This study evaluated the effect on Wistar rat's oxidative status of incorporating amaranth flour (AF) and protein isolate (AI) in increased-fat diets. Five of the groups were fed for 4 weeks with either BD (basal diet), Chol+F (2% cholesterol, 10% porcine fat), Chol+F+E (0.005% α-tocopherol), Chol+F+AF 1 or Chol+F+AI 1 (25% of protein replacement) diets. The other two groups were fed for 4 weeks with Chol+F and then 1 week with Chol+F+AF 2 or Chol+F+AI 2 diet (50% of protein replacement). Various effects on the oxidative stress biomarkers in tissues (intestine and liver) were observed. These effects were dependent on the ingredients, dose, and administration time. In the intestinal cells, Chol+F+AF 1 and Chol+F+AI 2 produced an increment in the reduced glutathione (GSH) content (56% and 39%, respectively), while Chol+F+AF 2 induced an increment in the superoxide dismutase (SOD) (25%) and glutathione peroxidase (GPx) (46%) activities. The presence of certain components in flour (e.g., fiber, polyphenols, squalene) could explain the higher activity recorded for AF. In the liver, Chol+F+AF 2 produced a decrease in SOD (19%) and GSH (36%), as well as an increase in GPx (255%); Chol+F+AI 1 and Chol+F+AI 2 also produced a decrease in GSH (36% and 24%, respectively) and important increments in GPx activity (273% for Chol+F+AI 1 and 2,900% for Chol+F+AI 2). These effects were dependent on the AI dose and were probably produced by absorbed peptides. a Milk casein, 92% w/w protein (Droguería Industrial Lanús S.A., Argentina). b Dextrin (Cofem S.A., Argentina). c Antioxidants-free soy oil