嗜酸粒细胞性支气管炎小鼠模型的建立和支气管哮喘气道高反应性发生机制的研究

Airway hypertension (AHR) is the key characteristic of asthma, but the pathogenesis of AHR is unclear till now because of complex inter relation. Eosinophilic bronchitis (EB) patients have same symptoms of chronic cough, eosinophilic airway inflammation and effective treatment of steroid as asthma patients, but have not AHR which is different from ashtma ones. EB supplies a valuable control tool for asthma AHR mechanism study as it has similar backgroud of other features.Suitable animal model is very important for pathogenesis research, but now there is no concrete EB animal model established through the world. Our team has established and reported the detection method of mouse cough and the mouse model with airway eosinophilic inflammation but no AHR. And our former studies found some differential proteins in EB and asthma patients and mouse models by proteomics analysis.Now we will establish a developed EB mouse model with its four characterics mentioned before. Then BALF of EB and asthma patients and lung tissues of EB and asthma mouses will be gathered for studies of differential proteins by developed iTRAQ proteomics technique. Synthesized analysis of animal and human beings results should be carried out for research of ashtma AHR related biological markers, which will supply scientific basis for mechanism studies of AHR and diagnosis and therapy of ashtma.

支气管哮喘的关键特征是气道高反应性（AHR），但AHR发生机制复杂，至今尚不清楚。嗜酸粒细胞性支气管炎（EB）同哮喘一样具有慢性咳嗽、气道嗜酸粒细胞性炎症、激素治疗有效特点，但无AHR存在。因此在背景一致基础上EB为哮喘AHR发生机制研究提供了良好参照。合适的动物模型对阐明发病机理十分必要，但目前国际上尚未建立EB动物模型。本课题组前期工作在国际上首先建立了有气道炎症但无AHR的类EB小鼠模型和小鼠咳嗽检测方法。本项目将在此基础上进一步建立具有全部四个临床特征的全EB小鼠模型；继而使用EB和哮喘小鼠肺组织标本，应用目前最先进的iTRAQ蛋白质组学技术，判断在AHR发生机制中起关键作用的靶向蛋白谱；针对上调蛋白使用蛋白阻断剂，下调蛋白进行基因转入，或同时结合两种方法影响蛋白表达，观察小鼠气道反应性变化，最终确定AHR发生机制中的关键蛋白，为临床哮喘患者研究及诊疗提供靶向目标。

支气管哮喘的关键特征是气道高反应性（AHR），但AHR发生机制复杂，至今尚不清楚。嗜酸粒细胞性支气管炎（EB）同哮喘一样具有慢性咳嗽、气道嗜酸粒细胞性炎症、激素治疗有效特点，但无AHR存在。因此在背景一致基础上EB为哮喘AHR发生机制研究提供了良好参照。合适的动物模型对阐明发病机理十分必要，本课题组在国际上首先建立了咳嗽明显， EOS气道炎症，无气道高反应性，糖皮质激素治疗有效的EB小鼠模型。由于支气管哮喘涉及很多与分子遗传、生化途径及其与环境因素的相互作用。因此哮喘表型的复杂本质同遗传异质性及环境的影响使得对其具体机制的研究遇到了很多困难，仅从基因水平对其研究所取得的成果十分有限。蛋白质主要负责生物功能和生理、病理表型的表达，人体内一些非细胞成分，如血浆和肺上皮细胞衬液，虽然只有少量的DNA或RNA，却有着大量的蛋白质，这些蛋白质可以作为疾病的生物标志物。我们利用建立的EB小鼠模型，在总体背景均一、仅有气道反应性差异基础上，与哮喘小鼠进行差异蛋白的比较，发现EB和哮喘＞1.3倍的上调差异蛋白有14个，下调差异蛋白有50个，并对其中最可能是目标蛋白的差异蛋白（如Myh6,S100A8, S100A9等）进行了验证。在一定程度上阐明了EB和哮喘的蛋白质调控网络，为哮喘的早期干预提供有力依据。