超氧自由基阴离子和过氧化氢检测的最新进展：氧化还原信号传导的机遇、挑战和影响。,Archives of Biochemistry and Biophysics

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超氧自由基阴离子和过氧化氢检测的最新进展：氧化还原信号传导的机遇、挑战和影响。
Archives of Biochemistry and Biophysics ( IF 3.8 ) Pub Date : 2016-08-30 , DOI: 10.1016/j.abb.2016.08.021
Balaraman Kalyanaraman ₁ , Micael Hardy ₂ , Radoslaw Podsiadly ₃ , Gang Cheng ₁ , Jacek Zielonka ₁

Affiliation

在这篇综述中，讨论了针对超氧化物 (O2-) 和过氧化氢 (H2O2) 的探针和检测技术的一些最新进展。在过去的十年中，由于新的氧化还原探针的合成、对其化学性质的更好理解以及特异性和灵敏检测方法的开发，O2 和 H2O2 检测取得了重大进展。对于超氧化物检测，氢乙啶 (HE) 是最合适的探针，因为产品 2-羟基乙锭对 O2- 具有特异性。此外，HE 衍生的二聚体产物对单电子氧化剂具有特异性。由于红色荧光乙锭总是由 HE 在细胞内形成，因此需要色谱技术来检测 2-羟基乙锭。HE 类似物 Mito-SOX 和氢丙啶与 O2- 和单电子氧化剂表现出相同的反应化学。因此，可以使用基于 HPLC 的方法而不是荧光显微镜明确检测线粒体超氧化物。芳香族硼酸盐探针与 H2O2 发生定量反应，形成酚类产物。然而，过氧亚硝酸盐和次氯酸盐与硼酸盐反应更快，形成相同的产物。使用 ROS 特异性探针和 HPLC 检测，可以筛选化学库以发现 NADPH 氧化酶的特异性抑制剂。我们希望对不同细胞区室中的 O2- 和 H2O2 进行严格检测将增进我们对它们在氧化还原信号传导中的作用的理解。

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Recent developments in detection of superoxide radical anion and hydrogen peroxide: Opportunities, challenges, and implications in redox signaling.

In this review, some of the recent developments in probes and assay techniques specific for superoxide (O2-) and hydrogen peroxide (H2O2) are discussed. Over the last decade, significant progress has been made in O2- and H2O2 detection due to syntheses of new redox probes, better understanding of their chemistry, and development of specific and sensitive assays. For superoxide detection, hydroethidine (HE) is the most suitable probe, as the product, 2-hydroxyethidium, is specific for O2-. In addition, HE-derived dimeric products are specific for one-electron oxidants. As red-fluorescent ethidium is always formed from HE intracellularly, chromatographic techniques are required for detecting 2-hydroxyethidium. HE analogs, Mito-SOX and hydropropidine, exhibit the same reaction chemistry with O2- and one-electron oxidants. Thus, mitochondrial superoxide can be unequivocally detected using HPLC-based methods and not by fluorescence microscopy. Aromatic boronate-based probes react quantitatively with H2O2, forming a phenolic product. However, peroxynitrite and hypochlorite react more rapidly with boronates, forming the same product. Using ROS-specific probes and HPLC assays, it is possible to screen chemical libraries to discover specific inhibitors of NADPH oxidases. We hope that rigorous detection of O2- and H2O2 in different cellular compartments will improve our understanding of their role in redox signaling.

更新日期：2016-08-30

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