Mohay J, McLaughlin B J
Department of Ophthalmology & Visual Sciences, Kentucky Lions Eye Research Institute, University of Lousville School of Medicine 40292, USA.
Graefes Arch Clin Exp Ophthalmol. 1995 Nov;233(11):727-36. doi: 10.1007/BF00164678.
The aim of the present study was to compare the morphology, proliferative activity and cytoskeletal organization of bovine corneal endothelial cells during wound healing under normal and mitotically inhibited conditions.
Cell cultures were grown to confluency and incubated with the mitotic inhibitor 5-fluorouracil (5-FU; 2.5 micrograms/ml) followed by a touch wound. Control cultures were maintained without 5-FU. Mitotic activity, F-actin, vinculin, vimentin and connexin 43 localization were evaluated before, during and after wound closure.
5-FU inhibited irreversibly the mitotic activity of corneal endothelial cells during the whole wound healing process. In the presence of 5-FU, a high degree of polymegathism and delay in actin and vinculin redistribution to the cell borders after wound closure was observed. Vimentin and connexin 43 immunolabeling revealed only slight differences between 5-FU-treated and control cultures.
Significant changes in cell geometry and cytoskeletal organization in the amitotic corneal endothelium became manifested only after wounding. These changes may influence cell-cell and cell-matrix interactions as well as functional restoration of the monolayer after wound closure.
本研究的目的是比较正常和有丝分裂受抑制条件下牛角膜内皮细胞在伤口愈合过程中的形态、增殖活性和细胞骨架组织。
将细胞培养至汇合状态，用有丝分裂抑制剂5-氟尿嘧啶（5-FU；2.5微克/毫升）处理，随后进行划痕损伤。对照培养物不添加5-FU。在伤口闭合前、期间和之后评估有丝分裂活性、F-肌动蛋白、纽蛋白、波形蛋白和连接蛋白43的定位。
在整个伤口愈合过程中，5-FU不可逆地抑制角膜内皮细胞的有丝分裂活性。在存在5-FU的情况下，观察到高度的大小不均一性以及伤口闭合后肌动蛋白和纽蛋白重新分布到细胞边界的延迟。波形蛋白和连接蛋白43免疫标记显示，5-FU处理的培养物与对照培养物之间仅有轻微差异。
无丝分裂的角膜内皮细胞在伤口愈合后，细胞几何形状和细胞骨架组织发生显著变化。这些变化可能影响细胞间和细胞与基质的相互作用，以及伤口闭合后单层细胞的功能恢复。
