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通过彗星试验测定的甲酰胺嘧啶DNA糖基化酶(Fpg)敏感位点、长期接触稀释剂吸入过程中丙二醛增加和谷胱甘肽减少之间的相关性。

Correlation between formamidopyrimidine DNA glycosylase (Fpg)-sensitive sites determined by a comet assay, increased MDA, and decreased glutathione during long exposure to thinner inhalation.

作者信息

Martínez-Alfaro M, Palma-Tirado L, Sandoval-Zapata F, Cárabez-Trejo A

机构信息

Departamento de Neurobiología del Desarrollo y Neurofisiología, Instituto de Neurobiología, Campus UNAM-Juriquilla, Apdo, Postal 1-1141, Querétaro 76001, México.

出版信息

Toxicol Lett. 2006 Jun 1;163(3):198-205. doi: 10.1016/j.toxlet.2005.10.021.

Abstract

Thinner inhalation causes toxic effects in a variety of organs, principally in the central nervous system. Some studies have shown oxidative stress effects of thinner inhalation, such as: activation of free radical processes, decrease of antioxidants, and oxidation products of proteins and lipids but not of DNA. The aim of this study is to investigate the effect of thinner inhalation on DNA. We used the comet assay in conjunction with the enzyme formamidopyrimidine glycoslyase (Fpg). Our results show a significant increase in Fpg-sensitive sites in DNA of lymphocytes from rats exposed to thinner fumes compared to lymphocytes from control rats (p < 0.05). Moreover, DNA damage detected with Fpg shows a high correlation with increased malondialdehyde (MDA) and decreased glutathione (GSH), two widely used biomarkers of oxidative stress. The most abundant base oxidation product found in DNA is 8-oxoguanine; it is the main substrate of Fpg and the most commonly used biomarker for oxidative DNA damage. This suggests that oxidative DNA damage is at least partly responsible for the DNA damage detected by Fpg. We propose the comet assay in combination with Fpg as a sensitive biomarker to monitor exposure to thinner inhalation. Limitations of this method are discussed.

摘要

吸入稀释剂会对多种器官产生毒性作用,主要是对中枢神经系统。一些研究表明吸入稀释剂会产生氧化应激效应,例如:自由基过程的激活、抗氧化剂的减少以及蛋白质和脂质而非DNA的氧化产物。本研究的目的是调查吸入稀释剂对DNA的影响。我们将彗星试验与酶甲酰胺嘧啶糖基化酶(Fpg)结合使用。我们的结果显示,与对照大鼠的淋巴细胞相比,暴露于稀释剂烟雾的大鼠淋巴细胞DNA中Fpg敏感位点显著增加(p < 0.05)。此外,用Fpg检测到的DNA损伤与氧化应激的两种广泛使用的生物标志物丙二醛(MDA)增加和谷胱甘肽(GSH)减少高度相关。在DNA中发现的最丰富的碱基氧化产物是8-氧代鸟嘌呤;它是Fpg的主要底物,也是氧化DNA损伤最常用的生物标志物。这表明氧化DNA损伤至少部分是Fpg检测到的DNA损伤的原因。我们提出将彗星试验与Fpg结合作为监测吸入稀释剂暴露的敏感生物标志物。讨论了该方法的局限性。

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