KINETICS AND SITE SPECIFICITY OF HYDROPEROXIDE-INDUCED OXIDATIVE DAMAGE IN RED-BLOOD-CELLS
KINETICS AND SITE SPECIFICITY OF HYDROPEROXIDE-INDUCED OXIDATIVE DAMAGE IN RED-BLOOD-CELLS
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DOI：
10.1016/0891-5849(92)90102-m
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发表时间：
1992-01-01
影响因子：
7.4
通讯作者：
KUYPERS, FA
中科院分区：
文献类型：
--
作者：
VANDENBERG, JJM;DENKAMP, JAF;KUYPERS, FA
为了提供一个详细的描述的时间过程和过氧化氢诱导的氧化应激在红细胞（RBC）的位点特异性，我们已经表征了膜溶性（氢过氧化枯烯[cumOOH]）和水溶性（过氧化氢[H2O2]）氧化剂的作用。荧光多不饱和脂肪酸（PUFA）parinaric酸（PnA）被用来探测过氧化过程中的膜，和血红蛋白（Hb）的氧化作为一个指标的胞质氧化应激进行了测定。观察到的降解模式的PnA和血红蛋白是明显不同的每种氧化剂。在可比的氧化剂浓度，RBC膜上的累积氧化应激总是高得多的cumOOH，而更多的Hb氧化与H2O2测量。Hb氧化的动力学以及所形成的产物的性质对于每种氧化剂是不同的。cumOOH逐渐氧化Hb生成的主要产物是方法，而H_2O_2导致快速生成铁酰Hb。CumOOH引起更多的内源性多不饱和脂肪酸和维生素E的氧化，而维生素E的降解模式非常相似的PnA。在高氧化剂浓度下，长时间孵育后观察到广泛的细胞溶解。丁基羟基甲苯（BHT）完全防止内源性PUFA的氧化，但不能完全防止溶血，表明脂质过氧化以外的因素也是导致红细胞溶解的重要因素。cumOOH的作用的特征在于与Hb逐渐反应，产生产生主要针对膜的氧化应激的自由基，其随时间增加至最大值，然后逐渐降低。相比之下，H2O2穿过RBC膜并与Hb快速反应，产生具有Hb而不是膜作为主要目标的非常活性的自由基物质。H2O2诱导的氧化应激在加入该氧化剂后立即达到最大值，并在短时间内迅速降至零。这些研究结果提供了进一步了解氢过氧化物的作用方式和红细胞氧化损伤的区室化机制。
To provide a detailed description of the time course and the site specificity of hydroperoxide-induced oxidative stress in red blood cells (RBCs), we have characterized the action of a membrane-soluble (cumene hydroperoxide [cumOOH]) and a water-soluble (hydrogen peroxide [H2O2]) oxidant. The fluorescent polyunsaturated fatty acid (PUFA) parinaric acid (PnA) was used to probe peroxidation processes in the membrane, and oxidation of hemoglobin (Hb) was measured spectrophotometrically as an indicator of cytosolic oxidative stress. The observed degradation patterns of PnA and Hb were clearly distinct for each oxidant. At comparable oxidant concentrations, the cumulative oxidative stress on the RBC membrane was always much higher with cumOOH, whereas much more Hb oxidation was measured with H2O2. The kinetics of Hb oxidation as well as the nature of the products formed were different for each oxidant. The main Hb oxidation product generated gradually by cumOOH was method, whereas H2O2 caused the rapid formation of ferrylHb. CumOOH caused more oxidation of endogenous PUFAs and of vitamin E, while the degradation pattern of vitamin E closely resembled that of PnA. At high oxidant concentrations, extensive cell lysis was observed after prolonged incubation. Butylated hydroxytoluene (BHT) completely prevented oxidation of endogenous PUFAs but did not completely prevent hemolysis, indicating that factors other than lipid peroxidation are also important in causing lysis of RBCs. The action of cumOOH is characterized by a gradual reaction with Hb, generating radicals that produce an oxidative stress primarily directed at the membrane, which increases in time to a maximum and then gradually decreases. In contrast, H2O2 crosses the RBC membrane and reacts rapidly with Hb, generating a very reactive radical species that has Hb, not the membrane, as a prime target. H2O2-induced oxidative stress is at a maximum immediately after addition of this oxidant and decreases rapidly to zero in a short time. These findings provide further insight into the mode of action of hydroperoxides and the mechanism of compartmentalization of RBC oxidative damage.