Dapagliflozin confers protection against ferroptosis in cardiomyocyte via the inhibition of interferon-gamma pathway

BACKGROUND: Ferroptosis is reported to have a role in cardiovascular disease (CVD) progression. Recent emerging evidences demonstrate the role of Sodium-Glucose Co-Transporter 2 (SGLT-2) inhibitors in CVD owing to its promising cardioprotective benefits. However, the detailed molecular mechanism underlying its cardioprotective effects remain incompletely understood. We aimed to investigate the potential ferroptosis-inhibitory mechanism of SGLT-2 inhibitor in HL-1 mouse cardiomyocytes. METHODS AND RESULTS: ) to simulate ischemic injury with or without dapagliflozin (DAPA, 10 µM), a SGLT-2 inhibitor. Reactive oxygen species (ROS) was detected by immunofluorescence. Ferroptosis activity was analysed by measuring levels of intracellular iron, lipid peroxidation, malondialdehyde (MDA), and glutathione (GSH). Increase in ROS generation, intracellular iron, MDA concentration, and lipid peroxidation indicated increasing ferroptosis activity during hypoxic stress in cardiomyocytes. Treatment with DAPA caused reduction in ferroptosis. Pathway-specific gene analysis showed that interferon-gamma (IFN-γ) was upregulated during hypoxia, and was downregulated with DAPA treatment. Silencing of IFN-γ by using small interfering RNA (siRNA) resulted in inhibition of lipid peroxidation; on the other hand, incorporating recombinant IFN-γ resulted in the reversal of this phenomenon, thus validating the role of IFN-γ in hypoxia-induced ferroptosis. CONCLUSION: Our findings reveal IFN-γ pathway as a novel intracellular target of ferroptosis inhibition by DAPA in cardiomyocytes, providing molecular insight into the mechanisms underlying the cardioprotective effects of SGLT-2 inhibitor.