Dapagliflozin attenuates cardiomyocyte hypertrophy by inhibiting KEAP1-NRF2-mediated ferroptosis pathway.
Abstract
Objectives
Myocardial hypertrophy is a common pathological process in various cardiovascular diseases and may ultimately lead to heart failure. As a sodium-glucose cotransporter 2 inhibitor, dapagliflozin (DAPA) has been demonstrated to be cardioprotective, in addition to its hypoglycemic effect. However, the precise mechanism of DAPA in myocardial hypertrophy remains unclear.
Methods
H9C2 rat cardiomyoblast cell line were subjected to isoproterenol (ISO)-induced hypertrophy and erastin-induced ferroptosis, and treated with DAPA, ferrostatin-1, KEAP1 activator K67, or inhibitor ML334. Hypertrophic markers and cell surface area were assessed by quantitative real-time polymerase chain reaction and rhodamine-phalloidin staining. Ferroptosis was evaluated by measuring glutathione (GSH), malondialdehyde, reactive oxygen species, lipid peroxidation, intracellular ferrous iron, and mitochondrial membrane potential. Protein expression of KEAP1, NRF2, solute carrier family 7 member 11, and GSH peroxidase 4 was determined by western blot. KEY
Findings
DAPA attenuated ISO-induced cardiomyocyte hypertrophy and suppressed ferroptosis by inhibiting KEAP1 to activate NRF2, upregulating downstream targets. GSH peroxidase 4 knockdown abolished DAPA's protective effects. KEAP1 reactivation with K67 reversed DAPA-mediated cardioprotection, whereas KEAP1 inhibition with ML334 recapitulated it.
Conclusions
DAPA exerts a protective role in myocardial hypertrophy by inhibiting KEAP1-NRF2-mediated ferroptosis pathway, providing novel insights into the potential mechanism and application of DAPA on cardiovascular diseases.