PULMONARY ENDOTHELIAL CELL TRANSCYTOSIS AND ACUTE LUNG INJURY AFTER HEMORRHAGIC

肺内皮细胞转胞作用和出血后急性肺损伤

• 批准号: 6861600
• 负责人: Bruce Robert Pitt
• 金额: $ 21.35万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-07-01 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6861600
• 关键词: biological signal transduction; confocal scanning microscopy; fluorescence resonance energy transfer; genetically modified animals; hemorrhagic shock; inflammation; injury; laboratory mouse; laboratory rat; lung; muscle cells; mutant; myeloperoxidase; nitric oxide; respiratory epithelium; resuscitation; serum albumin; smooth muscle; stress; tissue /cell culture; transcytosis; trauma

Pulmonary endothelium is an early locus of functional and structural changes that contribute to the genesis and/or maintenance of acute lung injury(ALl). We will define the mechanisms of transport and the vascular actions of key inflammatory-modulatory macromolecules [e.g. S-nitroso-albumin (SNO-AIb) and myeloperoxidase (MPO)] that are elevated in the circulation after resuscitation from hemorrhagic shock (HS). Accordingly specific aims are to: Aim #1. Define the physiological mechanism of transcytosis of SNO-AIb in cultured pulmonary endothelium. We will use real time multimode imaging (confocal and multiphoton scanning laser microscopy and total internal reflection fluorescence microscopy, FRET) to define molecular events associated with vascular cell binding, transcytosis and SNO-AIb mediatied signaling via S-nitrosation and/or activation of guanylyl cyclase. Aim #2. Reveal the physiological mechanisms of MPO transcytosis in live intact pulmonary endothelium Aim #3. Explore the biochemical mechanisms by which MPO affects SNO-albumin signaling in pulmonary vascular cells and intact lung. To reveal the central pathways for intra- and extracellular regulatory MPO-dependent modulation of vascular NO signaling, we will examine the actions of MPO on SNO-albumin transfer and decomposition kinetics in increasingly integrated biological systems including: a) cell free models in vitro; b) rat lung microvacular endothelial cells (RLMVEC); and c) rat lung microvasccular smooth muscle cell and RLMVEC co-culture system. Specific Aim #4. Place the pathophysiological contributions of SNO-albumin transport, redox status and MPO into the context of ALl after resuscitation from HS. We will contrast the extent of ALl (permeability index, neutrophil accumulation, morphological changes,pulmonary vasoregulation) in isolated perfused lungs and intact mice of wildtype and caveolin-1 and MPO homozygous null mutants.

肺内皮是功能和结构变化的早期位点，其有助于急性肺损伤(AL1)的发生和/或维持。我们将定义关键炎症调节大分子的运输机制和血管作用[例如失血性休克 (HS) 复苏后循环中的 S-亚硝基白蛋白 (SNO-AIb) 和髓过氧化物酶 (MPO)] 升高。因此，具体目标是： 目标#1。定义培养肺内皮细胞中 SNO-AIb 转胞吞作用的生理机制。我们将使用实时多模式成像（共焦和多光子扫描激光显微镜和全内反射荧光显微镜，FRET）来定义与血管细胞结合、转胞吞作用和通过 S-亚硝化和/或鸟苷基激活介导的 SNO-AIb 介导的信号传导相关的分子事件环化酶。目标#2。揭示活体完整肺内皮中 MPO 转胞吞作用的生理机制目标#3。探索 MPO 影响肺血管细胞和完整肺中 SNO-白蛋白信号传导的生化机制。揭示细胞内和细胞外 MPO 依赖性调节的中心途径 血管 NO 信号传导的调节，我们将检查 MPO 对 SNO-白蛋白的作用 日益一体化的生物系统中的转移和分解动力学，包括：a) 体外无细胞模型； b)大鼠肺微血管内皮细胞(RLMVEC)； c)大鼠肺微血管平滑肌细胞和RLMVEC共培养系统。具体目标#4。将 SNO-白蛋白转运、氧化还原状态和 MPO 的病理生理学贡献置于 HS 复苏后 AL1 的背景中。我们将对比 ALl 的程度（渗透性 野生型和 Caveolin-1 和 MPO 纯合无效突变体的离体灌注肺和完整小鼠中的指数、中性粒细胞积累、形态变化、肺血管调节）。