Long-term aerobic exercise increases redox-active iron through nitric oxide in rat hippocampus

Abstract

Adult hippocampus is highly vulnerable to iron-induced oxidative stress. Aerobic exercise has been proposed to reduce oxidative stress but the findings in the hippocampus are conflicting. This study aimed to observe the changes of redox-active iron and concomitant regulation of cellular iron homeostasis in the hippocampus by aerobic exercise, and possible regulatory effect of nitric oxide (NO). A randomized controlled study was designed in the rats with swimming exercise treatment (for 3 months) and/or an unselective inhibitor of NO synthase (NOS) (L-NAME) treatment. The results from the bleomycin-detectable iron assay showed additional redox-active iron in the hippocampus by exercise treatment. The results from nonheme iron content assay, combined with the redox-active iron content, showed increased storage iron content by exercise treatment. NOx (nitrate plus nitrite) assay showed increased NOx content by exercise treatment. The results from the Western blot assay showed decreased ferroportin expression, no changes of TfR1 and DMT1 expressions, increased IRP1 and IRP2 expression, increased expressions of eNOS and nNOS rather than iNOS. In these effects of exercise treatment, the increased redox-active iron content, storage iron content, IRP1 and IRP2 expressions were completely reversed by L-NAME treatment, and decreased ferroportin expression was in part reversed by L-NAME. L-NAME treatment completely inhibited increased NOx and both eNOS and nNOS expression in the hippocampus. Our findings suggest that aerobic exercise could increase the redox-active iron in the hippocampus, indicating an increase in the capacity to generate hydroxyl radicals through the Fenton reactions, and aerobic exercise-induced iron accumulation in the hippocampus might mainly result from the role of the endogenous NO.

Keywords: AE; ANOVA; BDI; DMT1+IRE; DMT1−IRE; DTPA; E1; E2; Exercise; Fpn; HC; Hippocampus; IRE; IRP1; IRP2; Iron metabolism; L-NAME; MDA; N(ω)-nitro-L-arginine methyl ester; NHI; NO; NOS; NOx; Nitric oxide; ROS; Redox-active iron; S1; S2; SEM; Storage iron; TCA; TfR1; aerobic exercise; bleomycin-detectable iron; diethylene triamine pentacetate acid; divalent metal transporter 1 with iron responsive element; divalent metal transporter 1 without iron responsive element; eNOS; endothelial nitric oxide synthase; exercise group; exercise+L-NAME group; ferroportin 1; hippocampus; iNOS; inducible nitric oxide synthase; iron regulatory protein 1; iron regulatory protein 2; iron responsive element; malondialdehyde; nNOS; neuronal nitric oxide synthase; nitric oxide; nitric oxide synthase; nitrite plus nitrate; nonheme iron; reactive oxygen species; sedentary group; sedentary+L-NAME group; standard error of mean; transferrin receptor 1; trichloroacetic acid; two-way analysis of variance.