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一种基于热释光的快速且易于操作的技术,用于评估四氯化碳对大鼠肝细胞诱导的氧化应激。

A rapid and easy to handle thermoluminescence based technique for evaluation of carbon tetrachloride-induced oxidative stress on rat hepatocytes.

作者信息

Schumann Anika, Bauer Alexander, Hermes Matthias, Gilbert Matthias, Hengstler Jan G, Wilhelm Christian

机构信息

Biology I, Plant Physiology, University of Leipzig, Johannisallee 21-23, 04103 Leipzig, Germany.

出版信息

Arch Toxicol. 2009 Jul;83(7):709-20. doi: 10.1007/s00204-009-0404-4. Epub 2009 Feb 12.

Abstract

Oxidative stress has become one of the most intensively studied topics in biomedical research and is an often observed mechanism of non-genotoxic carcinogens like carbon tetrachloride. To monitor the oxidative stress status in in vitro hepatocytes, we compared thermoluminescence (TL) measurements with biochemical standard methods for oxidative stress markers. In contrast to biochemical analysis, TL measurements can be performed without any time-consuming extraction procedures by using directly collected cell material. After incubation with CCl(4) (24 h), thermo-induced light emission increased with rising concentration of CCl(4) up to eightfold at 10 mM CCl(4). Simultaneously, we determined the content of different secondary oxidative stress products, like thiobarbituric acid reactive substances and malondialdehyde. The rise of all biochemical markers complied with the increasing concentration of CCl(4). Finally, we could show that the CCl(4)-induced increase of oxidative stress markers determined by time-consuming biochemical methods perfectly correlates with the increase of high temperature bands in rapid TL measurements.

摘要

氧化应激已成为生物医学研究中研究最为深入的课题之一,并且是诸如四氯化碳等非遗传毒性致癌物常见的作用机制。为监测体外培养肝细胞中的氧化应激状态,我们将热释光(TL)测量结果与用于氧化应激标志物的生化标准方法进行了比较。与生化分析不同,通过直接使用收集的细胞材料,热释光测量无需任何耗时的提取程序即可进行。用四氯化碳(CCl₄)孵育24小时后,热诱导发光随CCl₄浓度升高而增加,在10 mM CCl₄时增加至八倍。同时,我们测定了不同次级氧化应激产物的含量,如硫代巴比妥酸反应性物质和丙二醛。所有生化标志物的升高都与CCl₄浓度的增加一致。最后,我们可以证明,通过耗时的生化方法测定的CCl₄诱导的氧化应激标志物增加与快速热释光测量中高温带的增加完全相关。

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