Heino J, Kähäri V M, Jaakkola S, Peltonen J
Department of Medical Biochemistry, University of Turku, Finland.
Matrix. 1989 Jan;9(1):34-9. doi: 10.1016/s0934-8832(89)80016-2.
The synthesis of collagen and its deposition into the extracellular matrix were studied in scleroderma and normal skin fibroblast cultures using immunocytochemistry, electron microscopy and electrophoretic analysis. Indirect immunofluorescence localized type I collagen to intracellular granules of normal skin fibroblasts but not to the extracellular matrix. A 48-h ascorbic acid-treatment of scleroderma and normal skin fibroblasts induced the appearance of fibrillar, collagen-containing deposits in the extracellular space as indicated by indirect immunofluorescence and electron microscopy. Interestingly, similar type I collagen-containing fibrils were observed in cultures of scleroderma skin fibroblasts even without preceding ascorbic acid treatment. Electrophoretic analysis revealed that ascorbic acid increases the conversion of procollagen to collagen to same extent both in scleroderma and control cell layers. The accumulation of collagen to the cell layer of scleroderma fibroblasts was increased when compared to normal skin fibroblasts; whereas the relative conversion of procollagen to collagen was identical in scleroderma and normal fibroblasts. The results show that the conversion of procollagen to collagen in the cell layers of both scleroderma and normal skin fibroblasts is dependent on ascorbic acid, due to improved secretion of procollagen molecules in the presence of ascorbic acid.
利用免疫细胞化学、电子显微镜和电泳分析技术，对硬皮病和正常皮肤成纤维细胞培养物中胶原蛋白的合成及其在细胞外基质中的沉积进行了研究。间接免疫荧光法将I型胶原蛋白定位在正常皮肤成纤维细胞的细胞内颗粒中，而非细胞外基质中。用抗坏血酸处理硬皮病和正常皮肤成纤维细胞48小时后，间接免疫荧光法和电子显微镜观察显示细胞外空间出现了含纤维状胶原蛋白的沉积物。有趣的是，即使在没有预先进行抗坏血酸处理的硬皮病皮肤成纤维细胞培养物中也观察到了类似的含I型胶原蛋白的纤维。电泳分析表明，抗坏血酸在硬皮病和对照细胞层中均能同等程度地促进前胶原向胶原蛋白的转化。与正常皮肤成纤维细胞相比，硬皮病成纤维细胞细胞层中胶原蛋白的积累增加；而硬皮病和正常成纤维细胞中前胶原向胶原蛋白的相对转化率是相同的。结果表明，硬皮病和正常皮肤成纤维细胞细胞层中前胶原向胶原蛋白的转化均依赖于抗坏血酸，这是因为在抗坏血酸存在的情况下前胶原分子的分泌得到了改善。
