Reactions of nitric oxide and peroxynitrite with organic molecules and ferrihorseradish peroxidase: Interference with the determination of hydrogen peroxide
Reactions of nitric oxide and peroxynitrite with organic molecules and ferrihorseradish peroxidase: Interference with the determination of hydrogen peroxide
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DOI：
10.1016/0891-5849(95)02098-5
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发表时间：
1996-01-01
影响因子：
7.4
通讯作者：
AlMehdi, A
中科院分区：
文献类型：
--
作者：
Ischiropoulos, H;Nelson, J;AlMehdi, A
内皮细胞、炎性细胞和其他类型的细胞，在适当的刺激下，除了形成活性氧物种外，还会释放一氧化氮((NO)-N-)等自由基。以及强氧化剂，如过氧亚硝酸盐(ONOO-)，它是一氧化氮和超氧化物反应的产物。几种检测过氧化氢的方法都是基于铁辣根过氧化物酶催化的有机分子的氧化。我们研究了一氧化氮和过氧亚硝酸盐与有机分子以及与辣根过氧化物酶的反应，并考察了它们对H_2O_2检测的潜在干扰。低浓度的过氧亚硝酸根(0-10 mU M)对1，2-苯二胺、3-3‘-二甲氧基联苯胺(o-dianisidine)和对羟基苯乙酸(PHPA)具有浓度依赖性的氧化作用。除PHPA外，过氧亚硝酸根对上述化合物的氧化均不受辣根过氧化物酶的影响。由于辣根过氧化物酶催化生成不同的产物3-硝基-HPA，导致过亚硝酸根氧化产物HPA-二聚体的产率降低。一氧化氮与氧气反应生成的氮氧化物氧化芳香胺邻苯二胺和邻二苯二胺。与过氧化氢加辣根过氧化物酶相比，过量10倍的一氧化氮使PHPA和二氢罗丹明123(DHR123)的产率分别下降58%和72%。一氧化氮对PHPA氧化的抑制作用部分是由于一氧化氮与化合物I和化合物II的反应，部分是由于与苯氧基自由基的反应。这些数据表明，一氧化氮和过氧亚硝酸盐的同时产生可能会干扰过氧化氢的检测。对检测到的过氧化氢的高估或低估取决于用于检测的有机分子以及一氧化氮、超氧化物和过氧亚硝酸盐生成的相对速率。
Endothelial, inflammatory, and other cell types, in addition to forming reactive oxygen species, under proper stimulation release free radicals such as nitric oxide ((NO)-N-.) and strong oxidants such as peroxynitrite (ONOO-), which is the product of the reaction of nitric oxide with superoxide. Several methods for the detection of H2O2 are based on the ferrihorseradish peroxidase catalyzed oxidation of organic molecules. We investigated the reactions of nitric oxide and peroxynitrite with organic molecules as well as with ferrihorseradish peroxidase and examined the potential interference with the detection of H2O2. Peroxynitrite at low concentrations (0-10 mu M) induced a concentration-dependent oxidation of 1,2 phenylenediamine, 3-3' dimethoxybenzidine (o-dianisidine) and para-hydroxyphenylacetic acid (pHPA). With the exception of pHPA, the oxidation of the above compounds by peroxynitrite was not affected by the presence of ferrihorseradish peroxidase. The yield of HPA-dimmer, the oxidation product of pHPA by peroxynitrite, was decreased because ferrihorseradish peroxidase catalyzed the formation of a different product, 3-nitro-HPA. Nitrogen oxides, formed by the reaction of nitric oxide with oxygen, oxidized the aromatic amines o-phenylenediamine and o-dianisidine. A 10-fold excess of nitric oxide over H2O2 decreased the yield of pHPA and dihydrorhodamine 123 (DHR123) by 58 and 72%, respectively, as compared to H2O2 plus ferrihorseradish peroxidase. The inhibition of pHPA oxidation by nitric oxide was due in part to the reaction of nitric oxide with compound I and compound II and in part due to the reaction with the phenoxyl radical. These data suggest that the simultaneous generation of nitric oxide and peroxynitrite can interfere with the detection of H2O2. The overestimation or underestimation of the H2O2 detected is dependent upon the organic molecule utilized for detection and by the relative rate of nitric oxide, superoxide, and peroxynitrite generation.