Evidence for in vivo peroxynitrite production in human acute lung injury.
Evidence for in vivo peroxynitrite production in human acute lung injury.
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DOI：
10.1164/ajrccm/151.4.1250
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发表时间：
1995-04
影响因子：
24.7
通讯作者：
N. Kooy;J. Royall;Y. Ye;D. R. Kelly;J. Beckman
中科院分区：
文献类型：
--
作者：
N. Kooy;J. Royall;Y. Ye;D. R. Kelly;J. Beckman
急性肺部炎症引起的氧化介导毒性已在急性肺损伤中得到证实。一种强效的生物氧化剂，过氧亚硝酸盐，是由一氧化氮与超氧化物的近扩散限制反应形成的。除了具有类似羟基自由基的氧化反应活性外，过氧亚硝酸盐还能够硝化酚环，包括蛋白质相关的酪氨酸残基。一氧化氮不直接使酪氨酸残基硝酸盐化，因此，组织硝基酪氨酸残基的发现推断出过氧亚硝酸盐或相关氮中心氧化剂的作用。解剖标本经福尔马林固定、石蜡包埋后获得肺组织，通过二氨基苯-过氧化物酶染色观察到针对硝基酪氨酸的特异性多克隆和单克隆抗体。急性肺损伤导致全肺强烈染色，包括肺间质、肺泡上皮、肺泡蛋白渗出物和炎症细胞。此外，脓毒症致急性肺损伤患者的血管内皮和内皮下组织出现染色。与硝基酪氨酸或硝化牛血清白蛋白共孵育可阻断抗体结合，而与氨基酪氨酸、磷酸酪氨酸或牛血清白蛋白共孵育则不能阻断抗体结合。氢亚硫酸钠将组织硝基酪氨酸还原为氨基酪氨酸也阻断了抗体的结合。在没有明显肺部疾病的对照标本中，只有轻微的肺泡隔染色。这些结果表明，氮源氧化剂在人类急性肺损伤中形成，并提示过氧亚硝酸盐可能是炎症性肺病的重要氧化剂。
Oxidant-mediated toxicity resulting from acute pulmonary inflammation has been demonstrated in acute lung injury. A potent biological oxidant, peroxynitrite, is formed by the near diffusion-limited reaction of nitric oxide with superoxide. In addition to having hydroxyl radical-like oxidative reactivity, peroxynitrite is capable of nitrating phenolic rings, including protein-associated tyrosine residues. Nitric oxide does not directly nitrate tyrosine residues, therefore, demonstration of tissue nitrotyrosine residues infers the action of peroxynitrite or related nitrogen-centered oxidants. Lung tissue was obtained from formalin-fixed, paraffin-embedded autopsy specimens, and specific polyclonal and monoclonal antibodies to nitrotyrosine were visualized by diaminobenzidene-peroxidase staining. Acute lung injury resulted in intense staining throughout the lung, including lung interstitium, alveolar epithelium, proteinaceous alveolar exudate, and inflammatory cells. In addition, staining of the vascular endothelium and subendothelial tissues was present in those patients with sepsis-induced acute lung injury. Antibody binding was blocked by coincubation with nitrotyrosine or nitrated bovine serum albumin but not by aminotyrosine, phosphotyrosine, or bovine serum albumin. Reduction of tissue nitrotyrosine to aminotyrosine by sodium hydrosulfite also blocked antibody binding. In control specimens with no overt pulmonary disease, there was only slight staining of the alveolar septum. These results demonstrate that nitrogen-derived oxidants are formed in human acute lung injury and suggest that peroxynitrite may be an important oxidant in inflammatory lung disease.