PEROXYNITRITE FORMATION FROM MACROPHAGE-DERIVED NITRIC-OXIDE
PEROXYNITRITE FORMATION FROM MACROPHAGE-DERIVED NITRIC-OXIDE
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DOI：
10.1016/0003-9861(92)90433-w
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发表时间：
1992-11-01
影响因子：
3.9
通讯作者：
BECKMAN, JS
中科院分区：
文献类型：
--
作者：
ISCHIROPOULOS, H;ZHU, L;BECKMAN, JS
用Cu、Zn超氧化物歧化酶催化对羟基苯乙酸的硝化反应，测定了佛波醇12-肉豆蔻酸酯13-乙酸酯激活的大鼠肺泡巨噬细胞生成过氧亚硝酸根的能力。一氧化氮合成抑制剂N-甲基-L-精氨酸抑制了巨噬细胞对Cu，Zn超氧化物歧化酶催化的4-羟基苯乙酸硝化反应，而去Cu的Zn超氧化物歧化酶在体外和活化的巨噬细胞存在下均不催化3-硝基-4-羟基苯乙酸的生成。活化巨噬细胞的酚硝化速率为9 ± 2 pmol ·106 cells −1· min−1（平均值±标准差）。只有8%的合成过氧亚硝酸盐被超氧化物歧化酶捕获，这表明过氧亚硝酸盐的形成速率可能高达0.11 nmol ·106 cells −1· min−1。这一上限估计值与N-甲基-L-精氨酸使细胞色素c检测到的超氧化物的量增加0.12 nmol ·106 cells −1· min−1的结果一致。亚硝酸盐和硝酸盐的积累速率为0.10 ± 0.001 nmol ·106 cells −1· min−1，表明活化的巨噬细胞产生的大部分一氧化氮可能已经转化为过氧亚硝酸盐。在活化的巨噬细胞中，一氧化氮和超氧化物反应形成相对长寿命的强氧化剂可能有助于炎症细胞介导的组织损伤。
Peroxynitrite formation by rat alveolar macrophages activated with phorbol 12-myristate 13-acetate was assayed by the Cu,Zn superoxide dismutase-catalyzed nitration of 4-hydroxyphenylacetate. The inhibitor of nitric oxide synthesisN-methyl-l-arginine prevented the Cu,Zn Superoxide dismutase-catalyzed nitration of 4-hydroxyphenylacetate by stimulated macrophages, while Cu-depleted Zn superoxide dismutase did not catalyze the formation of 3-nitro-4-hydroxyphenylacetate eitherin vitroor in the presence of activated macrophages. The rate of phenolic nitration by activated macrophages was 9 ± 2 pmol · 106cells−1· min−1(mean ± STD). Only 8% of synthetic peroxynitrite was trapped by superoxide dismutase, which suggested that the rate of peroxynitrite formation may have been as high as 0.11 nmol · 106cells−1· min−1. This upper estimate was consistent withN-methyl-l-arginine increasing the amount of superoxide detected with cytochrome c by 0.12 nmol · 106cells−1· min−1. The rate of nitrite and nitrate accumulation was 0.10 ± 0.001 nmol · 106cells−1· min−1, suggesting that the majority of nitric oxide produced by activated macrophages may have been converted to peroxynitrite. The formation of a relatively long lived, strong oxidant from the reaction of nitric oxide and superoxide in activated macrophages may contribute to inflammatory cell-mediated tissue injury.