Sudan I induces genotoxic effects and oxidative DNA damage in HepG2 cells
Sudan I induces genotoxic effects and oxidative DNA damage in HepG2 cells
复制标题
DOI：
10.1016/j.mrgentox.2006.11.004
复制
发表时间：
2007-03-05
影响因子：
1.9
通讯作者：
Zhong, Laifu
中科院分区：
文献类型：
--
作者：
An, Yu;Jiang, Liping;Zhong, Laifu
苏丹1是一种合成脂溶性偶氮颜料，广泛应用于各个工业领域。然而，苏丹一号尚未在任何级别的食品生产中获得批准，因为有许多关于其对人类的遗传毒性和致癌性的不确定报告。本研究的目的是评估苏丹 I 的基因毒性作用，并通过使用人肝癌 HepG2 细胞来鉴定和阐明反应机制。为了研究苏丹1号的基因毒性作用，使用了彗星试验和微核试验(MNT)。在彗星试验和 MNT 中，我们发现苏丹 I 在所有测试浓度（25-100 μM）下 DNA 迁移和微核频率均以剂量依赖性方式增加。数据表明苏丹一号导致DNA链断裂和染色体断裂。为了阐明这种差异的根本机制，我们使用 2,7-二氯荧光素二乙酸酯测定法监测了活性氧 (ROS) 的产生水平。使用 8-羟基脱氧鸟苷 (8-OHdG) 的免疫过氧化物酶染色并通过测量硫代巴比妥酸反应物质 (TBARS) 的水平来评估氧化 DNA 损伤和脂质过氧化的水平。在较高浓度下，剂量分别为100、50-100和50-100μM，在HepG2细胞中观察到ROS、8-OHdG和TBARS水平显着增加。我们得出结论，苏丹 I 可能通过较高剂量下 ROS 诱导的氧化 DNA 损伤而引起基因毒性作用。 (c) 2006 Elsevier B.V. 保留所有权利。
Sudan 1, a synthetic lipid soluble azo pigment, is widely used in various industrial fields. However, Sudan I has not been approved at any level of food production, since there are many inconclusive reports relating to its genotoxicity and carcinogenicity in humans. The aim of this study was to assess the genotoxic effects of Sudan I and to identifiy and clarify the reaction mechanisms by use of human hepatoma HepG2 cells. To study the genotoxic effects of Sudan 1, the comet assay and micronucleus test (MNT) were used. In the comet assay and MNT, we found increase of DNA migration and of the micronuclei frequencies at all tested concentrations (25-100 mu M) of Sudan I in a dose-dependent manner. The data suggest that Sudan I caused DNA strand breaks and chromosome breaks. To elucidate the underlying mechanism of this difference, we monitored the level of reactive oxygen species (ROS) production with the 2,7-dichlorofluorescein diacetate assay. The level of the oxidative DNA damage and lipid peroxidation was evaluated using immunoperoxidase staining for 8-hydroxydeoxyguanosine (8-OHdG) and by measuring levels of thiobarbituric acid-reactive substances (TBARS). Significantly increased levels of ROS, 8-OHdG and TBARS were observed in HepG2 cells at higher concentrations, the doses being 100, 50-100 and 50-100 mu M, respectively. We conclude that Sudan I causes genotoxic effects, probably via ROS-induced oxidative DNA damage at the higher doses. (c) 2006 Elsevier B.V. All rights reserved.