Interactions of free copper (II) ions alone or in complex with iron (III) ions with erythrocytes of marine fish Dicentrarchus labrax

As a consequence of human activity, various toxicants - especially metal ions - enter aquatic ecosystems and many fish are exposed to considerable levels. As the free ion and in some complexes, there is no doubt that copper promotes damage to cellular molecules and structures through radical formation. Therefore, we have investigated the influence of copper uptake by the red blood of the sea bass (Dicentrarchus labrax), and its oxidative action and effects on cells in the presence of complexed and uncomplexed Fe3+ ions. Erythrocytes were exposed to various concentrations of CuSO4, Fe(NO3)(3), and K3Fe(CN)(6) for up to 5 h, and the effects of copper ions alone and in the combination with iron determined. The results show that inside the cells cupric ion interacts with hemoglobin, causing methemoglobin formation by direct electron transfer from heme Fe2+ to Cu2+. Potassium ferricyanide as a source of complexed iron decreases Met-Hb formation induced by copper ions unlike Fe(NO3)(3). We also found that incubation of fish erythrocytes with copper increased hemolysis of cells. But complexed and uncomplexed iron protected the effect of copper. CuSO4 increased the level of lipid peroxidation and a protective effect on complexed iron was observed. Incubation of erythrocytes with copper ions resulted in the loss of a considerable part of thiol content at 10 and 20 mu M. This effect was decreased by potassium ferricyanide and Fe(NO3)(3) only after 1 and 3 h of incubation. The level of nuclear DNA damage assayed by comet assay showed that 20 mu M CuSO4 as well as 20 mu M Fe(NO3)(3) and 10 mM K3Fe(CN)(6) induce single- and double-strand breaks. The lower changes were observed after the exposure of cells to K3Fe(CN)(6). The data suggest that complexed iron can act protectively against copper ions in contrast to Fe(NO3)(3). (C) 2009 International Federation for Cell Biology. Published by Elsevier Ltd. All rights reserved.