Peroxide-triggered erythrocytes haemolysis as a model for the study of oxidative damage in marine fishes

The haemolysis of sea bass Dicentrarchus labrax red blood cells (RBC) was initiated by tert -butyl-hydroperoxide (t-BHP). The onset of the haemolytic process was accelerated by increasing t-BHP concentration. This process was preceded by a drop in the RBC glutathione content followed by the production of lipid peroxidation products. Also t-BHP induced DNA fragmentation in RBC nuclei as measured by COMET assay. The addition of the antioxidant Trolox C® dose-dependently delayed the onset of both lipid peroxidation and haemolysis, and protected GSH stores against t-BHP-induced depletion. DNA fragmentation was also pre-vented by Trolox C®. These results indicate that t-BHP induces haemolysis in sea bass RBC through the induction of oxidative stress. Such a simple model could prove useful for both fundamental and applied studies on marine fish antioxidant mechanisms.