The anti-obesity effects of walnut meal enzymatic digests were investigated using a C57BL/6 mouse model of high-fat diet (HFD)-induced obesity and a differentiated 3T3-L1 adipocyte model. The accumulation of lipids in the adipocytes was identified through the use of oil red O fat staining. The levels of triglycerides (TG)， glycerol (Gly)， and free fatty acids (FFA) within the cells were quantified using a commercially available kit. The expression of genes involved in fat synthesis was determined through real-time fluorescence quantitative polymerase chain reaction (PCR RT-qPCR). The results demonstrated that the enzymatic digestion of walnut meal markedly diminished the accumulation of lipid droplets in adipocytes， notably reduced the TG and FFA content in adipocytes， and enhanced the Gly content in the cell supernatant. Concurrently， the enzymatic digestion of walnut meal significantly diminished the mRNA expression levels of adipose synthesis-related genes. The levels of free fatty acids (FFA) decreased by 40.14% and 39.50%， respectively， while the level of glycine increased by 35.12%. Additionally， the expression levels of mRNA for genes PPAR-γ and C/EBPα in adipocytes were found to be decreased by 31.42% and 55.25%， respectively， when the mass concentration of the enzymatic digest of walnut meal reached 50 μg/mL. Furthermore， the enzymatic digest of walnut meal was observed to significantly reduce body mass， adipose tissue mass and food intake in mice fed a high-fat diet (HFD). The study demonstrated that the enzymatic digest of walnut meal inhibited lipid accumulation in 3T3-L1 adipocytes， and its mechanism of action was related to the inhibition of the expression of genes related to lipid synthesis. Additionally， it was found to inhibit lipid accumulation in HFD.