DYNAMIC MECHANICAL PERTURBATION AND SIGNAL TRANSDUCTION IN THE AIRWAY EPITHELIUM

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者的研究机构。 据认为，与哮喘发作相关的气道狭窄导致有害机械力直接传递到支气管上皮，并且已经表明这些压缩力增加了促炎和促纤维化介质从培养物中的这些细胞的释放。反复发作的累加效应被认为通过气道壁的间充质细胞诱导气道重塑和胶原沉积，导致在哮喘期间观察到的改变的和不期望的气道特征。 我们已经开发了一种能够向培养中的上皮细胞施加压缩力的装置，该压缩力被认为是在体内气道狭窄期间肺上皮所经历的，以及递送规定的压力振荡的新颖能力，我们认为该压力振荡更准确地模拟了在急性哮喘发作期间递送至气道上皮的力。 为了表征这些效应，将正常人支气管上皮（NHLB）细胞单次或多次暴露于4小时的静态或振荡压力，并测定培养基中的转化生长因子-β 2、内皮素1和2以及纤连蛋白。 还将表征这些蛋白质的基因表达。 然后将NHLB细胞在正常人肺成纤维细胞（NHLF）存在下暴露于先前描述的条件，以表征压缩诱导的细胞损伤期间的上皮-间充质关系。 将评估胶原蛋白（I、III和V）、腱生蛋白和纤连蛋白的基因表达，并将利用多光子激光扫描显微镜对培养物中NHLF细胞的胶原蛋白沉积进行定量。 这项研究将使我们能够检查压缩诱导的细胞损伤对支气管上皮的影响，并将提供深入了解哮喘相关损伤及其参与纤维化发展过程中的上皮-间质关系。 这些研究的最终目的是帮助开发药物干预，以阻止慢性哮喘期间气道重塑和纤维化的进展。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. It is thought that airway narrowing associated with asthmatic episodes results in the transmission of deleterious mechanical forces directly to the bronchial epithelium and it has been shown that these compressive forces augment the release of proinflammatory and profibrogenic mediators from these cells in culture. The additive effect of repeated episodes is thought to induce airway remodeling and collagen deposition by mesenchymal cells of the airway wall resulting in the altered and undesirable airway characteristics seen during asthma. We have developed a device capable of applying compressive forces to epithelial cells in culture thought to be experienced by the pulmonary epithelium during airway narrowing in-vivo, as well as the novel capability of delivering prescribed pressure oscillations that we believe more accurately mimic the forces delivered to the airway epithelium during an acute asthma attack. To characterize these effects, normal human bronchial epithelial (NHLB) cells will be subjected to a single or multiple exposure to 4 hours of static or oscillatory pressures and the media will be assayed for transforming growth factor-¿¿2, endothelin 1 and 2, and fibronectin. Gene expression of these proteins will also be characterized. NHLB cells will then be exposed to the conditions described earlier in the presence of normal human lung fibroblasts (NHLF) to characterize the epithelial-mesenchymal relationship during compression induced cell injury. Gene expression of collagen (I, III and V), tenascin and fibronectin will be assessed and multiphoton laser scanning microscopy will be utilized to quantify collagen deposition by NHLF cells in culture. This study will allow us to examine the effects of compression induced cell injury on the bronchial epithelium and will provide insight into the epithelial-mesenchymal relationship during asthma related injury and its involvement in the development of fibrosis. The ultimate goal of these studies is to aid in the development of pharmacologic interventions to impede the progression of airway remodeling and fibrosis during chronic asthma.