Nishida T, Nakagawa S, Awata T, Ohashi Y, Watanabe K, Manabe R
J Cell Biol. 1983 Nov;97(5 Pt 1):1653-7. doi: 10.1083/jcb.97.5.1653.
We investigated the effect of fibronectin on epithelial migration onto the stroma in cultured rabbit cornea. Rabbit plasma fibronectin was purified by affinity chromatography using gelatin-Sepharose 4B, and its purity was confirmed by SDS polyacrylamide slab gel electrophoresis. Antibody against rabbit plasma fibronectin raised in guinea pigs formed a single precipitin line against rabbit plasma and purified rabbit plasma fibronectin by Ouchterlony double diffusion test. When rabbit cornea was cut into small blocks and cultured in TCM-199 medium alone, corneal epithelial cells began to migrate on the cut edge of the corneal stroma. The addition of purified rabbit plasma fibronectin to the culture medium significantly enhanced epithelial migration. The degree of enhancement depended on the amount of fibronectin added. When guinea pig IgG anti-rabbit plasma fibronectin was added, epithelial migration was significantly inhibited when compared with that in control cultured corneal blocks. The results demonstrate that fibronectin promotes epithelial migration in the cornea and thus plays an important role in corneal wound healing.
我们研究了纤连蛋白对培养的兔角膜上皮向基质迁移的影响。兔血浆纤连蛋白通过使用明胶 - 琼脂糖凝胶4B的亲和层析法进行纯化,其纯度通过SDS聚丙烯酰胺平板凝胶电泳得以确认。在豚鼠体内产生的抗兔血浆纤连蛋白抗体,通过免疫双扩散试验,与兔血浆和纯化的兔血浆纤连蛋白形成单一沉淀线。当将兔角膜切成小块并单独在TCM - 199培养基中培养时,角膜上皮细胞开始在角膜基质的切割边缘迁移。向培养基中添加纯化的兔血浆纤连蛋白可显著增强上皮迁移。增强程度取决于添加的纤连蛋白量。当添加豚鼠抗兔血浆纤连蛋白IgG时,与对照培养的角膜块相比,上皮迁移受到显著抑制。结果表明,纤连蛋白促进角膜上皮迁移,因此在角膜伤口愈合中起重要作用。