vaccarin, we established an MI/R injury mouse model and in vitro hypoxia/reoxygenation (H/R) injury model using H9c2 cardiomyocytes. In vivo results showed that vaccarin alleviated MI/R injury. In H9c2 cells, vaccarin partially reversed the H/R-induced elevation of Reactive Oxygen Species and malondialdehyde levels and restored the reduced activity of superoxide dismutase. Vaccarin also suppressed the H/R-induced accumulation of Fe²⁺ and glutathione disulfide while restoring the levels of glutathione, solute carrier family 7 member 11, and Glutathione Peroxidase 4. NADPH Oxidase 4 (NOX4) was subsequently identified as a putative molecular target of vaccarin. Moreover, vaccarin inhibited H/R-induced activation of the Janus kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) signaling pathway in cardiomyocytes. NOX4 overexpression counteracted vaccarin-mediated suppression of JAK2/STAT3 signaling and blocked its antioxidative and antiferroptotic effects in H/R-treated cardiomyocytes. Collectively, these findings indicate that vaccarin confers cardioprotection against MI/R injury by suppressing oxidative stress and Ferroptosis, at least partly through inhibition of NOX4-driven JAK2/STAT3 signaling. These results suggest that vaccarin may be a promising therapeutic candidate for the treatment of MI/R injury.