Malmström J, Westergren-Thorsson G, Marko-Varga G
BMC Cell & Molecular Biology, University of Lund, Sweden.
Electrophoresis. 2001 May;22(9):1776-84. doi: 10.1002/1522-2683(200105)22:9<1776::AID-ELPS1776>3.0.CO;2-H.
Subepithelial fibrosis in asthma involves an increase in the thickening of the lamina reticularis and is due to increased deposition of collagen I, III and V, and fibronectin. The cause of the thickening of the reticular layer is not known in detail, however, it is proposed to be caused by bronchial myofibroblasts. The transformation of fibroblasts to myofibroblasts may be contributed by inflammatory cytokines. In this paper we have studied and compared in vivo tissue material with a human fibroblast target cell. A normal primary fetal fibroblast cell line and HFL-1 (human fibroblast lurg cells) were used as a comparison between fibroblasts from human central biopsies regarding morphology and cell proliferation. Both cell morphology and cell proliferation rate was similar between the different set of cell cultures. Furthermore, it could be concluded that fibroblasts cultures from patients with asthma were surrounded by more extracellular matrix molecules compared to the primary cell line HFL-1, which may mimic the in vivo situation during formation of fibrosis. We wanted to investigate if differential protein display by two-dimensional (2-D) gel electrophoresis and subsequent protein identification by matrix assisted laser desorption/ionization-time of flight (MALDI-TOF)-mass spectrometry could reveal proteins induced by cytokine stimulation that can be correlated to the transformation of normal human fetal lungs cells into a more myofibroblast like phenotype. After stimulation with transforming growth factor-beta (TGF-beta) several myofibroblast markers were found to be regulated. Especially cytoskeletal and cytoskeletal-associated proteins like actin isoforms and tropomyosin, proteins that are responsible for contraction as well as transportation of extra cellular matrix proteins, which are overproduced in the formation of fibrosis. These results indicate that TGF-beta, which is increased in a fibrotic process, participates in the transformation of fibroblasts to myofibroblasts.
哮喘中的上皮下纤维化涉及网状板增厚增加，这是由于I型、III型和V型胶原蛋白以及纤连蛋白的沉积增加所致。网状层增厚的原因尚不完全清楚，但据推测是由支气管肌成纤维细胞引起的。成纤维细胞向肌成纤维细胞的转化可能由炎性细胞因子促成。在本文中，我们研究并比较了体内组织材料与人类成纤维细胞靶细胞。使用正常的原代胎儿成纤维细胞系和HFL-1（人成纤维细胞鲁格细胞）来比较来自人类中心活检组织的成纤维细胞在形态和细胞增殖方面的情况。不同组细胞培养物之间的细胞形态和细胞增殖率相似。此外，可以得出结论，与原代细胞系HFL-1相比，哮喘患者的成纤维细胞培养物被更多的细胞外基质分子包围，这可能模拟了纤维化形成过程中的体内情况。我们想研究二维（2-D）凝胶电泳的差异蛋白质展示以及随后通过基质辅助激光解吸/电离飞行时间（MALDI-TOF）质谱进行的蛋白质鉴定是否能够揭示由细胞因子刺激诱导的、可与正常人胎儿肺细胞向更肌成纤维细胞样表型转化相关的蛋白质。在用转化生长因子-β（TGF-β）刺激后，发现几种肌成纤维细胞标志物受到调控。特别是细胞骨架和细胞骨架相关蛋白，如肌动蛋白异构体和原肌球蛋白，这些蛋白负责收缩以及细胞外基质蛋白的运输，而细胞外基质蛋白在纤维化形成过程中过度产生。这些结果表明，在纤维化过程中增加的TGF-β参与了成纤维细胞向肌成纤维细胞的转化。
