Diabetic nephropathy (DN) is the most important complication in patients with diabetes. The accumulation of advanced glycation end-products (AGEs) is the main reason for the development of DN. In this study, we investigated the mechanism of buckwheat hull flavonoids to break AGEs in vitro by measuring fluorescence analysis, three-dimensional fluorescence, protein molecular weight, free amino groups, and the sulfhydryl group content. Proteomics analysis was used to determine the effect of total buckwheat hull flavonoids (TBHF) intervention on protein differential expression in the kidney of db/db mice. The results showed that buckwheat hull flavonoids were potent in breaking AGEs in vitro, and they protected mice kidneys by regulating the renal AGE-RAGE pathway. This study lays a strong experimental and theoretical foundation for the development of new lysing agents to break AGEs. The findings should make an important contribution to the field of flavonoids in improving the application of diabetic nephropathy in the diet.