〔Abstract〕 Objective To investigate the effects of sulforaphane(SFN) in regulating the macrophage glycolysisviathe arachidonate 5-lipoxygenase(ALOX5)/nuclear factor kappa B(NF-κB) signaling pathway on the progression of diabetic nephropathy(DN). Methods Bioinformatics analysis was used to identify the target genes of SFN in the treatment of DN. Human proximal tubular epithelial cell line(HK-2 cells) was induced with 30 mmol/L high glucose(HG) to create an in vitro model of DN. HK-2 cells were divided into the following groups: normal glucose(NG) group, HG group, HG+SFN(3 mmol/L) group, HG+ALOX5 group, HG+SFN(3 mmol/L) +ALOX5 group, HG-treated macrophages+HK-2 group, HG+SFN(3 mmol/L) treated macrophages s+HK-2 group, HG+ ALOX5 transfection treated macrophages+HK-2 group, HG+SFN(3 mmol/L) + ALOX5 transfection treated macrophages+HK-2 group. CCK-8 assay was used to detect cell viability, Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling(TUNEL) method was used to detect cell apoptosis; glucose and lactate levels in the cells were measured using assay kits; Western blot was performed to detect the expression of ALOX5, NF-κB, and glycolysis-related proteins hexokinase-2(HK2), pyruvate kinase M2(PKM2), glucose transporter 1(GLUT1) in each group. Diabetic nephropathy(DN) mouse models were established using streptozotocin(STZ) and treated with SFN(0.5 mg/kg). Various biochemical parameters were measured in the mice, and kidney tissue pathology was examined using H&E staining. Western blot was used to detect the expression of glycolysis-related proteins(HK2, PKM2, GLUT1) in kidney macrophages. Results Bioinformatics analysis revealedALOX5as the target gene of SFN in treating DN. Compared to the HG group, SFN treatment enhanced HK-2 cell viability and inhibited apoptosis(P<0.05); concurrently, SFN treatment suppressed HG-induced macrophage glycolysis-related protein and attenuated macrophage-mediated HK-2 cellular injury(P<0.05). Western blot results showed that SFN inhibited the expression of ALOX5 and NF-κB(P<0.05). The mouse experiment results showed that SFN treatment improved kidney function and pathological changes in the kidney of DN mice, and inhibited the related protein expression of acrophage glycolysis in kidney tissue(P<0.05). Conclusion SFN improves the progression of DN by inhibiting the expression of macrophage glycolysis-related protein through theALOX5/NF-κB signaling pathway.