Varani J, Mitra R S, Gibbs D, Phan S H, Dixit V M, Mitra R, Wang T, Siebert K J, Nickoloff B J, Voorhees J J
Department of Pathology, University of Michigan Medical School, Ann Arbor 48109.
J Invest Dermatol. 1990 May;94(5):717-23. doi: 10.1111/1523-1747.ep12876294.
All-trans retinoic acid was examined for effects on human dermal fibroblast proliferation and for effects on fibroblast production and expression of non-collagenous and collagenous components of the extracellular matrix in vitro. Fibroblast proliferation was blocked when the cells were cultured in the presence of a serum-free culture medium containing epidermal growth factor, hydrocortisone, insulin, ethanolamine, phosphoethanolamine, and bovine pituitary extract as growth supplements and 0.15 mM Ca++. This level of extracellular Ca++ is lower than that needed to support fibroblast growth. Under these conditions, growth was stimulated by all-trans retinoic acid. Proliferation was also stimulated in the same basal medium without the growth supplements. Growth-promoting concentrations of all-trans retinoic acid ranged from 0.5-2.0 micrograms/ml (1.7-6.6 X 10(-6) M). Stimulation of proliferation was not seen at higher or lower concentrations. Concentrations of all-trans retinoic acid that stimulated proliferation also induced increased production of fibronectin as indicated by biosynthetic labeling/immunoprecipitation and by enzyme-linked immunosorbent assay. Increased production was associated with increased staining for fibronectin in the extracellular matrix. Increased production of two other non-collagenous extracellular matrix component, i.e., thrombospondin and laminin, also occurred in all-trans retinoic acid-treated cells. At 0.5 micrograms/ml, all-trans retinoic acid also stimulated production of type I collagen by the dermal fibroblasts, but at higher concentrations (2.5 micrograms/ml) production of type I collagen was inhibited. These data indicate that all-trans retinoic acid can induce changes in dermal fibroblasts in vitro (i.e., increased proliferation and extracellular matrix production) that mimic the major changes seen in the dermis after topical treatment with this agent.
研究了全反式维甲酸对人皮肤成纤维细胞增殖的影响，以及对体外成纤维细胞产生和表达细胞外基质中非胶原蛋白和胶原蛋白成分的影响。当细胞在含有表皮生长因子、氢化可的松、胰岛素、乙醇胺、磷酸乙醇胺和牛垂体提取物作为生长补充剂以及0.15 mM钙离子的无血清培养基中培养时，成纤维细胞增殖受到抑制。这种细胞外钙离子水平低于支持成纤维细胞生长所需的水平。在这些条件下，全反式维甲酸可刺激生长。在没有生长补充剂的相同基础培养基中，增殖也受到刺激。促进生长的全反式维甲酸浓度范围为0.5 - 2.0微克/毫升（1.7 - 6.6×10⁻⁶ M）。在更高或更低浓度下未观察到增殖刺激。通过生物合成标记/免疫沉淀和酶联免疫吸附测定表明，刺激增殖的全反式维甲酸浓度也诱导纤连蛋白产生增加。产生增加与细胞外基质中纤连蛋白染色增加相关。在全反式维甲酸处理的细胞中，另外两种非胶原蛋白细胞外基质成分，即血小板反应蛋白和层粘连蛋白的产生也增加。在0.5微克/毫升时，全反式维甲酸也刺激真皮成纤维细胞产生I型胶原蛋白，但在更高浓度（2.5微克/毫升）时，I型胶原蛋白的产生受到抑制。这些数据表明，全反式维甲酸可在体外诱导真皮成纤维细胞发生变化（即增殖增加和细胞外基质产生增加），这与局部应用该药物后真皮中所见的主要变化相似。
