Breviscapine attenuates myocardial ischemia-reperfusion injury by inhibiting ferroptosis via activation of NRF2/SLC7A11/GPX4 pathway

To explore the protective effect of Breviscapine against myocardial ischemia-reperfusion (I/R) injury and its mechanism. The in vivo and in vitro models of myocardial I/R injury were first established and treated with different concentrations of Bre. The myocardial infarct size was measured, while histopathological changes of myocardial tissue were evaluated by hematoxylin-eosin (HE) staining, and cardiac function-related parameters were assessed via echocardiography. Prussian blue staining was performed to visualize ferroptosis; enzyme-linked immunosorbent assay (ELISA) was used to detect the expression levels of creatine kinase-MB (CK-MB) and lactate dehydrogenase (LDH); flow cytometry was employed to determine the apoptosis rate of cardiomyocytes. Western blot (WB) and qRT-PCR were conducted to examine the levels of ferroptosis-related molecules (NOX1, NRF2, GPX4 and SLC7A11), respectively. Meanwhile, oxidative stress-related indices associated with ferroptosis, such as malondialdehyde (MDA), superoxide dismutase (SOD) and reactive oxygen species (ROS), were determined. Finally, a specific NRF2 inhibitor was administered for intervention, and changes in the above indicators were observed. Bre dose-dependently alleviated myocardial histological damage, reduced infarct size, improved cardiac function (elevated EF/FS, decreased LVEDV/LVESV), and inhibited cardiomyocyte apoptosis. Bre also suppressed ferroptosis, as shown by reduced Fe²⁺accumulation, MDA, GSSG, ROS, and NOX1 expression, and enhanced SOD activity, GSH content, and NRF2/SLC7A11/GPX4 expression. NRF2 inhibitor ML385 reversed these effects. Bre alleviates myocardial I/R injury by inhibiting ferroptosis via activating the NRF2/SLC7A11/GPX4 pathway.