Roberge C J, Gaudry M, de Médicis R, Lussier A, Poubelle P E, Naccache P H
Centre de Recherche en Inflammation, Immunologie et Rhumatologie, Université Laval, Québec, Canada.
J Clin Invest. 1993 Oct;92(4):1722-9. doi: 10.1172/JCI116759.
We recently demonstrated that pathologically relevant inflammatory microcrystals, namely triclinic monosodium urate (MSU) and calcium pyrophosphate dihydrate (CPPD) crystals, potently stimulate a characteristic protein tyrosine phosphorylation pattern in human neutrophils that differed from that observed in response to other soluble or particulate agonists. In this study, the effects of colchicine on protein tyrosine phosphorylation induced by MSU and CPPD crystals in human blood neutrophils were investigated. Immunoblot analysis with antiphosphotyrosine antibodies demonstrated that colchicine dose-dependently inhibited the tyrosine phosphorylation of all the proteins phosphorylated in response to MSU and CPPD crystals. Other microtubule-disruptive agents such as vinblastine, nocodazole, and colcemid also inhibited crystal-induced protein tyrosine phosphorylation while lumicolchicine and trimethylcolchicinic acid were without effect. Indomethacin and phenylbutazone were similarly without effect on microcrystal-induced tyrosine phosphorylation. Colchicine, as well as the other active alkaloids, failed to inhibit the protein tyrosine phosphorylation elicited by FMLP, C5a, leukotriene B4, and unopsonized zymosan. Overall, these results demonstrate that colchicine specifically and significantly inhibits the protein tyrosine phosphorylation induced by MSU and CPPD crystals and suggest that its effects are associated, at least in part, with its interaction with microtubules. Furthermore, the use of microtubule-disrupting drugs demonstrate that the mechanisms implicated in the induction of protein tyrosine phosphorylation by microcrystals differed from those involved in response to other soluble or particulate agonists.
我们最近证明，具有病理相关性的炎性微晶，即三斜晶型尿酸钠（MSU）和二水焦磷酸钙（CPPD）晶体，能有效刺激人类中性粒细胞中一种特征性的蛋白酪氨酸磷酸化模式，该模式不同于对其他可溶性或颗粒性激动剂的反应所观察到的模式。在本研究中，研究了秋水仙碱对人血中性粒细胞中MSU和CPPD晶体诱导的蛋白酪氨酸磷酸化的影响。用抗磷酸酪氨酸抗体进行的免疫印迹分析表明，秋水仙碱剂量依赖性地抑制了所有因MSU和CPPD晶体而磷酸化的蛋白的酪氨酸磷酸化。其他破坏微管的药物，如长春碱、诺考达唑和秋水仙酰胺，也抑制晶体诱导的蛋白酪氨酸磷酸化，而光秋水仙碱和三甲基秋水仙碱则无此作用。吲哚美辛和保泰松同样对微晶诱导的酪氨酸磷酸化无影响。秋水仙碱以及其他活性生物碱未能抑制由FMLP、C5a、白三烯B4和未调理的酵母聚糖引起的蛋白酪氨酸磷酸化。总体而言，这些结果表明秋水仙碱能特异性且显著地抑制MSU和CPPD晶体诱导的蛋白酪氨酸磷酸化，并提示其作用至少部分与其与微管的相互作用有关。此外，破坏微管药物的使用表明，微晶诱导蛋白酪氨酸磷酸化的机制不同于对其他可溶性或颗粒性激动剂反应所涉及的机制。
