Asperosaponin VI ameliorates acute kidney injury via restoring metabolic-oxidative homeostasis in NRF2 and PPARα dependent manners

Background Acute kidney injury (AKI) is a critical clinical syndrome with limited therapeutic options. Metabolic dysfunction and oxidative stress are central pathogenic mechanisms, yet therapies simultaneously targeting both processes remain underdeveloped. Methods We investigated the renoprotective effects of Asperosaponin VI (AVI) using hypoxia/reoxygenation(H/R)-injured tubular cells, patient-derived kidney organoids, and murine ischemia-reperfusion injury (IRI) and cisplatin-induced AKI models. Mechanistic studies employed RNA sequencing, molecular docking, surface plasmon resonance (SPR) and loss-of-function approaches with specific pathway inhibitors. Results AVI significantly attenuated tubular cell injury by suppressing apoptosis, reducing ROS generation, preserving mitochondrial function, and promoting mitophagy in vitro. In vivo , AVI treatment (20 mg/kg/day) markedly ameliorated IRI- and cisplatin-induced AKI, improving renal function, reducing tubular damage, and decreasing inflammation. Transcriptomic profiling identified PPAR and glutathione metabolism as key enriched pathways. Mechanistically, AVI functions as a dual agonist: it binds PPARα's ligand-binding domain (-8.1 kcal/mol; K D =0.815 μM), activating fatty acid oxidation genes (CPT1A, ACOX1) to restore energy metabolism and prevent lipotoxicity; simultaneously, it binds Keap1′s Kelch domain (-7.8 kcal/mol; K D =2.16 μM), disrupting Keap1-NRF2 interaction to activate antioxidant defenses (HO-1, NQO1) and preserve mitochondrial homeostasis. Critically, co-administration of PPARα antagonist GW6471 and NRF2 inhibitor ML385 abolished AVI's renoprotective effects, demonstrating that both pathways are indispensable for therapeutic efficacy. Conclusions AVI protects against AKI through synergistic activation of PPARα-mediated metabolic restoration and NRF2-mediated antioxidant defense. This dual-pathway mechanism represents a promising therapeutic strategy with strong translational potential for AKI management.