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Characterization of the Electrochemical Oxidation of Peroxynitrite: Relevance to Oxidative Stress Bursts Measured at the Single Cell Level

机译:过氧亚硝酸盐的电化学氧化的表征:与单细胞水平上测量的氧化应激爆发的相关性

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摘要

The electrochemical signature of peroxynitrite oxidation is reported for the first time, and its mechanism discussed in the light of data obtained by steady-state and transient voltammetry at microelectrodes. Peroxynitrite is an important biological species generated by aerobic cells presumably via the near diffusion-limited coupling of nitric oxide and superoxide ion. Its production by living cells has been previously suspected during cellular oxidative bursts as well as in several human pathologies (arthritis, inflammation, apoptosis, ageing, carcinogenesis, Alzheimer disease, AIDS, etc.). However, this could only be inferred on the basis of characteristic patient metabolites or through indirect detection, or by observation of follow-up species resulting supposedly from its chemical reactions in vivo. In this work, thanks to the independent knowledge of the electrochemical characteristics of ONO_2~- oxidation, the kinetics and intensity of this species released by single human fibroblasts could be established directly and quantitatively based on the application of the artificial synapse method. It was then observed and established that fibroblasts submitted to mechanical stresses produce oxidative bursts, which involve the release within less than a tenth of a second of a complex cocktail composed of several femtomoles of peroxynitrite, hydrogen peroxide, nitric oxide, and nitrite ions.
机译:首次报道了过氧亚硝酸盐氧化的电化学特征,并根据稳态和瞬时伏安法在微电极上获得的数据讨论了其机理。过氧亚硝酸盐是好氧细胞产生的重要生物物种,大概是通过一氧化氮和超氧离子的近扩散限制耦合而产生的。以前曾怀疑在细胞氧化爆发期间以及几种人类病理学(关节炎,炎症,细胞凋亡,衰老,致癌作用,阿尔茨海默氏病,艾滋病等)中,活细胞会产生它。但是,只能根据患者的特征代谢产物或通过间接检测或通过观察据认为是由其体内化学反应产生的后续物种来推断。在这项工作中,由于对ONO_2〜-氧化的电化学特性有独立的了解,因此可以基于人工突触方法的应用,直接和定量地确定单个人成纤维细胞释放的该物种的动力学和强度。然后观察到并确定,经受机械应力的成纤维细胞产生氧化爆发,该爆发涉及在不到十分之一秒的时间内释放出由过氧化亚硝酸盐,过氧化氢,一氧化氮和亚硝酸根离子的几个飞摩尔组成的复杂混合物。

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