Wang X, Kamiyama K, Iguchi I, Kita M, Imanishi J
Department of Microbiology, Kyoto Prefectural University of Medicine, Japan.
Invest Ophthalmol Vis Sci. 1994 Nov;35(12):4001-7.
To examine the enhancing effect of cytokines on the corneal epithelial cell migration induced by fibronectin (FN).
A modified Boyden chamber method was used to detect chemotactic cell migration. Cells plated in the inner chamber were incubated with FN, cytokines, or both in the outer chamber at 37 degrees C for 24 hours. Cells that had migrated were stained and counted under a microscope. Checkerboard analysis was used to distinguish chemotaxis from chemokinesis.
FN induced epithelial cell migration, but interleukin (IL)-1 alpha, IL-6, tumor necrosis factor (TNF)-alpha, and epidermal growth factor (EGF) alone did not. These cytokines, even at very low concentrations (0.1 to 100 pg/ml), enhanced FN-induced migration to levels about twofold those observed with FN alone. Checkerboard analysis demonstrated that EGF, but not IL-1 alpha, IL-6, or TNF-alpha, stimulated the chemokinesis of corneal epithelial cells in the presence of FN.
EGF enhanced corneal epithelial cell migration by increasing chemokinesis, whereas IL-1 alpha, IL-6, and TNF-alpha enhanced this migration by increasing the FN-induced chemotactic activity, although these cytokines themselves do not have chemokinetic and chemotactic activity.
研究细胞因子对纤连蛋白（FN）诱导的角膜上皮细胞迁移的增强作用。
采用改良的Boyden小室法检测趋化性细胞迁移。将接种在内室的细胞与外室中的FN、细胞因子或两者一起在37℃孵育24小时。对迁移的细胞进行染色并在显微镜下计数。采用棋盘分析来区分趋化作用和化学增活作用。
FN诱导上皮细胞迁移，但单独的白细胞介素（IL）-1α、IL-6、肿瘤坏死因子（TNF）-α和表皮生长因子（EGF）则不能。这些细胞因子即使在非常低的浓度（0.1至100 pg/ml）下，也能将FN诱导的迁移增强至单独使用FN时观察到的水平的约两倍。棋盘分析表明，在存在FN的情况下，EGF而非IL-1α、IL-6或TNF-α刺激角膜上皮细胞的化学增活作用。
EGF通过增加化学增活作用来增强角膜上皮细胞迁移，而IL-1α、IL-6和TNF-α则通过增加FN诱导的趋化活性来增强这种迁移，尽管这些细胞因子本身不具有化学增活和趋化活性。
