Figure 4

Iron-induced pro-oxidant effects in human and murine cardiomyocytes and in murine models of iron-overload are prevented by RSV.

(A,B) Isolated adult LV human cardiomyocytes display a pronounced pro-oxidant phenotype after exposure to iron with increased dihydroethidium (DHE) staining for superoxide levels (top), 4-hydroxynonenal (4-HNE) immunofluorescence (middle), nitrotyrosine (NT) immunofluorescence (bottom) (A) and quantification of oxidative stress (B), while resveratrol (RSV; 100 μM) markedly suppressed iron-induced cellular oxidative stress. (C,D) Murine LV cardiomyocytes mirrored similar responses to iron as seen in human LV cardiomyocytes and iron-mediated cellular oxidative stress as illustrated by increased DHE staining, 4-HNE and nitrotyrosine immunofluorescence (C) and quantification of oxidative stress (D) was markedly suppressed by treatment with RSV. DHE fluorescence and is predominantly nuclear while 4-HNE and nitrotyrosine immunofluorescence are more diffuse and highlighted by the white arrows. n = 4 for immunofluorescence analysis; n = 8 for biochemical and gene expression analysis. *p < 0.05 compared with all other groups; ^#p < 0.05 compared with the placebo group.