Hemostasis System in Acute Inflammation-Sepsis

急性炎症败血症的止血系统

基本信息

批准号：
7406633
负责人：
FRANCIS J CASTELLINO
金额：
$ 51.6万
依托单位：
UNIVERSITY OF NOTRE DAME
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/7406633
关键词：
Acute Adhesions Adverse effects Agreement Animal Model Animals Anti-Inflammatory Agents Anti-inflammatory Anticoagulants Anticoagulation Antiplasmin Antithrombin III Appearance Area Attenuated Binding Biological Assay Blood Chemical Analysis Blood Coagulation Disorders Blood Platelets Blood Pressure Blood coagulation CCL2 gene Cardiovascular Diseases Cell Adhesion Cell Adhesion Molecules Cell Communication Cells Chemicals Clinical Coagulation Process Complex Condition Data Deposition Development Diagnosis Disease Disease Progression Disseminated Intravascular Coagulation Dose Down-Regulation Endopeptidases Endothelial Cells Endothelium Endotoxemia Endotoxins Environment Enzyme-Linked Immunosorbent Assay Enzymes Equilibrium Evaluation Event Evolution Extravasation F Factor Fibrin Fibrinogen Receptors Fibrinolysis Gene Expression Gene Mutation Gene Proteins Gene Targeting Generations Genes Glycine decarboxylase Goals Heart Heart Rate Hemostatic function Histocompatibility Testing Individual Inflammation Inflammation Mediators Inflammatory Inflammatory Response Infusion procedures Injection of therapeutic agent Intervention Kidney Kinins Knowledge Lead Leukocyte Rolling Leukocytes Lipopolysaccharides Liquid substance Liver Lung Lymphokines Measures Mediating Mediator of activation protein Messenger RNA Methods Modeling Monitor Morbidity - disease rate Multiple Organ Failure Mus Mutation Nature Organ Organ failure Pathology Pathway interactions Patients Peptide Hydrolases Phenotype Plasma Plasminogen Plasminogen Activator Plasminogen Activator Inhibitor 1 Process Production Property Proteins Pump Regulation Relative (related person)Reverse Transcriptase Polymerase Chain Reaction Risk Factors Role SELP gene SM 22 muscle protein Selectins Sepsis Septic Shock Shock Signal Pathway Signal Transduction Slice Staging Surface System Systemic infection TFPI Testing Therapeutic Intervention Thrombin Thrombomodulin Thromboplastin Thrombosis Time Tissues Trees Up-Regulation Urokinase Venous Wild Type Mouse activated Protein C animal tissue attenuation base cell injury chemokine cytokine disease phenotype functional disability in vivo inhibitor/antagonist insight knowledge base macrophage mortality mouse model neutrophil novel strategies progesterone 11-hemisuccinate-(2-iodohistamine)programs receptor research study response septic tool

