Sato E, Simpson K L, Grisham M B, Koyama S, Robbins R A
Research Services, Tucson and Overton Brooks VA Medical Centers, Department of Medicine, University of Arizona, Tucson, Arizona, USA.
Am J Pathol. 1999 Aug;155(2):591-8. doi: 10.1016/S0002-9440(10)65154-1.
Eosinophils and increased production of nitric oxide (NO) and superoxide, components of peroxynitrite, have been implicated in the pathogenesis of a number of allergic disorders including asthma. Peroxynitrite induced protein nitration may compromise enzyme and protein function. We hypothesized that peroxynitrite may modulate eosinophil migration by modulating chemotactic cytokines. To test this hypothesis, the eosinophil chemotactic responses of regulated on activation, normal T cell expressed and secreted (RANTES) and interleukin (IL)-5 incubated with and without peroxynitrite were evaluated. Peroxynitrite-attenuated RANTES and IL-5 induced eosinophil chemotactic activity (ECA) in a dose-dependent manner (P < 0.05) but did not attenuate leukotriene B4 or complement-activated serum ECA. The reducing agents deferoxamine and dithiothreitol reversed the ECA inhibition by peroxynitrite, and exogenous L-tyrosine abrogated the inhibition by peroxynitrite. PAPA-NONOate, a NO donor, or superoxide generated by lumazine or xanthine and xanthine oxidase, did not show an inhibitory effect on ECA. The peroxynitrite generator, 3-morpholinosydnonimine, caused a concentration-dependent inhibition of ECA. Peroxynitrite reduced RANTES and IL-5 binding to eosinophils and resulted in nitrotyrosine formation. These findings are consistent with nitration of tyrosine by peroxynitrite with subsequent inhibition of RANTES and IL-5 binding to eosinophils and suggest that peroxynitrite may play a role in regulation of eosinophil chemotaxis.
嗜酸性粒细胞以及过氧亚硝酸盐的成分一氧化氮（NO）和超氧化物产量增加，与包括哮喘在内的多种过敏性疾病的发病机制有关。过氧亚硝酸盐诱导的蛋白质硝化可能损害酶和蛋白质功能。我们假设过氧亚硝酸盐可能通过调节趋化细胞因子来调节嗜酸性粒细胞迁移。为了验证这一假设，评估了在有或无过氧亚硝酸盐的情况下，与调节激活正常T细胞表达和分泌因子（RANTES）和白细胞介素（IL）-5一起孵育时嗜酸性粒细胞的趋化反应。过氧亚硝酸盐以剂量依赖的方式减弱RANTES和IL-5诱导的嗜酸性粒细胞趋化活性（ECA）（P<0.05），但不减弱白三烯B4或补体激活血清ECA。还原剂去铁胺和二硫苏糖醇可逆转过氧亚硝酸盐对ECA的抑制作用，外源性L-酪氨酸可消除过氧亚硝酸盐的抑制作用。NO供体PAPA-NONOate或由鲁米诺或黄嘌呤及黄嘌呤氧化酶产生的超氧化物对ECA没有抑制作用。过氧亚硝酸盐生成剂3-吗啉代 sydnonimine引起ECA的浓度依赖性抑制。过氧亚硝酸盐减少RANTES和IL-5与嗜酸性粒细胞的结合，并导致硝基酪氨酸形成。这些发现与过氧亚硝酸盐使酪氨酸硝化，随后抑制RANTES和IL-5与嗜酸性粒细胞结合一致，并表明过氧亚硝酸盐可能在嗜酸性粒细胞趋化调节中发挥作用。
