Nitric oxide and nitrotyrosine in the lungs of patients with acute respiratory distress syndrome
Nitric oxide and nitrotyrosine in the lungs of patients with acute respiratory distress syndrome
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DOI：
10.1164/ajrccm.163.2.2004187
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发表时间：
2001-02-01
影响因子：
24.7
通讯作者：
Martin, TR
中科院分区：
文献类型：
--
作者：
Sittipunt, C;Steinberg, KP;Martin, TR
一氧化氮（NO）终产物（硝酸盐和亚硝酸盐）存在于炎症性肺病患者的支气管肺泡灌洗液（BAL）中。活性氧-氮中间体通过蛋白质中关键残基的氧化或硝化破坏大分子。本研究的目的是测量急性呼吸窘迫综合征（ARDS）发作前后支气管肺泡灌洗液中的NO终产物（硝酸盐+亚硝酸盐），并确定这些产物是否与诱导型一氧化氮合酶（iNOS）的表达相关。在BAL细胞中和BAL蛋白的硝化。我们在发病后第1、3、7、14和21天对有ARDS风险的患者（n = 19）或ARDS患者（n = 41）进行支气管肺泡灌洗（BAL），并测量每个BAL液样本中的总亚硝酸盐（将硝酸盐还原为亚硝酸盐后）和蛋白相关硝基酪氨酸浓度。通过免疫细胞化学分析BAL细胞的细胞离心液制备物的iNOS和硝基酪氨酸。硝酸盐+亚硝酸盐检测BAL液中的患者在危险的ARDS，并长达21天后的ARDS发作。在ARDS发作后长达14天的所有BAL液样本中均可检测到硝基酪氨酸（范围为38.8至278.5 pmol/mg蛋白质），但在正常志愿者的BAL中未检测到。急性呼吸窘迫综合征患者肺泡巨噬细胞iNOS和硝基酪氨酸均呈阳性表达，并在发病后14 d内仍呈阳性表达。BAL硝酸盐+亚硝酸盐不能预测ARDS的发生，但在死亡的患者中，在ARDS的第3天和第7天的浓度显著较高。因此，在ARDS发作之前和之后，NO终产物在肺中积累;在ARDS期间，iNOS在AM中以高水平表达;并且在ARDS中，细胞内和细胞外蛋白质的硝化发生在肺中。数据支持的概念，一氧化氮依赖性途径是重要的，在肺的患者之前和之后的ARDS发作。
Nitric oxide (NO) end-products (nitrate and nitrite) are present in bronchoalveolar lavage (BAL) fluid of patients with inflammatory lung diseases. Reactive oxygen-nitrogen intermediates damage macromolecules by oxidation or nitration of critical residues in proteins. The goal of this study was to measure NO end-products (nitrate + nitrite), in BAL fluid before and after the onset of acute respiratory distress syndrome (ARDS) and to determine if these products are associated with expression of inducible nitric oxide synthase enzyme (iNOS) in BAL cells and nitration of BAL proteins. We performed bronchoalveolar lavage (BAL) in patients at risk for ARDS (n = 19), or with ARDS (n = 41) on Days 1, 3, 7, 14, and 21 after onset, and measured total nitrite (after reducing nitrate to nitrite) and protein-associated nitrotyrosine concentration in each BAL fluid sample. Cytospin preparations of BAL cells were analyzed by immunocytochemistry for iNOS and nitrotyrosine. Nitrate + nitrite were detected in BAL fluid from patients at risk for ARDS, and for as long as 21 d after the onset of ARDS. Nitrotyrosine was detectable in all BAL fluid samples for as long as 14 d after the onset of ARDS (range, 38.8 to 278.5 pmol/mg of protein), but not in BAL of normal volunteers. Alveolar macrophages of patients with ARDS were positive for iNOS and nitrotyrosine, and remained positive for as long as 14 d after onset of ARDS. The BAL nitrate + nitrite did not predict the onset of ARDS, but the concentration was significantly higher on Days 3 and 7 of ARDS in patients who died. Thus, NO end products accumulate in the lungs before and after onset of ARDS; iNOS is expressed at high levels in AM during ARDS; and nitration of intracellular and extracellular proteins occurs in the lungs in ARDS. The data support the concept that NO-dependent pathways are important in the lungs of patients before and after the onset of ARDS.