Nurkhametova Dilyara, Kudryavtsev Igor, Guselnikova Valeriia, Serebryakova Maria, Giniatullina Raisa R, Wojciechowski Sara, Tore Fatma, Rizvanov Albert, Koistinaho Jari, Malm Tarja, Giniatullin Rashid
Laboratory of Neurobiology, Kazan Federal University, Kazan, Russia.
A.I.Virtanen Institute for Molecular Sciences, University of Eastern Finland, Kuopio, Finland.
Front Cell Neurosci. 2019 Feb 13;13:45. doi: 10.3389/fncel.2019.00045. eCollection 2019.
Extracellular ATP activates inflammasome and triggers the release of multiple cytokines in various immune cells, a process primarily mediated by P2X7 receptors. However, the expression and functional properties of P2X7 receptors in native mast cells in tissues such as meninges where migraine pain originates from have not been explored. Here we report a novel model of murine cultured meningeal mast cells and using these, as well as easily accessible peritoneal mast cells, studied the mechanisms of ATP-mediated mast cell activation. We show that ATP induced a time and dose-dependent activation of peritoneal mast cells as analyzed by the uptake of organic dye YO-PRO1 as well as 4,6-diamidino-2-phenylindole (DAPI). Both YO-PRO1 and DAPI uptake in mast cells was mediated by the P2X7 subtype of ATP receptors as demonstrated by the inhibitory effect of P2X7 antagonist A839977. Consistent with this, significant YO-PRO1 uptake was promoted by the P2X7 agonist 2',3'-O-(benzoyl-4-benzoyl)-ATP (BzATP). Extracellular ATP-induced degranulation of native and cultured meningeal mast cells was shown with Toluidine Blue staining. Taken together, these data demonstrate the important contribution of P2X7 receptors to ATP-driven activation of mast cells, suggesting these purinergic mechanisms as potential triggers of neuroinflammation and pain sensitization in migraine.
细胞外ATP可激活炎性小体并触发多种免疫细胞中多种细胞因子的释放，这一过程主要由P2X7受体介导。然而，偏头痛疼痛起源的组织（如脑膜）中天然肥大细胞中P2X7受体的表达和功能特性尚未得到研究。在此，我们报告了一种小鼠培养脑膜肥大细胞的新模型，并利用这些细胞以及易于获取的腹膜肥大细胞，研究了ATP介导的肥大细胞活化机制。我们发现，通过有机染料YO-PRO1以及4,6-二脒基-2-苯基吲哚（DAPI）的摄取分析，ATP诱导了腹膜肥大细胞的时间和剂量依赖性活化。肥大细胞中YO-PRO1和DAPI的摄取均由ATP受体的P2X7亚型介导，这一点通过P2X7拮抗剂A839977的抑制作用得以证明。与此一致的是，P2X7激动剂2',3'-O-（苯甲酰基-4-苯甲酰基）-ATP（BzATP）促进了显著的YO-PRO1摄取。通过甲苯胺蓝染色显示，细胞外ATP诱导了天然和培养的脑膜肥大细胞脱颗粒。综上所述，这些数据证明了P2X7受体对ATP驱动的肥大细胞活化的重要贡献，表明这些嘌呤能机制可能是偏头痛中神经炎症和疼痛敏化的触发因素。
