LeBel C P, Ischiropoulos H, Bondy S C
Alkermes, Inc., Cambridge, Massachusetts 02139.
Chem Res Toxicol. 1992 Mar-Apr;5(2):227-31. doi: 10.1021/tx00026a012.
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-Fe(2+)-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.
在水介质中研究了使用二氯荧光素（DCFH）作为活性氧物质的一种测量方法。过氧化氢将DCFH氧化为荧光性的二氯荧光素（DCF），并且通过添加亚铁离子来放大这种氧化作用。在亚铁离子存在下，过氧化氢诱导的DCF形成被去铁胺完全抑制，被乙二胺四乙酸部分抑制，但被二乙烯三胺五乙酸增强。铁 - 过氧化物诱导的DCFH氧化被过氧化氢酶部分抑制，但不被辣根过氧化物酶抑制。DCFH的非螯合铁 - 过氧化物氧化被几种羟基自由基清除剂部分抑制，但与清除剂浓度无关，这表明在该系统中游离羟基自由基不参与DCFH的氧化。超氧阴离子不会直接氧化DCFH。数据表明，H2O2 - Fe(2+)衍生的氧化剂主要负责DCFH的非酶促氧化。此外，单独的过氧化物酶以及过氧化物酶还原H2O2过程中形成的氧化剂会氧化DCFH。由于DCFH氧化可能源自几种反应性中间体，因此在解释生物系统中涉及的特定活性氧物质时应谨慎。然而，DCFH作为毒理学现象中氧化应激的总体指标仍然是一种有吸引力的探针。
