Effects of Different Doses of Excessive Iron in Diets on Oxidative Stress in Immune Organs of Sheep.

The aim of this work is to investigate the relationship between iron and oxidative stress in the immune organs of excessive iron-fed sheep. Sixteen German Mutton Merino rams were randomly divided into 4 groups, which were fed the basal diets supplemented with 50 (CON), 500 (L-iron), 1000 (M-iron), and 1500 (H-iron) mg Fe/kg as ferrous sulfate monohydrate (FeSO4·H2O), respectively. The actual iron content in the diet was determined to be 457.68 (CON), 816.42 (L-iron), 1256.78 (M-iron), and 1725.63 (H-iron) mg/kg, respectively. The consequences of oxidative damage were tested in 4 groups. The results showed that the contents of malondialdehyde (MDA), nitric oxide (NO), hydrogen peroxide (H2O2), and the activity of nitric oxide synthase (iNOS) were increased in excessive iron-fed sheep. Moreover, the present results revealed that excess iron was associated with a significant decrease in the activities of antioxidant capacity, such as glutathione peroxidase (GPx) activity and total antioxidant capacity (T-AOC) levels. The iNOS mRNA expression declined in excessive iron-fed sheep, indicating that down-regulation is likely to occur at the transcription level, which is consistent with the studies of iron blockades iNOS transcription. Surprisingly, the activity of glutathione peroxidase 1 (GPx1) continued to decline, but the expression levels of GPX1 mRNA and protein increased first and then decreased. This suggests that at the transcriptional and translation levels, the body compensatively increases the amount of GPx1 to maintain the balance of the oxidation-antioxidant system to resist peroxidation. Hematoxylin-eosin (HE) staining revealed histopathological changes in immune organs, such as lymphocyte infiltration and cell death, indicating that excessive iron-induced oxidative damage indirectly affects the body's immune function. These findings confirm the role of iron in regulating the homeostasis of the oxidation-antioxidant system.