S1P-fluid therapy to reduce hemorrhagic shock & intoxication-induced injury

S1P 液体疗法可减少失血性休克

• 批准号: 9310336
• 负责人: JEROME W BRESLIN
• 金额: $ 28.78万
• 依托单位: UNIVERSITY OF SOUTH FLORIDA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-15 至 2020-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9310336
• 关键词: Acute; Adult; Agonist; Alcohol-Induced Disorders; Alcoholic Intoxication; Alcohols; Apoptotic; Architecture; Biochemical; Biological Preservation; Blood Pressure; Cell Survival; Clinical; Complement; Complex; Confocal Microscopy; Conscious; Data; Development; Dose; Edema; Effectiveness; Endothelial Cells; Ethanol Metabolism; Extravasation; Fluid Therapy; Foundations; Functional disorder; Goals; Health Care Costs; Hemorrhage; Hemorrhagic Shock; Hypotension; Individual; Inflammation; Inflammatory; Injury; Innovative Therapy; Intercellular Junctions; Intervention; Intoxication; Investigation; Knowledge; Laboratory Finding; Lasers; Life; Lipids; Liquid substance; Mediating; Mesentery; Microvascular Permeability; Mission; Modeling; Outcome; Pathway interactions; Patients; Pharmacology; Physiological; Protocols documentation; Public Health; Rattus; Research; Resuscitation; Rodent Model; Role; Severities; Shock; Signal Transduction; Small Interfering RNA; Sphingosine-1-Phosphate Receptor; System; Testing; Therapeutic; Time; Trauma; Trauma patient; Traumatic injury; United States National Institutes of Health; Work; alcohol exposure; binge drinking; design; disability; effective therapy; improved; in vivo; in vivo Model; innovation; intravenous administration; intravital microscopy; knock-down; mimetics; monolayer; postcapillary venule; pre-clinical; prevent; receptor; repaired; sphingosine 1-phosphate; success; translational impact; trend

There is a fundamental gap in understanding pertaining to why standard fluid resuscitation protocols work so poorly in alcohol-intoxicated trauma victims with hemorrhagic shock, representing a significant portion of all trauma cases. Continued existence of this gap represents an important problem, because until it is filled, the development of effective therapies to prevent systemic inflammation and prolonged hypotension in trauma patients will continue to be unsuccessful. The long-term goal of this study is to develop effective therapies to reduce elevated microvascular permeability and edema. The overall objective in this application is to determine the effectiveness of the bioactive lipid sphingosine-1-phosphate (S1P) for ameliorating alcohol-induced elevated microvascular permeability. The central hypothesis is that S1P reduces alcohol/injury-induced microvascular hyperpermeability by activating S1P receptor-1 (S1P1R), causing initial junctional repair, and longer term pro-survival signals, thereby promoting a healthy endothelial barrier. The rationale for the proposed research is that it will provide important details obtained from a relevant model of combined alcohol intoxication and hemorrhagic injury that will be needed for development of effective fluid resuscitation strategies. Guided by strong preliminary data, the central hypothesis will be tested with three specific aims: Aim 1 is to determine the optimal dose and window of opportunity for S1P therapy after combined alcohol intoxication and hemorrhagic shock. Aim 2 is to test selective S1P1R agonist therapies to ameliorate alcohol/shock-induced microvascular hyperpermeability and hypotension. Aim 3 is to determine the extent to which S1P-induced, RhoA-mediated junctional enhancement and cell survival signals contribute to ameliorate endothelial barrier dysfunction. These aims will utilize an established conscious rat model of combined alcohol intoxication and hemorrhagic shock and resuscitation. The experimental endpoints will be the extent to which S1P or S1P1R agonists ameliorate hypotension, microvascular leakage in the gut, and gut barrier injury. The in vivo studies will be complemented by an investigation of the ability of S1P or S1P1R agonists to improve barrier integrity of cultured microvascular endothelial cell monolayers exposed to alcohol. Pharmacologic antagonists or specific siRNA knockdown will be used to assess the role of the S1P1R. A combination of intravital microscopy, laser confocal microscopy, and biochemical analyses will be used to study microvascular endothelial junctional architecture and survival signals in individual endothelial cells of postcapillary venules. The contribution of the proposed research will be significant because it will provide proof of concept from a rodent model that alcohol-induced amplifications of elevated microvascular permeability contribute to the lack of success with current clinical interventions used to resuscitate alcohol-intoxicated hemorrhagic shock patients. The proposed research is innovative because it represents a substantive departure from the status quo by investigating a therapeutic approach to ameliorate alcohol intoxication-induced microvascular hyperpermeability as the key approach to stabilize blood pressure.

理解与标准流体复苏方案为何有效的理解有关的根本差距 患有出血性休克的酒精感染创伤受害者的不良 创伤病例。持续存在此差距是一个重要的问题，因为直到填充为止 开发有效的疗法，以防止全身炎症和创伤长期低血压 患者将继续失败。这项研究的长期目标是开发有效的疗法 减少升高的微血管通透性和水肿。该应用程序的总体目的是确定 生物活性脂质鞘氨醇1-磷酸（S1P）的有效性用于改善酒精诱导的 微血管渗透率升高。中心假设是S1P降低了酒精/损伤诱导的 通过激活S1P受体1（S1P1R），导致初始连接修复和 长期的亲生信号，从而促进了健康的内皮屏障。提议的理由 研究是，它将提供从合并酒精中毒的相关模型获得的重要细节 以及开发有效的液体复苏策略所需的出血损伤。指导 强大的初步数据，中央假设将通过三个具体目标进行测试：目标1是确定 酒精中毒和出血后S1P治疗的最佳剂量和机会窗口 震惊。 AIM 2是测试选择性S1P1R激动剂疗法以改善酒精/休克诱导的微血管 过度性和低血压。 AIM 3是确定S1P诱导的RhoA介导的程度 连接增强和细胞存活信号有助于改善内皮屏障功能障碍。这些 AIMS将利用既定的意识大鼠模型，用于合并的酒精中毒和出血性休克 和复苏。实验终点将是S1P或S1P1R激动剂改善的程度 低血压，肠道中的微血管泄漏以及肠道屏障损伤。体内研究将得到补充 通过研究S1P或S1P1R激动剂改善培养微血管屏障完整性的能力 内皮细胞单层暴露于酒精。药理学拮抗剂或特定的siRNA敲低将 用于评估S1P1R的作用。插入显微镜，激光共聚焦显微镜的组合， 生化分析将用于研究微血管内皮结构和生存 后毛细血管静脉的单个内皮细胞中的信号。拟议研究的贡献将是 重要的是因为它将提供啮齿动物模型的概念证明 升高的微血管通透性升高导致缺乏成功的临床干预措施的成功 恢复酗酒的出血性休克患者。拟议的研究具有创新性，因为它 通过调查改善治疗方法来代表与现状的实质性不同 酒精中毒引起的微血管高温性是稳定血压的关键方法。