Brömme Hans-Jürgen, Zühlke Leoni, Silber Rolf-Edgar, Simm Andreas
Institut für Pathophysiologie, Martin-Luther-Universität Halle-Wittenberg, Ernst-Grube-Str. 40, D-06097 Halle, Germany.
Klinik für Herz- und Thoraxchirurgie, Martin-Luther-Universität Halle-Wittenberg, Ernst-Grube-Str. 40, D-06097 Halle, Germany.
Exp Gerontol. 2008 Jul;43(7):638-644. doi: 10.1016/j.exger.2008.01.010. Epub 2008 Feb 2.
The oxidation of 2',7'-dichlorofluorescin (DCFH(2)) is widely used as a measure to detect the generation of reactive oxygen species (ROS) and to analyze oxidative stress. Other factors beside commonly known radicals may influence the results of such measurements. Therefore, the effects of H(2)O(2), KMnO(4,) decomposition products of AAPH, ethanol and DMSO, antioxidants like ascorbic acid, different ferrous ion chelates, and heme-containing proteins like cytochrome c, myoglobin, hemoglobin, and horseradish-peroxidase were comparatively analyzed with respect to their impact on DCFH(2) oxidation. The study evaluates the effects of various oxidants with different oxidative potentials regarding their ability to induce DCF-fluorescence. Furthermore, we analyzed the inhibitory effect of organic solvents like ethanol or DMSO on the oxidation of DCFH(2) by hydroxyl radicals. The results of our study indicate that the potential of an oxidant does not always correlate with its efficiency to generate DCF-fluorescence.
2',7'-二氯荧光素(DCFH(2))的氧化被广泛用作检测活性氧(ROS)生成和分析氧化应激的一种手段。除了常见的自由基外,其他因素也可能影响此类测量结果。因此,我们比较分析了过氧化氢(H(2)O(2))、高锰酸钾(KMnO(4))、偶氮二异丁腈(AAPH)分解产物、乙醇和二甲基亚砜(DMSO)、抗氧化剂如抗坏血酸、不同的亚铁离子螯合物以及含血红素的蛋白质如细胞色素c、肌红蛋白、血红蛋白和辣根过氧化物酶对DCFH(2)氧化的影响。该研究评估了具有不同氧化电位的各种氧化剂诱导DCF荧光的能力。此外,我们分析了乙醇或DMSO等有机溶剂对羟基自由基氧化DCFH(2)的抑制作用。我们的研究结果表明,氧化剂的电位并不总是与其产生DCF荧光的效率相关。