Selective induction of endothelial P2Y6 nucleotide receptor promotes vascular inflammation
Selective induction of endothelial P2Y6 nucleotide receptor promotes vascular inflammation
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DOI：
10.1182/blood-2010-10-313957
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发表时间：
2011-02-24
期刊：
影响因子：
20.3
通讯作者：
Eltzschig, Holger K.
中科院分区：
文献类型：
--
作者：
Riegel, Ann-Kathrin;Faigle, Marion;Eltzschig, Holger K.
在系统性炎症反应中，内皮表达的表面分子与协调免疫反应密切相关。先前的研究表明，在急性炎症条件下，细胞外核苷酸释放增强。因此，我们假设内皮核苷酸受体可能在血管炎症中发挥作用。为了验证这一假设，我们进行了筛选实验，将人微血管内皮暴露于炎症刺激下，然后测量P2Y或P2X转录反应。这些研究显示P2Y(6)受体选择性诱导（24小时4倍）。此外，使用实时逆转录聚合酶链反应、Western blot分析或免疫荧光的研究证实，肿瘤坏死因子α或脂多糖（LPS）刺激在体外和体内诱导P2Y(6)具有时间和剂量依赖性。使用MRS 2578作为P2Y(6)受体拮抗剂的研究显示，体外人微血管内皮细胞中核因子κ B报告活性和促炎基因表达减弱。此外，体内药理学或遗传学研究表明，在lps诱导的血管炎症期间，P2Y(6)（-/-）小鼠或P2Y(6)拮抗剂治疗后，炎症反应减弱。这些研究表明P2Y(6)信号在全身性LPS刺激下增强血管炎症中的重要作用，并暗示P2Y(6)受体是全身性炎症反应中的治疗靶点。(血。2011;117 (8):2548 - 2555)
During a systemic inflammatory response endothelial-expressed surface molecules have been strongly implicated in orchestrating immune responses. Previous studies have shown enhanced extracellular nucleotide release during acute inflammatory conditions. Therefore, we hypothesized that endothelial nucleotide receptors could play a role in vascular inflammation. To address this hypothesis, we performed screening experiments and exposed human microvascular endothelia to inflammatory stimuli, followed by measurements of P2Y or P2X transcriptional responses. These studies showed a selective induction of the P2Y(6) receptor (> 4-fold at 24 hours). Moreover, studies that used real-time reverse transcription-polymerase chain reaction, Western blot analysis, or immunofluorescence confirmed time- and dose-dependent induction of P2Y(6) with tumor necrosis factor alpha or Lipopolysaccharide (LPS) stimulation in vitro and in vivo. Studies that used MRS 2578 as P2Y(6) receptor antagonist showed attenuated nuclear factor kappa B reporter activity and proinflammatory gene expression in human microvascular endothelial cells in vitro. Moreover, pharmacologic or genetic in vivo studies showed attenuated inflammatory responses in P2Y(6)(-/-) mice or after P2Y(6) antagonist treatment during LPS-induced vascular inflammation. These studies show an important contribution of P2Y(6) signaling in enhancing vascular inflammation during systemic LPS challenge and implicate the P2Y(6) receptor as a therapeutic target during systemic inflammatory responses. (Blood. 2011;117(8):2548-2555)