Buisson A C, Gilles C, Polette M, Zahm J M, Birembaut P, Tournier J M
INSERM Unité 314, Université de Reims, France.
Lab Invest. 1996 Mar;74(3):658-69.
Wound repair involves many processes including cell migration, provisional matrix deposition, and remodeling. All of these processes are likely to be affected by matrix-modifying enzymes. Members of the matrix metalloproteinases family are physiologic mediators of the extracellular matrix degradation. Within this matrix metalloproteinases family, stromelysins can degrade many components of the extracellular matrix. We therefore tested the hypothesis that stromelysins could be produced by human surface respiratory epithelial (HSRE) cells repairing a wound. Experimental wounds were created in vitro in HSRE cell cultures and in situ in human bronchial mucosa maintained in organ culture. Stromelysin production was measured by casein-gel zymography in cellular protein extracts derived from repairing migratory and nonrepairing stationary cells of wounded HSRE cell cultures. Stromelysin-producing cells present in cell and tissue cultures were localized and characterized using immunofluorescence techniques. Zymographic and immunofluorescence techniques showed that stromelysins were produced exclusively by the migratory HSRE cells. Zymogram analysis showed that stromelysins were overexpressed and overactivated during the wound repair process, with the maximal production observed at wound closure. Using an anti-cytokeratin 14 antibody, we identified stromelysin-3-producing cells as basal epithelial cells. Moreover, most stromelysin-3-producing cells expressed the mesenchymal marker vimentin. Similar to stromelysins localization, vimentin-positive HSRE cells were exclusively located in the wounded area, and they were also positive to cytokeratin 14. In conclusion, stromelysins are suggested to be involved in HSRE cell migration and extracellular matrix remodeling during wound repair. Furthermore, stromelysin production by repairing HSRE cells is linked to the acquisition of a mesenchymal phenotype. HSRE cell migration may then be associated with the shift from an epithelial to a mesenchymal phenotype.
伤口修复涉及许多过程，包括细胞迁移、临时基质沉积和重塑。所有这些过程都可能受到基质修饰酶的影响。基质金属蛋白酶家族的成员是细胞外基质降解的生理介质。在这个基质金属蛋白酶家族中，基质溶解素可以降解细胞外基质的许多成分。因此，我们测试了这样一个假设，即基质溶解素可能由修复伤口的人表面呼吸道上皮（HSRE）细胞产生。在HSRE细胞培养物中体外创建实验性伤口，并在器官培养中维持的人支气管黏膜中原位创建实验性伤口。通过酪蛋白凝胶酶谱法在源自受伤HSRE细胞培养物中修复性迁移细胞和非修复性静止细胞的细胞蛋白提取物中测量基质溶解素的产生。使用免疫荧光技术对细胞和组织培养物中存在的产生基质溶解素的细胞进行定位和表征。酶谱和免疫荧光技术表明，基质溶解素仅由迁移的HSRE细胞产生。酶谱分析表明，基质溶解素在伤口修复过程中过度表达和过度激活，在伤口闭合时观察到最大产量。使用抗细胞角蛋白14抗体，我们将产生基质溶解素-3的细胞鉴定为基底上皮细胞。此外，大多数产生基质溶解素-3的细胞表达间充质标志物波形蛋白。与基质溶解素的定位相似，波形蛋白阳性的HSRE细胞仅位于受伤区域，并且它们对细胞角蛋白14也呈阳性。总之，提示基质溶解素参与伤口修复过程中的HSRE细胞迁移和细胞外基质重塑。此外，修复性HSRE细胞产生基质溶解素与获得间充质表型有关。然后，HSRE细胞迁移可能与从上皮表型向间充质表型的转变有关。
