Dapagliflozin improves diabetic nephropathy by regulating autophagy and ferroptosis through CYP450s/ROS pathway

Diabetic nephropathy (DN) is a severe complication of diabetes, presenting significant clinical challenges due to its refractory nature. Although dapagliflozin is used as a clinical intervention, its precise molecular mechanisms remain incompletely understood. Our study employed an integrative approach combining bioinformatics analysis with experimental validation to systematically investigate dapagliflozin's therapeutic potential in DN. Through comprehensive bioinformatics screening, we identified cytochrome P450 (CYP450) enzymes as key molecular targets in DN pathogenesis, a finding subsequently validated using Nephroseq database analysis. A DN model was established in db/db mice, and dapagliflozin was administered to investigate its molecular mechanisms. Results showed that DN is closely associated with cytochrome P450 metabolism pathways, autophagy and oxidative stress. Dapagliflozin could suppress the upregulated expression of CYP1A1 and CYP1B1. Importantly, dapagliflozin treatment effectively reduced oxidative stress and enhanced cellular autophagic activity, as evidenced by increased expression of Beclin-1, LC3II, and ATG7 coupled with decreased p62 levels. Furthermore, dapagliflozin decreased ferroptosis by raising GPX4 levels and lowering Fe²⁺ levels. Additionally, dapagliflozin reduced NCOA4 and TFR1 expression while increasing FTH1, FTL1, and FPN1 expression. These findings provide compelling evidence that dapagliflozin's renoprotective effects involve suppression of NCOA4-mediated ferroptosis and coordinated regulation of CYP450 metabolism, autophagy, and oxidative stress pathways. Our study not only elucidates the multifaceted mechanisms underlying dapagliflozin's therapeutic efficacy in DN but also identifies new potential targets for DN treatment strategies.