Wang D I, Gotlieb A I
Vascular Research Laboratory, Department of Laboratory Medicine and Pathobiology, Banting and Best Diabetes Centre, The Toronto Hospital Research Institute and University of Toronto, Toronto, Ontario, M5G 2C4, Canada.
Exp Mol Pathol. 1999 Aug;66(3):179-90. doi: 10.1006/exmp.1999.2265.
As endothelial cells convert from quiescent to migrating cells over 8 h along a wound edge, actin microfilaments undergo well-defined sequential changes characterized by an initial random distribution followed by a parallel and then a perpendicular orientation of microfilaments with respect to the wound edge. The latter is associated with subsequent cell migration. We tested the hypothesis that fibroblast growth factor 2 (FGF-2) can enhance the very early stages of wound repair even prior to migration and that FGF-2 enhancement of wound repair is associated with changes in the endothelial actin cytoskeleton. Using an in vitro two-sided wound model, the addition of FGF-2 at the time of wounding enhanced the extent of wound closure over 8 h. Treatment with FGF-2 was associated with significantly longer cells along the wound edge at 4 and 8 h after wounding. When treated with increasing concentrations of neutralizing FGF-2 antibody, the extent of wound closure decreased over 8 h and was associated with a decrease in cell length along the wound edge. Actin microfilaments were localized using rhodamine phalloidin and viewed using laser confocal microscopy. At 4 h after wounding, FGF-2 treatment was associated with significantly more cells along the wound expressing perpendicular microfilaments compared to untreated cells, which suggested a more rapid transition of parallel to perpendicular microfilament distribution. Thus, FGF-2 affects the very early stages of wound repair prior to migration by enhancing wound closure due to the early appearance of perpendicular microfilaments and lengthening of cells along the wound edge.
在内皮细胞沿着伤口边缘在8小时内从静止细胞转变为迁移细胞的过程中，肌动蛋白微丝会经历明确的顺序变化，其特征是最初随机分布，随后微丝相对于伤口边缘呈平行排列，然后呈垂直排列。后者与随后的细胞迁移相关。我们测试了以下假设：成纤维细胞生长因子2（FGF-2）甚至在迁移之前就能增强伤口修复的非常早期阶段，并且FGF-2对伤口修复的增强作用与内皮肌动蛋白细胞骨架的变化有关。使用体外双侧伤口模型，在受伤时添加FGF-2可在8小时内增强伤口闭合的程度。FGF-2处理与受伤后4小时和8小时伤口边缘细胞明显更长有关。当用浓度不断增加的中和FGF-2抗体处理时，伤口闭合程度在8小时内降低，并且与伤口边缘细胞长度的减少有关。使用罗丹明鬼笔环肽定位肌动蛋白微丝，并使用激光共聚焦显微镜观察。与未处理的细胞相比，受伤后4小时，FGF-2处理使伤口边缘表达垂直微丝的细胞明显更多，这表明从平行微丝分布到垂直微丝分布的转变更快。因此，FGF-2通过增强伤口闭合来影响迁移之前伤口修复的非常早期阶段，这是由于垂直微丝的早期出现以及伤口边缘细胞的延长。
