Gaudry M, Roberge C J, de Médicis R, Lussier A, Poubelle P E, Naccache P H
Centre de Recherche en Inflammation et Immunologie-Rhumatologie, Université Laval, Québec, Canada.
J Clin Invest. 1993 Apr;91(4):1649-55. doi: 10.1172/JCI116373.
The activation of human neutrophils by monosodium urate and calcium pyrophosphate dihydrate crystals is believed to play a critical role in the pathogenesis of arthritides such as acute gout and pseudogout, respectively. In this study, we investigated the potential involvement of tyrosine phosphorylation in microcrystal-mediated activation of human neutrophils. Immunoblot analysis with antiphosphotyrosine antibodies demonstrated that triclinic monosodium urate and calcium pyrophosphate dihydrate crystals stimulated a time- and concentration-dependent tyrosine phosphorylation of at least five proteins (pp130, 118, 80, 70, and 60). While phosphoprotein (pp) 118 and pp70 were the major phosphorylated substrates, pp70 was the dominant one in reactivity with antiphosphotyrosine antibodies. When the temporal patterns, as well as the levels of tyrosine phosphorylation for both types of crystals were compared, monosodium urate crystals were found to be more potent activators than calcium pyrophosphate dihydrate crystals. The tyrosine phosphorylation patterns induced by microcrystals differed from those stimulated by other soluble (FMLP, C5a, or leukotriene B4) or particulate (unopsonized latex beads or zymosan) agonists which stimulated preferentially the tyrosine phosphorylation of pp118. The ratio of the intensities of pp118 and pp70 were specific of the stimulation with microcrystals when compared to those observed with the other soluble or particulate agonists. Colchicine, a drug used specifically in the treatment of gout and pseudogout, inhibited microcrystal-induced tyrosine phosphorylation, while beta- and gamma-lumicolchicine were without effect. On the other hand, colchicine failed to inhibit FMLP-induced tyrosine phosphorylation. Furthermore, while colchicine inhibited the activation of the NADPH oxidase by microcrystals, it, on the other hand, enhanced the production of superoxide anions by FMLP. Taken together, these results (a) demonstrate that tyrosine phosphorylation is involved in the mechanism of activation of human neutrophils induced by microcrystals; and (b) suggest, on the basis of the characteristics of the observed patterns of tyrosine phosphorylation, that this response may be specific to the microcrystals and relevant to their phlogistic properties.
尿酸单钠和二水焦磷酸钙晶体激活人类中性粒细胞，分别被认为在急性痛风和假性痛风等关节炎的发病机制中起关键作用。在本研究中，我们调查了酪氨酸磷酸化在微晶介导的人类中性粒细胞激活中的潜在作用。用抗磷酸酪氨酸抗体进行免疫印迹分析表明，三斜晶型尿酸单钠和二水焦磷酸钙晶体刺激了至少五种蛋白质（pp130、118、80、70和60）的时间和浓度依赖性酪氨酸磷酸化。虽然磷蛋白（pp）118和pp70是主要的磷酸化底物，但pp70是与抗磷酸酪氨酸抗体反应性最强的底物。当比较两种晶体的酪氨酸磷酸化的时间模式和水平时，发现尿酸单钠晶体比二水焦磷酸钙晶体更有效地激活细胞。微晶诱导的酪氨酸磷酸化模式与其他可溶性（FMLP、C5a或白三烯B4）或颗粒性（未调理的乳胶珠或酵母聚糖）激动剂刺激的模式不同，后者优先刺激pp118的酪氨酸磷酸化。与其他可溶性或颗粒性激动剂相比，pp118和pp70强度的比值是微晶刺激所特有的特征。秋水仙碱是一种专门用于治疗痛风和假性痛风的药物，它能抑制微晶诱导的酪氨酸磷酸化，而β-和γ-光秋水仙碱则没有作用。另一方面，秋水仙碱不能抑制FMLP诱导的酪氨酸磷酸化。此外，虽然秋水仙碱抑制了微晶对NADPH氧化酶的激活，但另一方面，它增强了FMLP诱导的超氧阴离子的产生。综上所述，这些结果（a）表明酪氨酸磷酸化参与了微晶诱导的人类中性粒细胞激活机制；（b）基于观察到的酪氨酸磷酸化模式的特征表明，这种反应可能是微晶所特有的，并且与其致炎特性相关。