项目摘要

There is a strong relationship between inflammation and hemostasis that is based on the understanding that a proinflammatory environment is also procoagulant. Events that lead to the generation of the Factor (F) Vlla/Tissue Factor (TF) complex in an acute inflammatory state, such as gram-negative sepsis, begin with endotoxin-mediated activation of ieukocytes and endothelial cells, and cytokine and chemokine upregulation from these altered cells, along with expression of TF. The consequent generation of the procoagulants, thrombin and FXa, initiates signaling pathways via interactions of these proteases with protease activated cellular receptors, and thus mediate further cellular (e.g., platelet, endothelial cell, leukocyte) responses that are involved in blood coagulation, as well as inflammation and vessel formation. The activation of these cells also results in the expression of adhesion molecules on their surfaces, thereby facilitating leukocyte binding to the endothelium. This interaction is the first step in ultimate extravasation of neutrophils and macrophages into organs, thereby causing severe organ damage in the advanced septic state. Thus, it is our basic hypothesis that attenuation of inflammation may result from inhibition of coagulation and vice versa, and sepsis models using appropriate gene-targeted mice will allow an understanding of these relationships in vivo at the level of the gene. The overall goal of this proposal is to dissect the in vivo relationships between individual genes of hemostasis and inflammation that occur in an acute model of the serious inflammatory disease, gram-negative sepsis, with its progression to severe sepsis and septic shock. Mice with single and combined genetic alterations in the pathways of hemostasis will be employed with endotoxin (LPS)-mediated models of sepsis in order to monitor the relationships between hemostasis and inflammation in the progression of the disease. Specifically, 5 highly interconnected specific aims are proposed: (1) to employ mice (and isolated endothelial cells and adherent macrophages) with genetic alterations of specific hemostasis-related genes in examining the progression of induced sepsis to severe sepsis and septic shock, and ultimate survival; (2) to assess temporal responses, after injection of LPS, of the systemic coagulation, anticoagulation, and fibrinolytic systems in these genotypically-distinct mice; (3) to measure in these same mice, the temporal plasma and organ responses of specific cytokines, chemokines, soluble adhesion proteins, and other inflammatory mediators; (4) to determine the temporal nature of organ and cell damage in tissue slices in these mice after LPS administration; and (5) to employ additional mice with combined deficiencies of hemostasis- and inflammation-related genes to further understand the in vivo mechanisms involved in their sepsis-related effects. It is expected that the results of this study will allow an in vivo evaluation of the roles of specific hemostasis- and inflammation-related genes in the development and course of this model of acute inflammatory disease, and will provide groundwork for therapeutic interventions to attenuate the morbidity and mortality associated with its downward progression.

炎症和止血之间存在密切关系，这是基于促炎环境也是促凝血的这一认识。导致急性炎症状态（例如革兰氏阴性脓毒症）产生因子（F）VIIa/组织因子（TF）复合物的事件始于内毒素介导的白细胞和内皮细胞的激活以及细胞因子和趋化因子的上调来自这些改变的细胞，以及 TF 的表达。随后产生促凝血剂、凝血酶和 FXa，通过这些蛋白酶与蛋白酶激活的细胞受体的相互作用启动信号传导途径，从而介导参与血液凝固的进一步细胞（例如血小板、内皮细胞、白细胞）反应如炎症和血管形成。这些细胞的激活也会导致其表面粘附分子的表达，从而促进白细胞与内皮细胞的结合。这种相互作用是中性粒细胞和巨噬细胞最终外渗到器官中的第一步，从而在晚期脓毒症状态下造成严重的器官损伤。因此，我们的基本假设是炎症的减弱可能是由于凝血的抑制引起的，反之亦然，并且使用适当的基因靶向小鼠的脓毒症模型将允许在基因水平上了解这些体内关系。该提案的总体目标是剖析严重炎症性疾病、革兰氏阴性脓毒症的急性模型中发生的止血和炎症的各个基因之间的体内关系，及其进展为严重脓毒症和脓毒性休克。止血途径中具有单一和组合遗传改变的小鼠将被用于内毒素（LPS）介导的脓毒症模型，以监测疾病进展中止血和炎症之间的关系。具体而言，提出了 5 个高度相互关联的具体目标：（1）使用具有特定止血相关基因遗传改变的小鼠（以及分离的内皮细胞和贴壁巨噬细胞）检查诱发脓毒症向严重脓毒症和脓毒性休克的进展以及最终生存率； (2) 评估注射 LPS 后，这些基因型不同的小鼠的全身凝血、抗凝和纤溶系统的时间反应； (3) 在这些相同的小鼠中测量特定细胞因子、趋化因子、可溶性粘附蛋白和其他炎症介质的颞叶血浆和器官反应； (4) 测定LPS给药后这些小鼠组织切片中器官和细胞损伤的时间性质； (5) 使用其他具有止血和炎症相关基因联合缺陷的小鼠来进一步了解其脓毒症相关效应所涉及的体内机制。预计这项研究的结果将能够对特定止血和炎症相关基因在该模型的发展和过程中的作用进行体内评估。急性炎症性疾病，并将为治疗干预措施奠定基础，以减轻与其下降进展相关的发病率和死亡率。