Purinergic signaling in endothelial cells.

内皮细胞中的嘌呤能信号传导。

• 批准号: 6624238
• 负责人: ELZBIETA KACZMAREK
• 金额: $ 25.5万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-04-01 至 2006-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6624238
• 关键词: apoptosis; biological signal transduction; calcium flux; caveolins; cell proliferation; enzyme linked immunosorbent assay; focal adhesion kinase; genetically modified animals; inflammation; laboratory mouse; leukocyte activation /transformation; mitogen activated protein kinase; nuclear factor kappa beta; nucleosides; paxillin; phosphorylation; purinergic receptor; receptor expression; selectins; tissue /cell culture; vascular endothelium; western blottings

Extracellular tri- and diphosphate nucleosides are released into tissue fluids and plasma as a consequence of cellular responses to various pro- inflammatory stimuli, tissue damage and cell death. Extracellular nucleotides exert their effects on various cells through purinergic P2 type receptors and can cause cell activation and apoptosis. Our data show that extracellular ATP induces endothelial cell (EC) activation and phosphorylation of several proteins in EC, including focal adhesion kinase (FAK), paxillin, related adhesion focal tyrosine kinase (RAFTK), p130/cas, caveolin-1,Shc, and p38, SAPK/JNK and ERK MAP kinases. These proteins belong to various signal transduction pathways associated with cell cytoskeleton rearrangements, cell spreading, motility, cell proliferation, and apoptosis. We also found that extracellular nucleotides activate nuclear factor kappa B (NF-kappaB) and up-regulated expression of E-selectin, involved in the initial interaction of leukocytes with EC, leading to their transmigration into spots of inflammation. We intend to identify the receptor(s) responsible for up-regulation of the E- selectin and elucidate the mechanism of ATP-induced signal transduction leading to induction of E-selectin gene. Obtained results will help to elucidate the mechanisms of disease states that involve purinergic receptor signaling (e.g, inflammation, angiogenesis, atherogenesis, graft rejection). New findings about the mechanism of E- selectin regulation, including data revealing the origin of the activation of p38 MAPK and NF-kappaB by P2 receptors, will define the role of extracellular nucleotides in inflammation and other diseases. Completed studies should offer novel approaches how to control P2 receptors functions that can be useful in the treatment of vascular inflammation, thrombosis, cystic fibrosis and cancer through anti-angiogenic therapies.

由于细胞对各种促炎刺激、组织损伤和细胞死亡的反应，细胞外三磷酸核苷和二磷酸核苷被释放到组织液和血浆中。细胞外核苷酸通过嘌呤能P2型受体对多种细胞发挥作用，引起细胞活化和凋亡。我们的数据显示，细胞外 ATP 诱导内皮细胞 (EC) 活化和 EC 中多种蛋白质的磷酸化，包括粘着斑激酶 (FAK)、桩蛋白、相关粘着斑酪氨酸激酶 (RAFTK)、p130/cas、caveolin-1、Shc 和 p38、SAPK/JNK 和 ERK MAP 激酶。这些蛋白质属于与细胞骨架重排、细胞扩散、运动、细胞增殖和凋亡相关的各种信号转导途径。我们还发现细胞外核苷酸激活核因子 kappa B (NF-kappaB) 并上调 E-选择素的表达，参与白细胞与 EC 的初始相互作用，导致它们迁移到炎症部位。我们打算鉴定负责 E-选择素上调的受体，并阐明 ATP 诱导的信号转导导致 E-选择素基因诱导的机制。获得的结果将有助于阐明涉及嘌呤能受体信号传导的疾病状态的机制（例如炎症、血管生成、动脉粥样硬化、移植物排斥）。关于 E-选择素调节机制的新发现，包括揭示 P2 受体激活 p38 MAPK 和 NF-κB 起源的数据，将定义细胞外核苷酸在炎症和其他疾病中的作用。已完成的研究应该提供如何控制 P2 受体功能的新方法，这些方法可用于通过抗血管生成疗法治疗血管炎症、血栓形成、囊性纤维化和癌症。