The production and accumulation of reactive oxygen species (ROS) may contribute to the development of insulin resistance, which is the vital feature of type 2 diabetes. Thioredoxin and glutathione are the major thiol antioxidants protecting cells from ROS-induced cytotoxicity. In this study, carmustine (BCNU) was used to specifically inhibit glutathione reductase and auranofin (AF) was used to specifically inhibit thioredoxin reductase. In assay1, BCNU was used to stimulate 3T3L1 adipocytes for 2h and 24h. Glucose uptake was significantly inhibited by 2h-long stimulation, but not affected by 24h-long stimulation. In assay2, BCNU and AF were used together to stimulate 3T3L1 adipocytes for 2h and 24h. Glucose uptake was dramatically inhibited by both 2h- and 24h-long stimulation. Using SILAC-based quantitative proteomics, we quantified 4533 and 4111 proteins respectively. Based on fuzzy c-means analysis, these proteins were clustered into eight groups, and some of them were tightly associated with glucose uptake changes. In BCNU-stimulation assay, proteins with negative glucose uptake correlation were mainly located in mitochrodria, enriched in oxidative phosphorylation pathway. However, proteins with positive glucose uptake correlation were mainly enriched in Proteasome. In BCNU/AF-stimulation assay, proteins with both negative and positive glucose uptake correlation were mainly located in mitochrodria. The former were enriched in oxidative phosphorylation pathway, but the latter played key roles in generation of precursor metabolites and energy, enriched in glycolysis/gluconeogenesis pathway and some other metabolism-related pathways. Several dynamic key proteins and well-known phosphorylation sites perhaps involved in insulin resistance were validated by western blot.