title: In vitro interceptive and reparative effects of myo-inositol against copper-induced oxidative damage and antioxidant system disturbance in primary cultured fish enterocytes.
abstract: Copper (Cu) is essential for normal cellular processes in most eukaryotic organisms but is toxic in excess. Our previous study reported that a nutrient antioxidant, myo-inositol (MI), can protect fish from Cu-induced oxidative injury; however, the mechanisms involved are not fully understood. Therefore, the present study aimed to analyze potential pathways. First, to investigate the hypothesis that MI protects enterocytes against Cu toxicity via the intercept pathway, enterocytes were treated with different concentrations of MI (0-75mg/L medium) in the presence of 6mg/L of Cu for 24h (Experiment 1). Next, we investigated the potential reparative role of MI after a Cu challenge (Experiment 2). The results of Experiment 1 indicated that cells exposed to Cu alone for 24h exhibited increases in lactate dehydrogenase release (LDH), malondialdehyde (MDA) formation and protein oxidation (P<0.05). Notably, a dose-dependent inhibitory effect on LDH release was observed with all doses of MI. Moreover, co-treatment with MI completely inhibited Cu-induced protein carbonyl (PC) formation. However, Cu-induced lipid peroxidation was not altered by MI co-treatment. Additionally, Cu exposure suppressed total-superoxide dismutase (T-SOD), CuZnSOD and catalase (CAT) activities, and these changes were completely blocked by co-treatment with sufficient MI concentrations. In contrast, cells exposed to Cu exhibited adaptive increases in reduced glutathione (GSH) content and the activities of anti-hydroxyl radical (AHR), glutathione peroxidase (GPx), glutathione-S-transferase (GST) and glutathione reductase (GR). Interestingly, the Cu-stimulated increases in these antioxidants were blocked by co-treatment with sufficient MI concentrations. The results of Experiment 2 indicated that cell injury (LDH release), lipid peroxidation (MDA formation) and protein oxidation induced by Cu were reversed by subsequent MI treatment. Meanwhile, Cu-induced decreases in alkaline phosphatase (AKP), anti-superoxide anion (ASA), T-SOD and CuZnSOD activities were completely restored by subsequent MI treatment, while the reduced CAT activity was partially restored. However, MI rescues partially restored the adaptive increase in GPx activity induced by Cu, whereas the adaptive increase in reduced GSH content was completely reversed by 75mg/L of MI. However, subsequent MI treatments did not alter the induction of GST activity by Cu. In conclusion, we demonstrated for the first time that MI not only protected enterocytes from Cu-induced oxidative damage but also increased the repair activity in primary enterocytes after challenge with Cu. Moreover, MI-mediated increases in antioxidant enzyme activities contributed to lipid and protein oxidant repair.
authors.affiliation: Animal Nutrition Institute, Sichuan Agricultural University, Ya'an 625014, Sichuan, China.
authors.forename: Wei-Dan
authors.initials: WD
authors.lastname: Jiang
authors.forename: Yang
authors.initials: Y
authors.lastname: Liu
authors.forename: Jun
authors.initials: J
authors.lastname: Jiang
authors.forename: Kai
authors.initials: K
authors.lastname: Hu
authors.forename: Shu-Hong
authors.initials: SH
authors.lastname: Li
authors.forename: Lin
authors.initials: L
authors.lastname: Feng
authors.forename: Xiao-Qiu
authors.initials: XQ
authors.lastname: Zhou
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000378:metabolism*; D002347:Carps / Q000378:metabolism*; D003300:Copper / Q000633:toxicity*; D020895:Enterocytes / Q000187:drug effects / Q000378:metabolism; D007294:Inositol / Q000008:administration & dosage / Q000494:pharmacology*; D007413:Intestinal Mucosa / Q000378:metabolism; D007422:Intestines / Q000187:drug effects; D010084:Oxidation-Reduction; D018384:Oxidative Stress / Q000187:drug effects*; D014874:Water Pollutants, Chemical / Q000633:toxicity*
chemical_list: D000975:Antioxidants; D014874:Water Pollutants, Chemical; D007294:Inositol; D003300:Copper