摘要Objective This study aimed to investigate the protective effects of berberine(BBR)on pancreatic β-cells and explore its underlying molecular mechanisms via a proteomics-based approach.Methods Using db/db mice as a diabetes model,BBR was administered at doses of 100 mg/kg and 200 mg/kg for 8 weeks.The protective effects were assessed through fasting blood glucose(FBG),oral glucose tolerance test(OGTT),insulin tolerance test(ITT),pancreatic histopathological analysis,and TUNEL staining.Proteomic analysis employing the data-independent acquisition(DIA)method identified differentially expressed proteins(DEPs),whereas Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses were conducted to identify poten-tial pathways.Molecular docking,surface plasmon resonance(SPR),and immunohistochemistry(IHC)were performed to validate key target proteins.Results BBR significantly reduced blood glucose levels,improved insulin resistance,enhanced insulin secretion,and reversed pathological changes in pancreatic tissue,thereby alleviating β-cell damage.Proteomic analysis identified 171 DEPs,impli-cating the AGE/RAGE signaling pathway as a key mechanism through which BBR exerts its protective effects.The results of molecular docking,SPR and IHC confirmed that BBR markedly inhibited the activation of the AGE/RAGE pathway.Conclusions These findings suggest that BBR alleviates pancreatic β-cell damage,potentially through regulation of the AGE/RAGE pathway,providing insights into its therapeutic potential for diabetes management.