Wilson S E, Schultz G S, Chegini N, Weng J, He Y G
Department of Ophthalmology, University of Texas Southwestern Medical Center at Dallas 75235.
Exp Eye Res. 1994 Jul;59(1):63-71. doi: 10.1006/exer.1994.1081.
The purpose of this study was to determine whether epidermal growth factor (EGF), EGF receptor, transforming growth factor alpha (TGF-alpha), transforming growth factor beta (TGF-beta), acidic fibroblast growth factor (acidic-FGF), basic fibroblast growth factor (basic-FGF), and interleukin-1-alpha (IL-1-alpha) proteins were present in cultures of human corneal cells and/or in sections of human corneal tissue. Immunohistochemistry was performed on human corneal sections. Immunofluorescent cell staining was used to evaluate corneal epithelial, stromal fibroblast, and endothelial cells in primary culture. Basic-FGF production was evaluated in culture cells using immunoprecipitation. EGF, TGF-alpha, TGF-beta-1, and IL-1-alpha were detected by immunohistochemistry in cells in all three layers of the cornea. EGF receptor and acidic FGF were detected by immunohistochemistry in epithelial and endothelial cells, but not in stromal fibroblast cells. Differences in distribution of the growth factors were noted within individual layers of the cornea. EGF and basic-FGF proteins were detected in all three predominant cell types of the cornea using immunocytology. IL-1-alpha protein was detected by immunocytology in corneal epithelial and endothelial cells, but not stromal fibroblasts. Immunoprecipitation confirmed the production of basic-FGF in all three cell types. IL-1-alpha protein detection in the corneal stroma by immunohistology, but not by immunocytology in first passage stromal fibroblasts, suggests that IL-1-alpha may localize to the corneal stroma after production by corneal epithelial and/or endothelial cells.
本研究的目的是确定人角膜细胞培养物和/或人角膜组织切片中是否存在表皮生长因子（EGF）、EGF受体、转化生长因子α（TGF-α）、转化生长因子β（TGF-β）、酸性成纤维细胞生长因子（酸性-FGF）、碱性成纤维细胞生长因子（碱性-FGF）和白细胞介素-1α（IL-1-α）蛋白。对人角膜切片进行免疫组织化学检测。采用免疫荧光细胞染色法评估原代培养的角膜上皮细胞、基质成纤维细胞和内皮细胞。使用免疫沉淀法评估培养细胞中碱性-FGF的产生。通过免疫组织化学在角膜所有三层细胞中检测到EGF、TGF-α、TGF-β-1和IL-1-α。通过免疫组织化学在上皮细胞和内皮细胞中检测到EGF受体和酸性FGF，但在基质成纤维细胞中未检测到。注意到角膜各层内生长因子分布存在差异。使用免疫细胞学法在角膜所有三种主要细胞类型中检测到EGF和碱性-FGF蛋白。通过免疫细胞学法在角膜上皮细胞和内皮细胞中检测到IL-1-α蛋白，但在基质成纤维细胞中未检测到。免疫沉淀法证实所有三种细胞类型均产生碱性-FGF。通过免疫组织学在角膜基质中检测到IL-1-α蛋白，但在第一代基质成纤维细胞中通过免疫细胞学法未检测到，这表明IL-1-α可能在角膜上皮细胞和/或内皮细胞产生后定位于角膜基质。
