Bednar M M, Balazy M, Murphy M, Booth C, Fuller S P, Barton A, Bingham J, Golding L, Gross C E
Division of Neurosurgery, University of Vermont, Burlington 05405-0068, USA.
J Leukoc Biol. 1996 Nov;60(5):619-24. doi: 10.1002/jlb.60.5.619.
The neutrophil respiratory burst was examined by the technique of luminol-dependent chemiluminescence (LDCL) triggered by submaximal concentrations of N-formyl-methionyl-leucyl-phenylalanine (fMLP) in diluted whole blood. We sought to identify the chemical species responsible for LDCL in whole blood, to examine the role of leukotriene B4 (LTB4) and other arachidonic acid metabolites as mediators of the fMLP signaling pathway, and to investigate the effect of peroxynitrite on this response. Both sodium azide and taurine significantly inhibited LDCL (93% inhibition with 100 microM azide, 52% inhibition with 10 mM taurine). More modest inhibition was seen with superoxide dismutase (SOD), catalase, the nitric oxide synthase inhibitor monomethyl-L-arginine (L-NMMA), and with inhibitors of the cyclooxygenase (indomethacin), lipoxygenase (AA-861; no effect), and cytochrome P-450 (SKF 525-A) pathways of arachidonic acid metabolism. The nitric oxide donor SIN-1 (1-100 microM) and peroxynitrite (10-300 microM) also augmented fMLP-induced LDCL. The augmentation seen with peroxynitrite and SIN-1 was attenuated by SOD. Despite the increase in LDCL, peroxynitrite caused a dose-related inhibition of fMLP-stimulated LTB4 release. In summary, our results indicate that (1) LDCL elicited by fMLP in diluted whole blood appears primarily mediated by hypochlorous acid derived from myeloperoxidase; (2) pretreatment with the nitric oxide donor SIN-1 or with peroxynitrite augments LDCL; and (3) LTB4 release does not contribute to fMLP-stimulated LDCL or in the modulation of LDCL by SIN-1 or peroxynitrite.
通过在稀释全血中用亚最大浓度的N-甲酰甲硫氨酰-亮氨酰-苯丙氨酸（fMLP）触发的鲁米诺依赖性化学发光（LDCL）技术检测中性粒细胞呼吸爆发。我们试图确定全血中负责LDCL的化学物质，研究白三烯B4（LTB4）和其他花生四烯酸代谢产物作为fMLP信号通路介质的作用，并研究过氧亚硝酸盐对该反应的影响。叠氮化钠和牛磺酸均显著抑制LDCL（100μM叠氮化物抑制93%，10mM牛磺酸抑制52%）。超氧化物歧化酶（SOD）、过氧化氢酶、一氧化氮合酶抑制剂单甲基-L-精氨酸（L-NMMA）以及花生四烯酸代谢的环氧化酶（吲哚美辛）、脂氧化酶（AA-861；无作用）和细胞色素P-450（SKF 525-A）途径的抑制剂产生的抑制作用较弱。一氧化氮供体SIN-1（1-100μM）和过氧亚硝酸盐（10-300μM）也增强了fMLP诱导的LDCL。过氧亚硝酸盐和SIN-1引起的增强作用被SOD减弱。尽管LDCL增加，但过氧亚硝酸盐导致fMLP刺激的LTB4释放呈剂量依赖性抑制。总之，我们的结果表明：（1）fMLP在稀释全血中引发的LDCL似乎主要由髓过氧化物酶衍生的次氯酸介导；（2）用一氧化氮供体SIN-1或过氧亚硝酸盐预处理可增强LDCL；（3）LTB4释放对fMLP刺激的LDCL或SIN-1或过氧亚硝酸盐对LDCL的调节无贡献。
