Department of Internal Medicine, Respiratory Research Unit, Oulu University Hospital and Institute of Clinical Medicine, University of Oulu, Oulu, Finland.
Lab Invest. 2012 Sep;92(9):1270-84. doi: 10.1038/labinvest.2012.95. Epub 2012 Jun 18.
The characteristic features of myofibroblasts in various lung disorders are poorly understood. We have evaluated the ultrastructure and invasive capacities of myofibroblasts cultured from small volumes of diagnostic bronchoalveolar lavage (BAL) fluid samples from patients with different types of lung diseases. Cells were cultured from samples of BAL fluid collected from 51 patients that had undergone bronchoscopy and BAL for diagnostic purposes. The cells were visualized by transmission electron microscopy and immunoelectron microscopy to achieve ultrastructural localization of alpha-smooth muscle actin (α-SMA) and fibronectin. The levels of α-SMA protein and mRNA and fibronectin mRNA were measured by western blot and quantitative real-time reverse transcriptase polymerase chain reaction. The invasive capacities of the cells were evaluated. The cultured cells were either fibroblasts or myofibroblasts. The structure of the fibronexus, and the amounts of intracellular actin, extracellular fibronectin and cell junctions of myofibroblasts varied in different diseases. In electron and immunoelectron microscopy, cells cultured from interstitial lung diseases (ILDs) expressed more actin filaments and α-SMA than normal lung. The invasive capacity of the cells obtained from patients with idiopathic pulmonary fibrosis was higher than that from patients with other type of ILDs. Cells expressing more actin filaments had a higher invasion capacity. It is concluded that electron and immunoelectron microscopic studies of myofibroblasts can reveal differential features in various diseases. An analysis of myofibroblasts cultured from diagnostic BAL fluid samples may represent a new kind of tool for diagnostics and research into lung diseases.
各种肺部疾病中肌成纤维细胞的特征尚未完全了解。我们评估了从小容积诊断性支气管肺泡灌洗(BAL)液样本中培养的肌成纤维细胞的超微结构和侵袭能力,这些样本来自患有不同类型肺部疾病的患者。通过透射电子显微镜和免疫电镜对细胞进行观察,以实现α-平滑肌肌动蛋白(α-SMA)和纤维连接蛋白的超微结构定位。通过 Western blot 和实时定量逆转录聚合酶链反应测量α-SMA 蛋白和 mRNA 以及纤维连接蛋白 mRNA 的水平。评估细胞的侵袭能力。培养的细胞为成纤维细胞或肌成纤维细胞。纤维连接体的结构以及肌成纤维细胞的细胞内肌动蛋白、细胞外纤维连接蛋白和细胞连接的数量在不同疾病中存在差异。在电子显微镜和免疫电镜下,间质性肺疾病(ILDs)培养的细胞表达的肌动蛋白丝和α-SMA 多于正常肺。特发性肺纤维化患者的细胞侵袭能力高于其他类型 ILD 患者的细胞。表达更多肌动蛋白丝的细胞具有更高的侵袭能力。因此,肌成纤维细胞的电子显微镜和免疫电镜研究可以揭示各种疾病中的差异特征。对诊断性 BAL 液样本中培养的肌成纤维细胞进行分析可能代表了一种用于肺部疾病诊断和研究的新工具。