Joyce N C, Meklir B, Neufeld A H
Ophthalmic Pharmacology Unit, Eye Research Institute, Boston, MA 02114.
Invest Ophthalmol Vis Sci. 1990 Sep;31(9):1816-26.
Repair of corneal endothelial wounds involves two forms of cell translocation: (1) "migration," in which individual cells at the wound edge break contacts with neighboring cells and move as individuals into the wound defect, and (2) "spreading," in which cells within the confluent monolayer adjacent to the wound move as a group into the wound area. The authors combined morphometric analysis of Giemsa-stained cultures, phase-contrast video microscopy, and Rh-phalloidin staining of actin filaments to study the effects of epidermal growth factor (EGF) and indomethacin on the migratory and spreading responses to wounding using an in vitro wound-closure model which mimics the amitotic state and general behavior of human corneal endothelium. They found that EGF stimulated the migration of individual cells from the wound edge, induced cellular elongation, and promoted a diffuse distribution of actin filaments. Indomethacin promoted spreading of the confluent monolayer into the wound defect, induced enlargement and flattening of cells, and promoted the formation of long, thick actin stress fibers. These results provide evidence that the migration and spreading responses of corneal endothelial cells to wounding can be pharmacologically separated. The findings suggest that migration of individual cells during wound repair may result from an endogenous form of EGF-like stimulation and that the elongated shape associated with this form of translocation results, at least in part, from an EGF-like alteration in actin-filament organization. Spreading of the confluent monolayer to cover the wound defect may result from a decrease in cyclic adenosine monophosphate induced by a transient reduction in prostaglandin E2 synthesis. This form of translocation may result, in part, from enlargement and flattening of corneal endothelial cells secondary to an enhancement of actin stress-fiber formation.
（1）“迁移”，即伤口边缘的单个细胞与相邻细胞脱离接触，以个体形式移入伤口缺损处；（2）“铺展”，即与伤口相邻的融合单层内的细胞作为一个群体移入伤口区域。作者结合吉姆萨染色培养物的形态计量分析、相差视频显微镜以及肌动蛋白丝的罗丹明 - 鬼笔环肽染色，使用一种模拟人角膜内皮无丝分裂状态和一般行为的体外伤口闭合模型，研究表皮生长因子（EGF）和吲哚美辛对伤口迁移和铺展反应的影响。他们发现，EGF刺激单个细胞从伤口边缘迁移，诱导细胞伸长，并促进肌动蛋白丝的弥漫分布。吲哚美辛促进融合单层向伤口缺损处铺展，诱导细胞增大和平坦化，并促进长而粗的肌动蛋白应力纤维的形成。这些结果证明角膜内皮细胞对伤口的迁移和铺展反应可以通过药理学方法分离。研究结果表明，伤口修复过程中单个细胞的迁移可能源于内源性的类似EGF的刺激，并且这种迁移形式相关的伸长形状至少部分是由肌动蛋白丝组织中类似EGF的改变导致的。融合单层铺展以覆盖伤口缺损可能是由于前列腺素E2合成短暂减少引起的环磷酸腺苷减少所致。这种迁移形式可能部分是由于肌动蛋白应力纤维形成增强导致角膜内皮细胞增大和平坦化所致。
