In vitro interceptive and reparative effects of myo-inositol against copper-induced oxidative damage and antioxidant system disturbance in primary cultured fish enterocytes
In vitro interceptive and reparative effects of myo-inositol against copper-induced oxidative damage and antioxidant system disturbance in primary cultured fish enterocytes
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肌醇对原代养殖鱼肠细胞中铜诱导的氧化损伤和抗氧化系统干扰的体外拦截和修复作用。
DOI：
10.1016/j.aquatox.2013.02.005
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发表时间：
2013-05-15
影响因子：
4.5
通讯作者：
Zhou, Xiao-Qiu
中科院分区：
文献类型：
--
作者：
Jiang, Wei-Dan;Liu, Yang;Zhou, Xiao-Qiu
铜（Cu）是大多数真核生物正常细胞过程所必需的，但过量是有毒的。我们以前的研究报告，营养抗氧化剂，肌醇（MI），可以保护鱼类免受铜诱导的氧化损伤，然而，所涉及的机制并不完全清楚。因此，本研究旨在分析潜在的途径。首先，为了研究MI通过拦截途径保护肠细胞免受Cu毒性的假设，在6 mg/L Cu的存在下用不同浓度的MI（0-75 mg/L培养基）处理肠细胞24 h（实验1）。接下来，我们研究了铜挑战后MI的潜在修复作用（实验2）。实验1结果表明，单独铜暴露24 h，细胞乳酸脱氢酶（LDH）释放、丙二醛（MDA）生成和蛋白质氧化均显著增加（P < 0.05）。值得注意的是，所有剂量的MI均观察到对LDH释放的剂量依赖性抑制作用。此外，与MI共处理完全抑制铜诱导的蛋白质羰基（PC）的形成。然而，铜诱导的脂质过氧化反应没有改变MI共同治疗。此外，铜暴露抑制总超氧化物歧化酶（T-SOD），CuZnSOD和过氧化氢酶（CAT）的活性，这些变化被完全阻断了足够的MI浓度的共同处理。与此相反，铜暴露的细胞表现出适应性增加还原型谷胱甘肽（GSH）含量和抗羟基自由基（AHR），谷胱甘肽过氧化物酶（GPx），谷胱甘肽-S-转移酶（GST）和谷胱甘肽还原酶（GR）的活性。有趣的是，这些抗氧化剂的铜刺激的增加被阻断了足够的MI浓度的共处理。实验2的结果表明，随后的MI处理逆转了Cu诱导的细胞损伤（LDH释放），脂质过氧化（MDA形成）和蛋白质氧化。同时，铜诱导的碱性磷酸酶（AMP），抗超氧阴离子（阿萨），T-SOD和CuZnSOD活性的下降完全恢复随后的MI处理，而降低的CAT活性部分恢复。然而，MI救援部分恢复了铜诱导的GPx活性的适应性增加，而还原型GSH含量的适应性增加被75 mg/L的MI完全逆转。然而，随后的MI治疗没有改变诱导GST活性的铜。总之，我们第一次证明，MI不仅保护肠上皮细胞免受铜诱导的氧化损伤，但也增加了修复活动后，在原代肠上皮细胞与铜的挑战。此外，MI介导的抗氧化酶活性的增加有助于脂质和蛋白质氧化修复。(C)2013爱思唯尔有限公司版权所有。
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 (0-75 mg/L medium) in the presence of 6 mg/L of Cu for 24 h (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 24 h 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 (AMP), 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 75 mg/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. (C) 2013 Elsevier B.V. All rights reserved.