Reynoutrin activates Sirt1/Foxo3a and Nrf2 pathway via targeting Fyn to ameliorate diabetic nephropathy

BACKGROUND: Diabetic nephropathy (DN) is a major complication of diabetes, associated with high mortality and disability rates. Fyn kinase has emerged as a promising therapeutic target for DN due to its antioxidant properties. However, effective therapeutic agents specifically targeting Fyn are still lacking. OBJECTIVE: This study aimed to identify a novel natural Fyn inhibitor and explore its therapeutic effects and mechanisms in DN. METHODS: A structure-based virtual screening of 60,556 natural compounds was conducted to discover Fyn-targeting molecules. The interaction between the compound and Fyn was validated by drug affinity responsive target stability, differential scanning fluorimetry, surface plasmon resonance, molecular docking, and cellular thermal shift assay. Therapeutic efficacy was assessed in streptozotocin (STZ) combined high-fat-fed diabetic mice and high glucose-stimulated glomerular mesangial cells. Non-targeted metabolomics was used to explore potential mechanisms. RESULTS: We identified reynoutrin, a natural small molecule, as a potent inhibitor of Fyn from a library of 60,556 natural compounds. Reynoutrin directly interacted with the Fyn protein, forming five key hydrogen bonds that inhibited both the expression and phosphorylation of Fyn. Non-targeted metabolomics linked reynoutrin's effects to the Foxo signaling pathway and oxidative phosphorylation. Mechanistically, reynoutrin activated the downstream Sirt1/Foxo3a and Nrf2 antioxidant signaling pathways by inhibiting Fyn, thereby enhancing the activity of antioxidant enzymes and alleviating diabetic renal injury. CONCLUSION: Our results emphasize the efficacy of reynoutrin in treating DN and demonstrate its broader therapeutic potential through Fyn inhibition.