Wygrecka M, Markart P, Ruppert C, Petri K, Preissner K T, Seeger W, Guenther A
Department of Biochemistry, Faculty of Medicine, University of Giessen Lung Center, Klinikstr. 36, 35392 Giessen, Germany.
Eur Respir J. 2007 Jun;29(6):1105-14. doi: 10.1183/09031936.00097306. Epub 2007 Mar 1.
Excessive pro-coagulant and decreased fibrinolytic activities in the alveolar compartment have been repeatedly documented for inflammatory and fibrotic lung diseases. The current authors determined the contribution of different resident lung cells to the altered local production of coagulation- and fibrinolysis-system components in bleomycin-injured mouse lungs via cell-specific and quantitative assessment of mRNA levels of various pro-coagulant and (anti)-fibrinolytic factors. Laser-assisted microdissection technology was used to sample specific cell populations in combination with subsequent mRNA analysis by real-time quantitative reverse transcriptase-PCR. Additionally, western blot analysis, immunohistochemistry and activity assays were performed. Following bleomycin challenge, the strongest induction of tissue factor and plasminogen activator inhibitor (PAI)-1 mRNA expression was observed in alveolar macrophages (approximately 250- and 60-fold induction, respectively). These factors were also upregulated in alveolar type II cells, but to an approximately six-fold lesser extent. In contrast, PAI-2 expression was induced exclusively in alveolar macrophages. A slight increase of urokinase-type plasminogen activator (uPA) expression was also observed in alveolar macrophages (two-fold induction), but uPA activity was reduced due to a disproportionate increase of PAI production. Alveolar macrophages and, to a lesser extent, alveolar type II cells are the main sources of locally produced pro-coagulant and anti-fibrinolytic factors in bleomycin-injured lungs.
肺泡腔中促凝活性过高和纤溶活性降低在炎症性和纤维化性肺病中已被反复证实。本文作者通过对各种促凝和（抗）纤溶因子mRNA水平进行细胞特异性和定量评估，确定了不同的肺固有细胞对博来霉素损伤的小鼠肺中凝血和纤溶系统成分局部产生改变的作用。激光辅助显微切割技术用于对特定细胞群体进行采样，并结合随后的实时定量逆转录-聚合酶链反应进行mRNA分析。此外，还进行了蛋白质印迹分析、免疫组织化学和活性测定。博来霉素攻击后，在肺泡巨噬细胞中观察到组织因子和纤溶酶原激活物抑制剂（PAI）-1 mRNA表达的最强诱导（分别约为250倍和60倍诱导）。这些因子在肺泡II型细胞中也上调，但程度约小6倍。相比之下，PAI-2表达仅在肺泡巨噬细胞中被诱导。在肺泡巨噬细胞中也观察到尿激酶型纤溶酶原激活物（uPA）表达略有增加（2倍诱导），但由于PAI产生的不成比例增加，uPA活性降低。肺泡巨噬细胞以及程度较轻的肺泡II型细胞是博来霉素损伤肺中局部产生的促凝和抗纤溶因子的主要来源。
