Pyrene-stimulated reactive oxygen species generation and oxidative damage in Carassius auratus

Laboratory experiments were carried out to understand the toxicology of pyrene in the goldfish Carassius auratus and investigate the potential oxidative stress induced by reactive oxygen species (ROS) in vivo in a time-dependent manner. Pyrene bioaccumulation, induction of reactive oxygen species and the consequent biochemical responses in the liver of the fish were examined. Fish were exposed to 0.05 mg/L pyrene for different periods. The pyrene concentration in fish liver was analyzed by high performance liquid chromatography (HPLC). Free radicals were detected by electron paramagnetic resonance (EPR). The activities of antioxidant enzymes, contents of nonenzymatic antioxidants and malondialdehyde (MDA) in fish liver were also determined. Results indicated that the pyrene concentrations in fish liver reached a maximum level on day 1, and then declined to a low steady state level over 7 days. The free radical significantly increased at 6 h and reached a maximum on day 2, while the superoxide dismutase (SOD) activity and MDA content were induced, and the reduced glutathione (GSH) content was inhibited by day 2. The catalase (CAT) and glutathione-S-transferase (GST) activities were significantly induced at 12 h. These results indicated that pyrene was rapidly bioaccumulated in fish resulting in redox cycling, and the production of free radical is an important mechanism of pyrene toxicity in C. auratus. The indicators of antioxidant system are sensitive and useful for the study of early biomarkers of pyrene exposure in fish.