Shock A, Rabe K F, Dent G, Chambers R C, Gray A J, Chung K F, Barnes P J, Laurent G J
Department of Thoracic Medicine, National Heart and Lung Institute, University of London, UK.
Clin Exp Immunol. 1991 Oct;86(1):185-90. doi: 10.1111/j.1365-2249.1991.tb05793.x.
Eosinophils have been implicated in several disorders associated with the development of fibrosis. This led us to investigate the interactions between eosinophils and fibroblasts in vitro. Adhesion between purified guinea pig peritoneal eosinophils and monolayers of human fetal lung fibroblasts was assessed using the rose bengal dye staining assay. Fibroblast replication was assessed using a colorimetric assay based upon the uptake and subsequent release of methylene blue. Addition of phorbol myristate acetate induced a rapid, time-dependent increase in eosinophil adhesion (127% and 328% over basal adhesion after 10 and 30 min, respectively). Phorbol myristate acetate-induced adhesion was inhibited by the peptides RGDS and GRGDS (48% and 42%, respectively using 1 mM peptide) and by nordihydroguaiaretic acid, an inhibitor of the lipoxygenase pathway of arachidonic acid metabolism (46% inhibition at 15 microM). In addition, 24 h culture of fibroblast monolayers with interleukin 1 alpha (IL-1 alpha) or tumour necrosis factor alpha (TNF alpha) resulted in enhanced adhesion (10 U/ml IL-1 alpha stimulated adhesion by 55% of control, 500 U/ml TNF alpha by 75% of control). Conditioned media from cultured eosinophils stimulated fibroblast replication in a time-dependent fashion with maximal stimulation at 3 h. In contrast, media from guinea pig peritoneal macrophages in culture did not show such an effect. This study indicates that eosinophils are capable of both adhering to and releasing mitogens for fibroblasts in vitro. These observations suggest that eosinophils have the capacity to play a role in the development of fibrosis in disorders where they have been shown to be present.
嗜酸性粒细胞与多种纤维化相关疾病有关。这促使我们在体外研究嗜酸性粒细胞与成纤维细胞之间的相互作用。使用孟加拉玫瑰红染料染色试验评估纯化的豚鼠腹腔嗜酸性粒细胞与人胎儿肺成纤维细胞单层之间的黏附。使用基于亚甲蓝摄取和随后释放的比色法评估成纤维细胞增殖。添加佛波酯肉豆蔻酸酯可诱导嗜酸性粒细胞黏附迅速、随时间增加（分别在10分钟和30分钟后比基础黏附增加127%和328%）。佛波酯肉豆蔻酸酯诱导的黏附受到肽RGDS和GRGDS（分别使用1 mM肽时抑制48%和42%）以及去甲二氢愈创木酸（花生四烯酸代谢脂氧合酶途径的抑制剂，在15 microM时抑制46%）的抑制。此外，用白细胞介素1α（IL-1α）或肿瘤坏死因子α（TNFα）对成纤维细胞单层进行24小时培养可增强黏附（10 U/ml IL-1α刺激黏附比对照增加55%，500 U/ml TNFα刺激黏附比对照增加75%）。来自培养的嗜酸性粒细胞的条件培养基以时间依赖性方式刺激成纤维细胞增殖，在3小时时达到最大刺激。相比之下，豚鼠腹腔巨噬细胞培养的培养基未显示出这种作用。这项研究表明，嗜酸性粒细胞在体外既能黏附于成纤维细胞，又能释放促有丝分裂原。这些观察结果表明，在已证明嗜酸性粒细胞存在的疾病中，它们有能力在纤维化发展中发挥作用。
