Salvianic Acid A Regulates Ferroptosis by Activating the Nrf2-GPX4 Pathway Through EGFR to Protect Myocardial Ischemia-Reperfusion Injury

Objective: Salvianic acid A, known for its potent cardioprotective effects, is a promising therapeutic agent for myocardial ischemia-reperfusion (I/R) injury, a condition where ferroptosis plays a critical role. This study aims to explore the effect of Salvianic acid A on I/R injury through EGFR-Nrf2-GPX4-mediated ferroptosis. Methods: The levels of ROS, GSH, GSSG, Fe 2+ , the state of ferroptosis, the expression and localization of Nrf2, and the expression of EGFR were all detected in hypoxia/reoxygenation (H/R) cells. The Nrf2 inhibitor ML385 and siRNA-EGFR were used to confirm the involvement of EGFR/Nrf2 in protecting cardiomyocytes from H/R-induced ferroptosis. Additionally, the regulatory effect of Salvianic acid A on EGFR/Nrf2/GPX4 were analyzed in the I/R animal model with EGFR inhibitors/agonists. Results: Salvianic acid A inhibited ROS, Fe 2+ , GSH, GSSG, GSH/GSSG, and promoted GPX4 and SLC7A11 expressions in H/R cells. Further experiments demonstrated that Salvianic acid A promoted Nrf2 nuclear translocation and expression to inhibit ferroptosis, which was abolished by the Nrf2 inhibitor ML385. In addition, the molecular docking indicated that Salvianic acid A has a good binding activity with EGFR, and cell experiments also confirmed that Salvianic acid A promoted EGFR expression, which was required for its subsequent activation of Nrf2 and inhibition of ferroptosis, as verified by siRNA-EGFR. In vivo experiments showed that Salvianic acid A alleviated myocardial injury, inhibited ferroptosis, and this improvement was related to the regulation of NRF2-GPX4 by EGFR. Conclusion: Salvianic acid A mediated ferroptosis through the EGFR-regulated NRF2-GPX4 pathway, ultimately improving myocardial I/R injury. Salvianic acid A protects heart from injury by regulating EGFR, Nrf2 and GPX4.The image illustrates a biochemical pathway titled ’Myocardial ischemia-reperfusion injury’. It depicts salvianic acid A protecting against hypoxia and stress-induced injury. Salvianic acid A interacts with EGFR, leading to activation of Nrf2. Nrf2 translocates to the nucleus, where it binds to ARE, promoting gene expression. In the cytoplasm, GPX4 is activated, inhibiting lipid peroxidation and reducing the labile iron pool, preventing mitochondrial dysfunction. This process inhibits ferroptosis and myocardial ischemia-reperfusion injury. The pathway highlights interactions between EGFR, Nrf2, Keap1, GPX4 and glutathione, with arrows indicating activation and inhibition steps.