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首页> 美国卫生研究院文献>Toxicological Research >Evaluation of Oxidative DNA Damage Using an Alkaline Single Cell Gel Electrophoresis (SCGE) Comet Assay and the Protective Effects of N-Acetylcysteine Amide on Zearalenone-induced Cytotoxicity in Chang Liver Cells
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Evaluation of Oxidative DNA Damage Using an Alkaline Single Cell Gel Electrophoresis (SCGE) Comet Assay and the Protective Effects of N-Acetylcysteine Amide on Zearalenone-induced Cytotoxicity in Chang Liver Cells

机译：碱性单细胞凝胶电泳（SCGE）彗星试验评估氧化性DNA损伤以及N-乙酰半胱氨酸酰胺对玉米赤霉烯酮诱导的常肝细胞毒性的保护作用

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Zearalenone (ZEN) is a non-steroidal estrogenic mycotoxin produced by several species of Fusarium that are found in cereals and agricultural products. ZEN has been implicated in mycotoxicosis in farm animals and in humans. The toxic effects of ZEN are well known, but the ability of an alkaline Comet assay to assess ZEN-induced oxidative DNA damage in Chang liver cells has not been established. The first aim of this study was to evaluate the Comet assay for the determination of cytotoxicity and extent of DNA damage induced by ZEN toxin, and the second aim was to investigate the ability of N-acetylcysteine amide (NACA) to protect cells from ZEN-induced toxicity. In the Comet assay, DNA damage was assessed by quantifying the tail extent moment (TEM; arbitrary unit) and tail length (TL; arbitrary unit), which are used as indicators of DNA strand breaks in SCGE. The cytotoxic effects of ZEN in Chang liver cells were mediated by inhibition of cell proliferation and induction of oxidative DNA damage. Increasing the concentration of ZEN increased the extent of DNA damage. The extent of DNA migration, and percentage of cells with tails were significantly increased in a concentration-dependent manner following treatment with ZEN toxin (p < 0.05). Treatment with a low concentration of ZEN toxin (25 μM) induced a relatively low level of DNA damage, compared to treatment of cells with a high concentration of ZEN toxin (250 μM). Oxidative DNA damage appeared to be a key determinant of ZEN-induced toxicity in Chang liver cells. Significant reductions in cytolethality and oxidative DNA damage were observed when cells were pretreated with NACA prior to exposure to any concentration of ZEN. Our data suggest that ZEN induces DNA damage in Chang liver cells, and that the antioxidant activity of NACA may contribute to the reduction of ZEN-induced DNA damage and cytotoxicity via elimination of oxidative stress.
机译：玉米赤霉烯酮（ZEN）是一种非甾体类雌激素真菌毒素，由谷物和农产品中的几种镰刀菌产生。 ZEN与农场动物和人类的霉菌毒素中毒有关。 ZEN的毒性作用是众所周知的，但是碱彗星试验无法评估ZEN诱导的Chang肝细胞氧化DNA损伤的能力。这项研究的第一个目标是评估彗星测定法，以确定ZEN毒素诱导的细胞毒性和DNA损伤程度，第二个目标是研究N-乙酰半胱氨酸酰胺（NACA）保护细胞免受ZEN-诱导毒性。在彗星试验中，通过量化尾巴延伸矩（TEM；任意单位）和尾巴长度（TL；任意单位）来评估DNA损伤，这些尾随矩被用作SCGE中DNA链断裂的指标。 ZEN在长肝细胞中的细胞毒性作用是通过抑制细胞增殖和诱导氧化性DNA损伤来介导的。 ZEN浓度的增加会增加DNA损伤的程度。用ZEN毒素处理后，DNA迁移的程度和带有尾巴的细胞百分比显着增加（p <0.05）。与高浓度ZEN毒素（250μM）的细胞处理相比，低浓度ZEN毒素（25μM）的处理诱导了相对较低水平的DNA损伤。氧化DNA损伤似乎是ZEN诱导的Chang肝细胞毒性的关键决定因素。当细胞在暴露于任何浓度的ZEN之前用NACA预处理时，观察到细胞致死率和DNA氧化损伤的显着降低。我们的数据表明ZEN诱导了Chang肝细胞中的DNA损伤，而NACA的抗氧化活性可能通过消除氧化应激来降低ZEN诱导的DNA损伤和细胞毒性。

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