α,β-unsaturated carbonyl compounds:: induction of oxidative DNA damage in mammalian cells
α,β-unsaturated carbonyl compounds:: induction of oxidative DNA damage in mammalian cells
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DOI：
10.1093/mutage/geg018
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发表时间：
2003-09-01
期刊：
影响因子：
2.7
通讯作者：
Eisenbrand, G
中科院分区：
文献类型：
--
作者：
Janzowski, C;Glaab, V;Eisenbrand, G
α，β-不饱和羰基化合物存在于食品和其他环境介质中。由于它们与细胞亲核试剂的反应性（例如与DNA碱基和谷胱甘肽形成迈克尔加合物），它们可能代表潜在的健康风险。在这项研究中，在哺乳动物细胞中研究了氧化性DNA损伤的诱导，这是由选定的食物相关的2-烯醛（包括E-（2）-己烯醛（HEX）、（2 E，4 E）-2，4-己二烯醛（HEXDI）和（E）-2-肉桂醛（CA））以及环状类似物2-环己烯-1-酮（CHX）诱导的谷胱甘肽耗竭的结果。用彗星试验监测氧化性DNA断裂，使用甲酰胺基嘧啶-DNA糖基化酶（FPG）处理。通过动力学光度测定法测定总细胞谷胱甘肽（tGSH）。V79细胞与HEX（100 μ M）和CHX（300 μ M）、HEXDI和CA（各300 μ M）孵育1小时后，tGSH被耗尽至85%。在这些条件下，未观察到FPG敏感位点;然而，可检测到中度直接DNA断裂。在孵育后3 h期间（无供试化合物），HEX-和CA-预处理的细胞中发生了明显的氧化性DNA断裂，但CHX-和HEXDI-预处理的细胞中未发生氧化性DNA断裂。直接DNA断裂显着减少，最有可能的修复过程中，和tGSH浓度观察到治疗后3小时内再次增加。这些结果为烯醇介导的氧化应激导致细胞毒性/遗传毒性细胞损伤提供了强有力的证据。氧化应激的程度似乎受到烯醛的结构特异性的影响。
alpha,beta-Unsaturated carbonyl compounds occur in food and other environmental media. Due to their reactivity with cellular nucleophiles (e.g. Michael adduct formation with DNA bases and with glutathione) they might represent a potential health risk. In this study, induction of oxidative DNA damage was investigated in mammalian cells, as a consequence of glutathione depletion induced by selected food relevant 2-alkenals, including E-(2)-hexenal (HEX), (2E,4E)-2,4-hexadienal (HEXDI) and (E)-2-cinnamaldehyde (CA) and the cyclic analogue 2-cyclohexen-1-one (CHX). Oxidative DNA breakage was monitored with the Comet assay, using treatment with formamidopyrimidine-DNA glycosylase (FPG). Total cellular glutathione (tGSH) was determined in a kinetic, photometric assay. After 1 h incubation of V79 cells with HEX (100 muM) and CHX (300 muM), HEXDI and CA (300 muM each), tGSH was depleted down to 85%). Under these conditions, FPG-sensitive sites were not observed; moderate direct DNA breakage, however, was detectable. During 3 h post-incubation (without test compound) distinct oxidative DNA breakage occurred in HEX- and CA-, but not in CHX- and HEXDI-pretreated cells. Direct DNA breakage was markedly diminished, most probably by repair processes, and tGSH concentrations were observed to increase again within 3 h post-treatment. The results give strong evidence for alkenal-mediated oxidative stress contributing to cytotoxic/genotoxic cell damage. The extent of oxidative stress appears to be influenced by structure-specific properties of the alkenals.