Molecular Regulation of Aging, Tokyo Metropolitan Institute of Gerontology, Tokyo 173-0015, Japan.
Biochem Biophys Res Commun. 2013 Jan 11;430(2):579-84. doi: 10.1016/j.bbrc.2012.11.110. Epub 2012 Dec 7.
Ascorbic acid (AA) is essential for collagen biosynthesis as a cofactor for prolyl and lysyl hydroxylase and as a stimulus for collagen gene expression. Many studies have evaluated the relationship between AA and collagen expression in short- and long-term effects on cells after a single administration of AA into the culture medium. However, no such study has monitored in detail the stability of AA in medium or the alterations of intracellular AA levels during a protracted interval. Therefore, we examined here intracellular AA levels and stability throughout its exposure to human skin fibroblasts in vitro. Moreover, we determined the effects on type 1 and type 4 collagen and sodium-dependent vitamin C transporter (SVCT) gene expression when medium containing 100 μM AA was replaced every 24h for 5 days to avoid depletion of AA. Throughout this long-term culture, intracellular AA levels remained constant; the expression of type 1 and type 4 collagens and SVCT2 mRNA was enhanced, and type 1 procollagen synthesis increased. Thus, these results indicate that human skin fibroblasts exposed to AA over time had rising levels of type 1/type 4 collagens and SVCT2 mRNA expression and type 1 procollagen synthesis.
抗坏血酸(AA)是胶原蛋白生物合成所必需的,作为脯氨酰和赖氨酰羟化酶的辅助因子,以及胶原蛋白基因表达的刺激物。许多研究评估了 AA 对细胞的短期和长期作用后,单次给予培养基中 AA 与胶原蛋白表达之间的关系。然而,尚无研究详细监测培养基中 AA 的稳定性以及细胞内 AA 水平在延长时间间隔内的变化。因此,我们在这里检查了细胞内 AA 水平及其在体外人皮肤成纤维细胞中的暴露稳定性。此外,当含有 100 μM AA 的培养基每 24 小时更换一次以避免 AA 耗尽时,我们确定了对 1 型和 4 型胶原蛋白和钠依赖性维生素 C 转运体(SVCT)基因表达的影响,持续 5 天。在整个长期培养过程中,细胞内 AA 水平保持不变;1 型和 4 型胶原蛋白和 SVCT2 mRNA 的表达增强,1 型前胶原蛋白合成增加。因此,这些结果表明,随着时间的推移,暴露于 AA 的人皮肤成纤维细胞的 1 型/4 型胶原蛋白和 SVCT2 mRNA 表达以及 1 型前胶原蛋白合成水平升高。