DYNAMIC MECHANICAL PERTURBATION AND SIGNAL TRANSDUCTION IN THE AIRWAY EPITHELIUM

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 与哮喘发作相关的气道狭窄被认为是有害的机械力直接传递到支气管上皮的结果，已有研究表明，这些压力会在培养中增加这些细胞释放促炎和促纤维化介质。反复发作的相加效应被认为是诱导气道重塑和气道壁间充质细胞的胶原沉积，从而导致哮喘期间出现的改变和不良的气道特征。我们已经开发出一种设备，能够对培养的上皮细胞施加压缩力，被认为是在体内气道狭窄期间肺上皮细胞所经历的，以及提供规定的压力振荡的新能力，我们相信这种能力更准确地模拟了在急性哮喘发作期间传递到气道上皮细胞的力。为了表征这些效应，正常的人支气管上皮(NHLB)细胞将受到一次或多次4小时的静态或振荡压力，并将检测培养液中转化生长因子-2、内皮素1和2以及纤维连接蛋白的含量。这些蛋白质的基因表达也将被表征。然后，在正常人肺成纤维细胞(NHLF)存在的情况下，NHLB细胞将暴露在前面描述的条件下，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(I、III和V)、肌腱蛋白和纤维连接蛋白的基因表达，并将利用多光子激光扫描显微镜定量测定NHLF细胞在培养中的胶原沉积。这项研究将使我们能够检查压迫诱导的细胞损伤对支气管上皮的影响，并将为深入了解哮喘相关损伤中的上皮-间充质关系及其在纤维化发展中的作用提供依据。这些研究的最终目标是帮助开发药物干预措施，以阻止慢性哮喘期间呼吸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.