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TLR2 in Sepsis-Induced Coagulopathy, Endothelial Leak, and Pulmonary Dysfunction

TLR2 在脓毒症引起的凝血病、内皮渗漏和肺功能障碍中的作用

基本信息

批准号：
7860482
负责人：
Judith Hellman
金额：
$ 34.52万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-06-01 至 2012-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7860482
关键词：
Acute Lung Injury Adherence Agonist Albumins Anticoagulants Anticoagulation Antithrombin III Bacterial Counts Blood Blood Coagulation Disorders Blood Platelets Blood Vessels Blood coagulation Coagulation Process Complex Data Edema Endothelial Cells Endothelium Family Fibrin fragment D Fibrinogen Fibrinolysis Fibrinolytic Agents Functional disorder Gram-Negative Bacteria Histologic Human Hypoxia Immune response In Vitro Infection Inflammation Inflammatory Knock-out Knockout Mice Life Ligands Lipoproteins Lung Measures Mediating Mediator of activation protein Modeling Mus Nitric Oxide Nitrites Organ failure Pathway interactions Peritonitis Permeability Plasma Plasminogen Activator Inhibitor 1 Pneumonia Process Protein C Pulmonary Edema Receptor Activation Relative (related person)Respiratory Failure Respiratory physiology Role Sepsis Shock Signal Pathway System TFPI Testing Thromboplastin Thrombosis Time Toll-Like Receptor 2 Toll-like receptors Toxic effect Vascular Permeabilities Weight Wild Type Mouse in vivo inflammatory marker inhibitor/antagonist insight microbial microorganism monolayer mortality neutrophil respiratory response treatment effect vasoconstriction wet lung

项目摘要

The central hypothesis is that activation of Toll-like receptor (TLR) 2 contributes to the connected processes of endothelial dysfunction, coagulopathy, and increased vascular permeability in sepsis. TLR2 mediates the inflammatory effects of bacterial lipoproteins. The studies will define mechanisms by which TLR2 agonists modulate coagulation pathways in endothelial cells, and will assess the functional significance of TLR2 activation on endothelial permeability and coagulopathy in sepsis. These studies will provide insights into the mechanisms of coagulopathy, vascular leak, and respiratory dysfunction in sepsis. TLR2 agonists are present in all of the major classes of microorganisms that cause sepsis. Thus if TLR2 is important in sepsis-induced coagulopathy, vascular leak or respiratory failure, then TLR2 signaling pathways could be suitable targets for sepsis therapies. Specific Aim #1: Define mechanisms by which TLR2 activation modulates endothelial cell (EC) expression of coagulation pathway factors in vitro. Studies will test the hypotheses that TLR2 agonists alter expression of factors involved in coagulation, anticoagulation, and fibrinolysis: 1) through NF-B, and 2) through additional mediators, including TGF-, TNF, and/or NO. EC will be treated with TLR2 agonists, and expression of tissue factor (TF), tissue factor pathway inhibitor (TFPI), and plasminogen activator inhibitor type 1 (PAI-1) will be quantified. Mechanisms by which TLR2 agonists modulate coagulation pathways will be defined using EC from knockout mice, and using targeted inhibitors with human endothelial cells. Specific Aim #2: Assess the effects of TLR2 activation on endothelial permeability in vitro. Studies will test the hypotheses that TLR2 activation increases endothelial leakiness, as assessed by permeability of EC monolayers to albumin. Specific Aim #3: Define the functional significance of TLR2 activation on coagulopathy and on lung vascular permeability in sepsis. Sepsis will be induced in mice using peritonitis and pneumonia models. Studies will compare responses of TLR2 knockout mice with those of wild-type mice, and will assess the relative importance of TLR2 in the pathophysiology of Grampositive versus Gram-negative sepsis. Blood coagulation times will be measured, and levels of factors involved in coagulation and fibrinolysis will be quantified in blood and in lung. Lung vascular leakiness will be assessed using lung wet:dry weight ratios and permeability to albumin. Histologic analyses will assess for microvascular thrombosis, architectural changes, evidence of pulmonary edema, and lung expression of PAI-1 and TF.

中心假设是Toll样受体（TLR）2的激活有助于脓毒症中内皮功能障碍、凝血障碍和血管通透性增加的相关过程。TLR 2介导细菌脂蛋白的炎症作用。这些研究将确定TLR 2激动剂调节内皮细胞凝血途径的机制，并将评估TLR 2活化对脓毒症中内皮通透性和凝血障碍的功能意义。这些研究将为脓毒症中凝血功能障碍、血管渗漏和呼吸功能障碍的机制提供见解。TLR 2激动剂存在于引起脓毒症的所有主要微生物类别中。因此，如果TLR 2在脓毒症诱导的凝血病、血管渗漏或呼吸衰竭中是重要的，那么TLR 2信号传导途径可能是脓毒症治疗的合适靶点。具体目标#1：定义TLR 2激活在体外调节凝血途径因子的内皮细胞（EC）表达的机制。研究将检验TLR 2激动剂改变凝血、抗凝和纤溶相关因子表达的假设：1）通过NF-B，和2）通过另外的介质，包括TGF-β、TNF和/或NO。EC将用TLR 2激动剂和组织因子（TF）、组织因子途径抑制剂（TFPI）的表达治疗，和纤溶酶原激活物抑制剂1型（派-1）将被定量。TLR 2激动剂调节凝血途径的机制将使用来自敲除小鼠的EC和使用靶向抑制剂与人内皮细胞来定义。具体目标#2：评估TLR 2激活对体外内皮渗透性的影响。研究将检验TLR 2激活增加内皮渗漏的假设，如通过EC单层对白蛋白的渗透性所评估的。具体目标#3：确定TLR 2活化对脓毒症凝血障碍和肺血管通透性的功能意义。将使用腹膜炎和肺炎模型在小鼠中诱导脓毒症。研究将比较TLR 2敲除小鼠与野生型小鼠的应答，并将评估TLR 2在革兰氏阳性与革兰氏阴性脓毒症的病理生理学中的相对重要性。将测量血液凝固时间，并定量血液和肺中凝血和纤维蛋白溶解相关因子的水平。将使用肺湿重：干重比和白蛋白渗透性评估肺血管渗漏。组织学分析将评估微血管血栓形成、结构变化、肺水肿证据以及派-1和TF的肺表达。