Intact Toll-like receptor 9 signaling in neutrophils modulates normal thrombogenesis in mice
Intact Toll-like receptor 9 signaling in neutrophils modulates normal thrombogenesis in mice
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DOI：
10.1016/j.jvs.2015.08.070
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发表时间：
2016-11-01
影响因子：
4.3
通讯作者：
Henke, Peter K.
中科院分区：
文献类型：
--
作者：
El-Sayed, Osama M.;Dewyer, Nicholas A.;Henke, Peter K.
背景：toll样受体9 （Tlr9）信号在无菌炎症过程中起重要作用，其缺失可导致小鼠静脉血栓形成（VT）消退受损。本研究的目的是确定Tlr9的缺失是否影响VT无菌坏死、凋亡和中性粒细胞胞外陷阱（NET）的产生。方法：采用野生型（WT）和Tlr9（-/-）小鼠的VT停滞和非停滞小鼠模型，评估血栓大小，测定NETs、坏死和凋亡标志物。使用抗多形核中性粒细胞（PMN）和抗血小板抗体策略来确定其在WT和Tlr9（-/-）小鼠中的细胞作用及其作用。结果：第2天，与WT小鼠相比，Tlr9（-/-）小鼠的瘀血血栓增大62% (n = 6-10)，尿酸水平增加1.4倍，凋亡细胞增加1.7倍，瓜氨酸组蛋白增加2倍，肽精氨酸脱亚胺酶4 （PAD4）增加2倍，弹性酶增加1.5倍，组织因子途径抑制剂减少2.4倍（均n = 4-7; P < 0.05）。相反，Tlr9（-/-）小鼠（n = 4-6）的非停滞性血栓大小没有显著差异，它们没有坏死或NET标记物升高。在接受脱氧核糖核酸酶I治疗的WT或Tlr9（-/-）小鼠，以及无法形成NETs的PAD4（-/-）小鼠，在2天时间点上，瘀血血栓大小没有减少。在Tlr9（-/-）小鼠中，经过PMN消耗（n = 8-10），停滞血栓大小减少18%，瓜氨酸组蛋白减少29倍，弹性蛋白酶减少1.3倍，组织因子途径抑制剂增加1.5倍（均n = 6; P < 0.05）。最后，血小板消耗（>减少90%）并没有显著减少Tlr9（-/-）小鼠的瘀血血栓大小。结论：这些数据表明，血栓形成模型影响Tlr9的血栓形成机制，功能Tlr9信号在PMNs中，而不是在血小板或NETs中，是早期停滞实验性静脉血栓形成的重要机制。
Background: Deletion of Toll-like receptor 9 (Tlr9) signaling, which is important for sterile inflammatory processes, results in impaired resolution of venous thrombosis (VT) in mice. The purpose of this study was to determine if deletion of Tlr9 affected sterile necrosis, apoptosis, and neutrophil extracellular trap (NET) production in VT.Methods: Stasis and nonstasis murine models of VT were used in wild-type (WT) and Tlr9(-/-) mice, with assessment of thrombus size and determination of NETs, necrosis, and apoptosis markers. Anti-polymorphonuclear neutrophil (PMN) and antiplatelet antibody strategies were used to determine the cellular roles and their roles in WT and Tlr9(-/-) mice.Results: At 2 days, stasis thrombi in Tlr9(-/-) mice were 62% larger (n = 6-10), with 1.4-fold increased uric acid levels, 1.7-fold more apoptotic cells, 2-fold increased citrullinated histones, 2-fold increased peptidylarginine deiminase 4 (PAD4), and 1.5-fold increased elastase and a 2.4-fold reduction in tissue factor pathway inhibitor compared with WT mice (all n = 4-7; P < .05). In contrast, the sizes of nonstasis thrombi were not significantly different in Tlr9(-/-) mice (n = 4-6), and they did not have elevated necrosis or NET markers. Stasis thrombus size was not reduced at the 2-day time point in WT or Tlr9(-/-) mice that received treatment with deoxyribonuclease I or in PAD4(-/-) mice, which are incapable of forming NETs. In Tlr9(-/-) mice undergoing PMN depletion (n = 8-10), stasis thrombus size was reduced 18% and was associated with 29-fold decreased citrullinated histones, 1.3-fold decreased elastase, and 1.5-fold increased tissue factor pathway inhibitor (all n = 6; P < .05). Last, platelet depletion (>90% reduction) did not significantly reduce stasis thrombus size in Tlr9(-/-) mice.Conclusions: These data suggest that the thrombogenic model affects Tlr9 thrombogenic mechanisms and that functional Tlr9 signaling in PMNs, but not in platelets or NETs, is an important mechanism in early stasis experimental venous thrombogenesis.