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.

肺部内皮是功能和结构变化的早期基因座，有助于急性肺损伤的起源和/或维持（全部）。我们将定义运输机制和关键炎症调节大分子的血管作用[例如S-硝基 - α-α（SNO-AIB）和髓过氧化物酶（MPO）]，在出血性休克（HS）复苏后循环中升高。因此，具体目的是： 目标＃1。定义培养的肺部内皮中SNO-AIB转胞细胞增多症的生理机制。我们将使用实时多模成像（共聚焦和多光子扫描激光显微镜以及总内反射荧光显微镜，FRET）来定义与血管细胞结合，跨念珠菌结合，SNO-AIB通过S-硝化信号传导和/ORISITATION和/ORIPITATION GUANYLELLLELL CYCLase的分子事件。目标＃2。揭示了活完整的肺部内皮目标中MPO转胞膜的生理机制＃3。探索MPO影响肺血管细胞和完整肺中SNO-Albumin信号传导的生化机制。揭示细胞内和细胞外MPO依赖性的中心途径 调节血管无信号传导，我们将检查MPO对SNO-Albumin的作用 在日益整合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）大鼠肺微分流内皮细胞（RLMVEC）； c）大鼠肺微伏平滑肌细胞和RLMVEC共培养系统。特定目标＃4。将SNO-Albumin转运，氧化还原状态和MPO的病理生理贡献置于HS复苏后的所有情况下。我们将对比所有的程度（渗透性 在孤立的灌注肺和野生型和小窝蛋白-1和MPO纯合无效突变体中，索引，中性粒细胞的积累，形态变化，肺血管调节）。