NADPH oxidase of neutrophils elevates o,o′-dityrosine cross-links in proteins and urine during inflammation
NADPH oxidase of neutrophils elevates o,o′-dityrosine cross-links in proteins and urine during inflammation
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DOI：
10.1006/abbi.2001.2557
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发表时间：
2001-11-01
影响因子：
3.9
通讯作者：
Heinecke, JW
中科院分区：
文献类型：
--
作者：
Bhattacharjee, S;Pennathur, S;Heinecke, JW
吞噬性白细胞产生的活性中间体在消灭微生物方面至关重要，但它们也可能损伤炎症部位的正常组织。为了研究这类氧化剂在组织损伤中的潜在作用，我们使用气相色谱/质谱法来定量测定小鼠腹腔中性粒细胞和尿液中邻，邻’ - 二酪氨酸的水平。在野生型动物中，中性粒细胞在体内被激活时，其蛋白质结合的二酪氨酸含量显著增加。在吞噬细胞NADPH氧化酶缺陷的小鼠中，这种增加并未出现。尿液中邻，邻’ - 二酪氨酸的水平与中性粒细胞蛋白质中的水平相符。当将邻，邻’ - [C - 14]二酪氨酸静脉注射到小鼠体内时，放射性标记物没有被代谢或掺入组织蛋白质中，而是几乎以定量的产率从尿液中回收。脓毒症患者尿液中邻，邻’ - 二酪氨酸的排出量显著增加，这表明全身性炎症在人类中也可能是氧化应激的一个重要来源。这些观察结果表明，活化的中性粒细胞在组织蛋白质中产生邻，邻’ - 二酪氨酸交联，这些交联随后可能被降解为游离氨基酸并排泄到尿液中。我们的研究结果表明，小鼠吞噬细胞利用NADPH氧化酶产生的氧化剂在体内产生邻，邻’ - 二酪氨酸交联，并提出了通过该途径产生的活性中间体促进人类炎症性组织损伤的可能性。（C）2001年学术出版社
Reactive intermediates generated by phagocytic white blood cells are of central importance in destroying microorganisms, but they may also damage normal tissue at sites of inflammation. To investigate the potential role of such oxidants in tissue injury, we used gas chromatography/mass spectrometry to quantify levels of o,o ' -dityrosine in mouse peritoneal neutrophils and urine. In wild-type animals, neutrophils markedly increased their content of protein-bound dityrosine when they were activated in vivo. This increase failed to occur in mice that were deficient in the phagocyte NADPH oxidase. Levels of o,o ' -dityrosine in urine mirrored those in neutrophil proteins. When o,o '-[C-14]dityrosine was injected intravenously into mice, the radiolabel was not metabolized or incorporated into tissue proteins: instead, it was recovered in urine with near-quantitative yield. Patients with sepsis markedly increased their output of o,o ' -dityrosine into urine, suggesting that systemic inflammation also may be a potent source of oxidative stress in humans. These observations demonstrate that activated neutrophils produce o,o ' -dityrosine cross-links in tissue proteins, which may subsequently be degraded into free amino acids and excreted into urine. Our results indicate that mouse phagocytes use oxidants produced by the NADPH oxidase to create o,o ' -dityrosine cross-links in vivo and raise the possibility that reactive intermediates produced by this pathway promote inflammatory tissue damage in humans. (C) 2001 Academic Press.