Peroxynitrite formation from activated human leukocytes
Peroxynitrite formation from activated human leukocytes
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DOI：
10.1006/bbrc.1996.1041
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发表时间：
1996-07-16
影响因子：
3.1
通讯作者：
Nakazawa, H
中科院分区：
文献类型：
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作者：
Fukuyama, N;Ichimori, K;Nakazawa, H
我们用4-羟基苯乙酸（HPA）硝化成4-羟基-3-硝基苯乙酸（NO（2）HPA）的方法，直接证明了多形核细胞（PMN）中过氧亚硝酸盐的形成。来自健康志愿者的人PMN用佛波醇-12-肉豆蔻酸酯-13-乙酸酯（PMA，10 ng/ml）在37 ℃下在含有HPA（1 mM）的2-[4-（2-羟乙基）-1-哌嗪基]乙磺酸缓冲的Hank平衡盐溶液（pH 7.4）中刺激。NO（2）HPA仅在有髓过氧化物酶抑制剂存在时才能在PMA刺激下被检测到。NO（2）HPA被N-单甲基-L-精氨酸（100 μ M）清除。髓过氧化物酶的抑制对于证明NO（2）HPA的产生似乎是必不可少的，因为髓过氧化物酶本身或其产物次氯酸盐与过氧亚硝酸盐反应并阻碍NO（2）HPA的形成。(C)出版社：Academic Press，Inc.
We showed direct evidence of peroxynitrite formation from polymorphonuclear cells (PMN) with the nitration of 4-hydroxpphenylacetic acid (HPA) to 4-hydroxy-3-nitrophenylacetic acid (NO(2)HPA). Human PMN from healthy volunteers was stimulated with phorbol-12-myristate-13-acetate (PMA, 10 ng/ml) at 37 degrees C in 2-[4-(2-hydroxyethyl)-1-piperazinyl]ethanesulfonic acid-buffered Hank' s balanced salt solution (pH 7.4) with HPA (1 mM). NO(2)HPA was detected under PMA stimulation only in the presence of myeloperoxidase inhibitor. NO(2)HPA was eliminated by N-monomethyl-L-arginine (100 mu M). The inhibition of myeloperoxidase appears to be essential to demonstrate the production of NO(2)HPA since myeloperoxidase itself or its product, hypochlorite, reacted with peroxynitrite and hampered the formation of NO(2)HPA. (C) 1996 Academic Press, Inc.