Matrine protects against myocardial ischemia-reperfusion injury by suppressing ferroptosis through BTF3/EPHB2 signaling

Objective In this in-vitro study, by using H9C2 cardiomyocytes subjected to oxygen–glucose deprivation/reoxygenation (OGD/R), we evaluated the cardioprotective effects of matrine across graded concentrations and explored underlying mechanisms related to ferroptosis and BTF3/EPHB2 signaling. Materials and methods H9C2 cardiomyocytes were stochastically allocated into five groups: Control group (routine culture), OGD/R group (myocardial ischemia-reperfusion injury model), OGD/R + L-MAT group (30 μM MAT), OGD/R + M-MAT group (60 μM MAT), and OGD/R + H-MAT group (100 μM MAT). Cellular proliferative capacity was measured by the Cell Counting Kit-8 (CCK-8) assay. Oxidative stress markers (ROS, LDH, MDA, GSH) and ferroptosis-related proteins (ACSL4, FDX1, GPX4, FTL), as well as BTF3, EPHB2, p38, and p p38, were assessed by fluorescence imaging, ELISA, and Western blotting. Results Relative to the Control group, the OGD/R group exhibited a marked reduction in cellular proliferative capacity, a marked surge in both the rate of apoptotic cell death and the intracellular accumulation of ROS, coupled with significantly heightened concentrations of LDH and MDA alongside a discernible depletion of GSH content. Concurrently, the relative protein expression levels of ACSL4, FDX1, FTL, BTF3, EPHB2, and p-p38 were observed to be robustly upregulated, whereas the protein level of GPX4 exhibited a noticeable downregulation (all P < 0.05). Conversely, when contrasted with the OGD/R group, the OGD/R + L-MAT, OGD/R + M-MAT, and OGD/R + H-MAT groups displayed a distinct augmentation in cellular proliferative activity, a prominent reduction in the apoptotic index and intracellular ROS generation, accompanied by decreased concentrations of LDH and MDA as well as a restored elevation of GSH levels. Moreover, the relative protein abundances of ACSL4, FDX1, FTL, BTF3, EPHB2, and p-p38 were significantly diminished, while that of GPX4 was prominently increased (all P < 0.05). Conclusion MAT markedly attenuates OGD/R induced damage in H9C2 cardiomyocytes; these effects are associated with reduced oxidative stress, apoptosis, and ferroptosis-related protein changes and with downregulation of BTF3/EPHB2 signaling, suggesting, but not definitively proving, that this pathway may contribute to MAT's cardioprotective actions.