Colchicine inhibits cationic dye uptake induced by ATP in P2X2 and P2X7 receptor-expressing cells: implications for its therapeutic action
Colchicine inhibits cationic dye uptake induced by ATP in P2X2 and P2X7 receptor-expressing cells: implications for its therapeutic action
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DOI：
10.1111/j.1476-5381.2011.01254.x
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发表时间：
2011-07-01
影响因子：
7.3
通讯作者：
Guimaraes, M. Z. P.
中科院分区：
文献类型：
--
作者：
Marques-da-Silva, C.;Chaves, M. M.;Guimaraes, M. Z. P.
背景与目的嘌呤能P2X受体的两个最长的c端出现在P2X2和P2X7受体中，被认为与多种细胞质蛋白相互作用，其中包括细胞骨架的成员，包括微管。在这项工作中，我们询问破坏微管细胞骨架是否会影响这些受体的功能。实验方法应用三磷酸腺苷（ATP）对异源表达的P2X2和P2X7受体的功能进行电生理和染料摄取评价。对新鲜或培养的小鼠腹腔巨噬细胞进行体外或体内秋水仙碱处理，定量测定促炎剂的通透性和分泌。关键结果秋水仙碱破坏微管网络不影响P2X2和P2X7受体表达细胞中ATP产生的电流，但抑制了爪蟾卵母细胞和表达这些通道的HEK293细胞对Yo-Pro-1染料的摄取。在秋水仙碱存在的情况下，小鼠腹腔巨噬细胞表现出atp诱导的溴化乙啶通透性降低，活性氧（ROS）形成、一氧化氮（NO）和白细胞介素(IL)-1 β释放减少。秋水仙碱处理不影响巨噬细胞atp诱发电流。最后，小鼠体内实验显示，秋水仙碱处理后，小鼠体内ROS、IL-1 β、干扰素γ和NO的产生减少。结论和意义：鼠戊子碱具有已知的抗炎作用，可用于治疗几种涉及先天免疫的疾病，包括痛风和家族性地中海热。本文提出了一种新的作用机制——P2X受体激活诱导的孔形成抑制，这可以解释秋水仙碱的一些抗炎作用。这篇文章由Pelegrin评论，第908-911页。要查看此评论，请访问http://dx.doi.org/10.1111/j.1476-5381.2011.01325.x。
BACKGROUND AND PURPOSEThe two longest C-termini of the purinergic P2X receptors occur in the P2X2 and P2X7 receptors and are thought to interact with multiple cytoplasmic proteins, among which are members of the cytoskeleton, including microtubules. In this work we asked whether disrupting the microtubule cytoskeleton might affect the functions of these receptors.EXPERIMENTAL APPROACHFunctions of heterologously expressed P2X2 and P2X7 receptors were evaluated with electrophysiology and dye uptake following ATP application. Permeabilization and secretion of pro-inflammatory agents were quantified from fresh or cultured peritoneal mouse macrophages, treated in vitro or in vivo with colchicine.KEY RESULTSDisrupting the microtubule network with colchicine did not affect currents generated by ATP in P2X2 and P2X7 receptor-expressing cells but inhibited uptake of the dye Yo-Pro-1 in Xenopus oocytes and HEK293 cells expressing these channels. Peritoneal mouse macrophages showed less ATP-induced permeabilization to ethidium bromide in the presence of colchicine, and less reactive oxygen species (ROS) formation, nitric oxide (NO) and interleukin (IL)-1 beta release. Colchicine treatment did not affect ATP-evoked currents in macrophages. Finally, in vivo assays with mice inoculated with lipopolysaccharide and ATP showed diminished ROS, IL-1 beta, interferon-gamma and NO production after colchicine treatment.CONCLUSIONS AND IMPLICATIONSColchicine has known anti-inflammatory actions and is used to treat several conditions involving innate immunity, including gout and familial Mediterranean fever. Here we propose a new mechanism of action - inhibition of pore formation induced by activation of P2X receptors - which could explain some of the anti-inflammatory effects of colchicine.LINKED ARTICLEThis article is commented on by Pelegrin, pp. 908-911 of this issue. To view this commentary visit http://dx.doi.org/10.1111/j.1476-5381.2011.01325.x.