Abstract
Although oxidative stress has been demonstrated to be involved in copper (Cu)-induced toxicity, information regarding the effect of antioxidants on Cu toxicity is still scarce. This study assessed the possible protective effects of myo-inositol (MI) against subsequent Cu exposure in juvenile Jian carp ( Cyprinus carpio var. Jian) in vivo and in their enterocytes in vitro. First, oxidative stress was established by exposing fish to different concentrations of Cu (0–7.2 mg Cu/L water) for 4 days. Next, the protective effects of MI (administered as a dietary supplement for 60 days) against subsequent Cu exposure (0.6 mg Cu/L water for 4 days) were studied in fish. The third trial determined the effects of Cu exposure (0–6.0 mg Cu/L of medium for 24 h) on enterocytes in vitro. Finally, enterocytes were pre-incubated with graded levels of MI (0–75 mg MI/L of medium) for 72 h and exposed to 6.0 mg Cu/L of medium for 24 h. The results indicated that ≥0.6 mg Cu/L water could induce oxidative stress in fish ( P < 0.05). Cu exposure significantly induced increases in lipid peroxidation and protein oxidation in the gill, hepatopancreas and intestine in fish. However, these oxidative effects were prevented by MI pre-supplementation. MI also prevented the toxic effects of Cu on anti-superoxide anion (ASA), anti-hydroxyl radical (AHR), superoxide dismutase (SOD), catalase (CAT), glutathione- S-transferase (GST), glutathione peroxidase (GPx) and glutathione reductase (GR) activities and glutathione (GSH) content in these organs. In vitro, enterocytes exposed to Cu displayed a dose-dependent injury. Moreover, cell viability, protein retention (PR), alkaline phosphatase, total-SOD (T-SOD) and Cu/ZnSOD activities were all depressed by Cu ( P < 0.05). Interestingly, the final experiment showed that MI pre-supplementation could block the toxic effects of Cu on the antioxidant system, and thus protect enterocytes from Cu-induced oxidative damage. All of these results indicated that the induction of key antioxidant defenses by MI pre-supplementation, including SOD, CAT, GPx, GST and GSH, may play an important role in the protection of fish against oxidative stress.
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