Pumpkin (Cucurbita pepo L.) seeds are enriched in bioactive compounds having functional properties. The aim of this study was to analyze the pumpkin seed proteins (PSP) effects on insulin resistance, oxidative stress damage and inflammation in rats with high fructose-induced metabolic syndrome.Twenty four male Wistar rats, fed isoenergetic diets supplemented with: (1) 20% casein (C); (2) 20% casein and 1 g/kg/day PSP (P); (3) 20% casein and 64% D-fructose (C-HF); (4) 20% casein, 1 g/kg/day PSP and 64% D-fructose (P-HF). After 8 weeks of treatment, fructose supply impaired white adipose tissue (WAT) weight, deteriorated glucose tolerance and tAUC, plasma glucose, insulin, insulinogenic index, HOMA-IR and HOMA-β, antioxidant status, lipid and protein oxidation, plasma TNF-α and IL-6 as compared to control diets. Interestingly, rats assigned to the PSPs diet with or without fructose displayed lower plasma glucose, insulin and fructose, improved tolerance of glucose, tAUC, HOMA-IR and HOMA-β and increased insulinogenic index as compared to C diets. PSPs consumption lowered thiobarbituric acid reactive substances, hydroperoxides and carbonyls in WAT and carbonyls in muscle. Superoxide dismutase and glutathione peroxidase in WAT were significantly diminished in P-HF but increased in P as compared to C-HF and C. Rats fed P-HF diet had low catalase in WAT and high in muscle than those fed C-HF. Moreover, catalase activity increased in muscle but decreased in WAT in P group than in C group. In conclusion, pumpkin seed proteins exhibit favorable effects on metabolic disorders of fructose-induced metabolic syndrome, suggesting a key role in disease therapy.