EVALUATION OF THE PROBE 2',7'-DICHLOROFLUORESCIN AS AN INDICATOR OF REACTIVE OXYGEN SPECIES FORMATION AND OXIDATIVE STRESS
EVALUATION OF THE PROBE 2',7'-DICHLOROFLUORESCIN AS AN INDICATOR OF REACTIVE OXYGEN SPECIES FORMATION AND OXIDATIVE STRESS
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DOI：
10.1021/tx00026a012
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发表时间：
1992-03-01
影响因子：
4.1
通讯作者：
BONDY, SC
中科院分区：
文献类型：
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作者：
LEBEL, CP;ISCHIROPOULOS, H;BONDY, SC
研究了在水介质中用二氯荧光素(DCFH)作为测定活性氧物种的方法。过氧化氢将DCFH氧化成荧光二氯荧光素(DCF)，加入亚铁后，氧化作用被放大。亚铁离子存在下过氧化氢诱导的DCF形成可被去铁胺完全抑制，可被乙二胺四乙酸部分抑制，但可被二乙三胺五乙酸增强。过氧化氢酶可部分抑制过氧化铁诱导的DCFH氧化，而辣根过氧化物酶不能抑制过氧化铁诱导的DCFH氧化。DCFH的非螯合过氧化铁氧化可被几种羟基自由基清除剂部分抑制，但不受清除剂浓度的影响，这表明自由基不参与DCFH的氧化。超氧阴离子不直接氧化DCFH。数据表明，双氧水-Fe2+衍生的氧化剂是DCFH非酶氧化的主要原因。此外，过氧化物酶单独作用和过氧化物酶还原过氧化氢过程中形成的氧化剂可氧化DCFH。由于DCFH氧化可能来自几个活性中间体，因此对生物系统中涉及的特定活性氧物种的解释应谨慎处理。然而，DCFH仍然是一个有吸引力的指标，作为毒理学现象中氧化应激的总体指标。
The use of dichlorofluorescin (DCFH) as a measure of reactive oxygen species was studied in aqueous media. Hydrogen peroxide oxidized DCFH to fluorescent dichlorofluorescein (DCF), and the oxidation was amplified by the addition of ferrous iron. Hydrogen peroxide-induced DCF formation in the presence of ferrous iron was completely inhibited by deferoxamine and partially inhibited by ethylenediaminetetraacetic acid, but was augmented by diethylenetriaminepentaacetic acid. Iron-peroxide-induced oxidation of DCFH was partially inhibited by catalase but not by horseradish peroxidase. Nonchelated iron-peroxide oxidation of DCFH was partially inhibited by several hydroxyl radical scavengers, but was independent of the scavenger concentration, and this suggests that free hydroxyl radical is not involved in the oxidation of DCFH in this system. Superoxide anion did not directly oxidize DCFH. Data suggest that H2O2-Fe2+-derived oxidant is mainly responsible for the nonenzymatic oxidation of DCFH. In addition, peroxidase alone and oxidants formed during the reduction of H2O2 by peroxidase oxidize DCFH. Since DCFH oxidation may be derived from several reactive intermediates, interpretation of specific reactive oxygen species involved in biological systems should be approached with caution. However, DCFH remains an attractive probe as an overall index of oxidative stress in toxicological phenomena.