Regulation of platelet responses triggered by Toll-like receptor 2 and 4 ligands is another non-genomic role of nuclear factor-kappaB
Regulation of platelet responses triggered by Toll-like receptor 2 and 4 ligands is another non-genomic role of nuclear factor-kappaB
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DOI：
10.1016/j.thromres.2013.11.028
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发表时间：
2014-02-01
影响因子：
7.5
通讯作者：
Schattner, Mirta
中科院分区：
文献类型：
--
作者：
Rivadeneyra, Leonardo;Carestia, Agostina;Schattner, Mirta
简介：血小板表达 Toll 样受体 (TLR)，可识别病原体的分子成分，并在有核细胞中通过核因子 kappaB (NF-kappa B) 激活引发免疫反应。我们已经证明 NF-kappa B 介导血小板活化以响应经典激动剂，这表明该转录因子在血小板中发挥非基因组功能。本研究的目的是确定 NF-kappa B 激活是否是参与 TLR2 和 4 介导的血小板反应的下游信号。材料和方法：用 Lumi 聚集计测量聚集和 ATP 释放。通过细胞计数法测量纤维蛋白原结合、P-选择素和 CD40 配体 (CD40L) 水平以及血小板-中性粒细胞聚集体。通过蛋白质印迹法测定 I kappa B α (I kappa B α) 降解和 p65 磷酸化，通过 ELISA 测定冯维勒布兰德因子 (vWF)。 结果：用 Pam3CSK4 或 LPS 刺激血小板导致 I kappa B α 降解和 p65 磷酸化。这些反应被 TLR2 和 4 阻断所抑制，并被凝血酶协同作用。 Pam3CSK4 触发聚集、纤维蛋白原结合以及 ATP 和 vWF 释放。除了 vWF 释放外，LPS 本身并不诱导血小板反应，但它确实增强了凝血酶诱导的聚集、纤维蛋白原结合和 ATP 分泌。 Pam3CSK4（而非 LPS）诱导 P-选择素和 CD40L 表达以及混合聚集体形成。除 CD40L 表达外，所有这些反应在用 NF-κ B 抑制剂 BAY 11-7082 或 Ro 106-9920 处理的血小板中均受到抑制。结论：TLR2 和 4 激动剂通过 NF-κ B 触发血小板活化反应。这些数据显示了 NF-κ B 在血小板中的另一种非基因组功能，并强调该分子作为预防炎症或传染病中血小板活化的潜在靶点。 (C) 2013 Elsevier Ltd. 保留所有权利。
Introduction: Platelets express Toll-like receptors (TLRs) that recognise molecular components of pathogens and, in nucleated cells, elicit immune responses through nuclear factor-kappaB (NF-kappa B) activation. We have shown that NF-kappa B mediates platelet activation in response to classical agonists, suggesting that this transcription factor exerts non-genomic functions in platelets. The aim of this study was to determine whether NF-kappa B activation is a downstream signal involved in TLR2 and 4-mediated platelet responses.Material and methods: Aggregation and ATP release were measured with a Lumi-aggregometer. Fibrinogen binding, P-selectin and CD40 ligand (CD40L) levels and platelet-neutrophil aggregates were measured by cytometry. I kappa B alpha (I kappa B alpha) degradation and p65 phosphorylation were determined by Western blot and von Willebrand factor (vWF) by ELISA.Results: Platelet stimulation with Pam3CSK4 or LPS resulted in I kappa B alpha degradation and p65 phosphorylation. These responses were suppressed by TLR2 and 4 blocking and synergised by thrombin. Aggregation, fibrinogen binding and ATP and vWF release were triggered by Pam3CSK4. LPS did not induce platelet responses per se, except for vWF release, but it did potentiate thrombin-induced aggregation, fibrinogen binding and ATP secretion. Pam3CSK4, but not LPS, induced P-selectin and CD40L expression and mixed aggregate formation. All of these responses, except for CD40L expression, were inhibited in platelets treated with the NF-kappa B inhibitors BAY 11-7082 or Ro 106-9920.Conclusion: TLR2 and 4 agonists trigger platelet activation responses through NF-kappa B. These data show another non-genomic function of NF-kappa B in platelets and highlight this molecule as a potential target to prevent platelet activation in inflammatory or infectious diseases. (C) 2013 Elsevier Ltd. All rights reserved.