肺微环境粘附分子谱对气道上皮／间质修复平衡点调控

申请者提出上皮修复和间质修复的失衡可能是肺纤维化和气道重构的内在原因，粘附分子表达谱及其调控的上皮细胞与间质细胞的相互作用构成平衡调控机制。本项目拟：①利用自主构建的气道上皮整合素β4条件性剔除小鼠，建立损伤模型，观察肺功能、结构、上皮细胞与间质细胞的EMT和MET表型标记，及其与三种主要粘附分子表达谱的对应关系；②在细胞水平，构建3种主要粘附分子不同表达水平的气道上皮细胞系和肺成纤维细胞系，检测各组细胞的增殖、修复指数、EMT和MET表型、胶原合成谱等差异；③利用气道上皮细胞与肺成纤维细胞共培养体系，观察在不同粘附分子表达谱下两种细胞的相互作用，确定相关的分泌型介质；④通过数据分析，寻找有效实现上皮修复与间质修复平衡控制的最佳粘附分子表达水平组合，构建最佳组合转染体系，进行功能验证。该研究将创新肺损伤修复、肺纤维化、气道重构的研究思路，寻求以粘附分子作为治疗上述疾病新靶点的可能性。

The proposal hypothesize that imbalance between epithelial repair and mesenchymal repair possibly contribute the innate cause of pulmonary fibrosis or airway remodelling, and the control mechanism of this repairring equation may involve the interaction between epithelial cells and mesenchymal cells which might be regulated by expression spectrum of adhesive molecules in the pulmonary microenvironment. The project plan: ① A stress injury animal model will be induced on a mouse with integrinβ4 conditional knockout in airway epithelium that has been created in our laborytory, and on this model, ventilation function,lung architecture,some phenotype marker of epithelial-mesenchymal transition (EMT) or mesenchymal-epithelial transition (MET),and their correspond relation with three major kind of adhesive molecules will be observed. ② On cellular level, both airway epithelial celline and pulmonary fibroblast celline with different expression level of three major adhesive molecules will be constructed, and proliferation,repairring index, EMT or MET marker, and difference in collagen synthesis spectrum also be evaluated.③ In a coculture system composed by airway epithelial cells and pulmonary fibroblasts, interaction between two cellline with different adhesive molecules expression spectrum is planed to be investigated and those relevant secretory medium will be idientified.④ From data analysis,an optimal compose of adhesive molecules which can most effetively control the balance between epithelial and mesenchymal repair will be found, and some transinfect system then be designed according to this optimal compose to do functional confirmation. This study will bring a new sight to lung injury repair,pulmonary fibrosis and airway remodelling, and try to seek for new target from adhesive molecules in pulmonary microenvironment for therapy of above lung diseases.