The primary mechanism underlying insulin-resistant diabetes is the disruption of insulin signalling due to a relative deficiency of insulin receptor (InsR). EphB4, which forms a complex with InsR to promote lysosomal degradation, negatively regulates this pathway. Quercetin-3-O-β-D-pyranoside (Reynoutrin, Rey), a small natural compound, has garnered attention for its diverse biological activities. In this study, we demonstrated that Rey improved glycolipid metabolism in streptozotocin (STZ)-induced diabetic mice on a high-fat diet (HFD) and in palmitic acid (PA)-treated HepG2 cells. Furthermore, network pharmacology screening identified EphB4 as a potential target of Rey in the regulation of insulin resistance (IR). Surface plasmon resonance (SPR), drug affinity responsive target stability (DARTS), and cellular thermal shift assay (CETSA) results confirmed that Rey directly bond to EphB4. Notably, molecular docking and CETSA analyses revealed that Rey interacted with key amino acids in EphB4, including Phe759 and Met696, thereby spatially inhibiting the interaction between EphB4 and InsR to mechanically prevent the degradation of InsR, which contributed to improve IR. In summary, our study identified Rey as a promising drug candidate for diabetes treatment, directly targeting EphB4 to improve insulin resistance and glycolipid metabolism in Type 2 diabetes mellitus.
Keywords: EphB4; Glucose and lipid metabolic disorders; InsR; Insulin resistance; Reynoutrin; Type 2 diabetes mellitus.
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