The Pulmonary Center, Boston University School of Medicine, Boston, Massachusetts, United States of America.
PLoS One. 2013 Sep 9;8(9):e74469. doi: 10.1371/journal.pone.0074469. eCollection 2013.
Phenotypes of lung smooth muscle cells in health and disease are poorly characterized. This is due, in part, to a lack of methodologies that allow for the independent and direct isolation of bronchial smooth muscle cells (BSMCs) and vascular smooth muscle cells (VSMCs) from the lung. In this paper, we describe the development of a bi-fluorescent mouse that permits purification of these two cell populations by cell sorting. By subjecting this mouse to an acute allergen based-model of airway inflammation that exhibits many features of asthma, we utilized this tool to characterize the phenotype of so-called asthmatic BSMCs. First, we examined the biophysical properties of single BSMCs from allergen sensitized mice and found increases in basal tone and cell size that were sustained ex vivo. We then generated for the first time, a comprehensive characterization of the global gene expression changes in BSMCs isolated from the bi-fluorescent mice with allergic airway inflammation. Using statistical methods and pathway analysis, we identified a number of differentially expressed mRNAs in BSMCs from allergen sensitized mice that code for key candidate proteins underlying changes in matrix formation, contractility, and immune responses. Ultimately, this tool will provide direction and guidance for the logical development of new markers and approaches for studying human lung smooth muscle.
肺平滑肌细胞在健康和疾病中的表型特征尚未得到充分描述。这部分归因于缺乏能够独立且直接从肺部分离支气管平滑肌细胞（BSMCs）和血管平滑肌细胞（VSMCs）的方法。本文中，我们描述了一种双荧光小鼠的开发，该小鼠可通过细胞分选来纯化这两种细胞群。通过使该小鼠经受表现出许多哮喘特征的急性变应原气道炎症模型，我们利用该工具来表征所谓的哮喘 BSMC 的表型。首先，我们研究了来自变应原致敏小鼠的单个 BSMC 的生物物理特性，发现基础张力和细胞大小增加，并在体外持续存在。然后，我们首次全面表征了来自具有变应性气道炎症的双荧光小鼠中分离的 BSMC 的全基因表达变化。使用统计方法和途径分析，我们在变应原致敏小鼠的 BSMC 中鉴定出许多差异表达的 mRNAs，这些 mRNAs 编码构成基质形成、收缩性和免疫反应变化的关键候选蛋白。最终，该工具将为研究人类肺平滑肌的新标志物和方法的逻辑发展提供方向和指导。
