Buisson A C, Zahm J M, Polette M, Pierrot D, Bellon G, Puchelle E, Birembaut P, Tournier J M
Unité INSERM 314, Université de Reims, France.
J Cell Physiol. 1996 Feb;166(2):413-26. doi: 10.1002/(SICI)1097-4652(199602)166:2<413::AID-JCP20>3.0.CO;2-A.
Following epithelial injury, extracellular matrix undergoes imposing remodelings. We examined the contribution of matrix metalloproteinases, gelatinases A and B, in an in vitro wound repair model of human respiratory epithelium. Confluent human surface respiratory epithelial (HSRE) cells cultured from dissociated surface cells of human nasal polyps were chemically injured. Over the next 3 to 5 days, cells migrated onto the injured area to repair the circular wound. Repair kinetics of these wounds was monitored until wound closure occurred. Gelatinolytic activities were analysed in culture supernates and in cell protein extracts derived from repairing migratory and non repairing stationary cells. Small amounts of gelatinase A were expressed by HSRE cells, and variations of this gelatinase remained very weak for the time of the wound repair. In contrast, gelatinase B was upregulated during the wound repair process, with a maximum peak observed at wound closure. A marked gelatinase B activation occurred only in cells involved in the repair process. Gelatinase B was localized in some migratory basal cells, recognized by an anti-cytokeratin 14 antibody and located around the wound. We could not detect any gelatinase A in repairing or in stationary HSRE cells. Addition of the 6-6B monoclonal antibody, known to inhibit gelatinase B activation, to the culture medium during the repair process resulted in a dose-dependent decrease of the wound repair speed. These results suggest that gelatinase B, produced by epithelial cells, actively contributes to the wound repair process of the respiratory epithelium.
上皮损伤后，细胞外基质会发生显著重塑。我们在人呼吸道上皮的体外伤口修复模型中研究了基质金属蛋白酶、明胶酶A和B的作用。从人鼻息肉的分离表面细胞培养的汇合人表面呼吸道上皮（HSRE）细胞受到化学损伤。在接下来的3至5天内，细胞迁移到损伤区域以修复圆形伤口。监测这些伤口的修复动力学，直至伤口闭合。分析培养上清液以及来自修复迁移细胞和非修复静止细胞的细胞蛋白提取物中的明胶酶活性。HSRE细胞表达少量明胶酶A，在伤口修复期间这种明胶酶的变化仍然非常微弱。相比之下，明胶酶B在伤口修复过程中上调，在伤口闭合时观察到最大峰值。明显的明胶酶B激活仅发生在参与修复过程的细胞中。明胶酶B定位于一些迁移的基底细胞中，这些细胞可被抗细胞角蛋白14抗体识别并位于伤口周围。在修复过程中，我们在修复或静止的HSRE细胞中均未检测到任何明胶酶A。在修复过程中向培养基中添加已知可抑制明胶酶B激活的6-6B单克隆抗体，导致伤口修复速度呈剂量依赖性降低。这些结果表明，上皮细胞产生的明胶酶B积极参与呼吸道上皮的伤口修复过程。
