Surfactant Proteins and NO in Inflammatory Disease

描述(由申请人提供)：本研究项目的主要目标是调查化学物质如何在肺部炎症疾病的启动和传播中发挥作用。特别是，我们将在一个肺部炎症的动物模型中研究改变NO化学的影响，即消融SP-D基因。在没有感染源的情况下，缺乏SP-D基因的小鼠会在肺内发展出一种非特异性炎症疾病。本病以肺实质浸润性病变、中性粒细胞脂蛋白沉积症和肺组织NO化学改变(组织内S亚硝硫醇丢失，硝基酪氨酸增多，细支气管肺泡灌洗液中硝酸盐相对增多)为特征。因此，该动物模型为研究NO在肺部炎症中的作用提供了一个理想的机会。我们将在这个模型中使用药物和遗传干预来控制NO的产生，并检查这些控制对疾病发展的影响。此外，我们认为正常信号NO的丢失是这种病理的一部分，因此，我们将研究吸入NO或其氧化还原同系物亚硝酸乙酯在肺部炎症治疗中的可能性，以进一步了解参与肺部炎症的分子机制，以及NO如何改变这些机制，我们将检测组织内和灌洗液中活性物质的潜在蛋白质靶标，特别是评估硝化蛋白、S亚硝化蛋白和氧化蛋白的产生。这样的实验可能导致关于反应物种的生理信号通路以及这些通路在炎症过程中如何改变的有价值的信息。最后，我们将研究NO介导的SP-D修饰如何影响纯化的SP-D对表型的挽救。这些实验将为SP-D和NO如何参与维持生理肺功能提供有价值的信息。

DESCRIPTION (provided by applicant): The major goal of this research project is to investigate how NO chemistry plays a role both in the initiation and in the propagation of inflammatory disease within the lung. In particular, we will investigate the effects of altered NO chemistry within an animal model of pulmonary inflammation, ablation of the SP-D gene. Mice lacking the SP-D gene develop a non-specific inflammatory disease within the lung in the absence of an infectious agent. This disease is characterized by parenchymal infiltrates and peribronchial neutrophilial lipoproteinosis, and alterations in the NO chemistry of the lung (loss of S-nitrosothiol, SNO, and gain of nitrotyrosine within tissue and a relative greater increase of nitrates within the bronchiolar alveolar lavage, BAL). Therefore, this animal model provides an ideal opportunity to investigate the role that NO plays in pulmonary inflammation. We will use both pharmacological and genetic interventions to manipulate NO production within this model and examine the effects of these manipulations upon the development of disease. Furthermore, we propose that the loss of normal signaling NO is part of this pathology, therefore, we will investigate the possible use of inhaled NO or its redox congener, ethyl nitrite, in treatment of pulmonary inflammation, in order to further our understanding of the molecular mechanisms involved in pulmonary inflammatory disease, and how NO modifies these mechanisms, we will examine the potential protein targets of reactive species both within the tissue and the lavage, in particular, we will assess the production of nitrated proteins, S-nitrosylated proteins, and oxidized proteins. Such experiments may lead to valuable information concerning the physiological signaling pathways of reactive species and how these pathways are altered during inflammation. Finally, we will examine how rescue of the phenotype by purified SP-D may be affected by NO-mediated modifications of SP-D. These experiments wilt provide valuable information as to how SP-D and NO are involved in the maintenance of physiological pulmonary function.