Geesin J C, Gordon J S, Berg R A
Department of Biochemistry, University of Medicine and Dentistry of New Jersey, Robert Wood Johnson Medical School, Piscataway 08854.
Skin Pharmacol. 1993;6(1):65-71. doi: 10.1159/000211089.
Ascorbic acid has been shown to stimulate collagen synthesis in dermal fibroblasts by increasing the rate of transcription of collagen genes. Experiments involving the use of ascorbic acid require daily supplementation due to the instability of the molecule in aqueous solutions. In order to provide a more stable alternative to ascorbic acid, two salts of ascorbyl-2-phosphate, having a greater chemical stability than ascorbic acid, were tested for their ability to stimulate collagen synthesis in monolayer fibroblast cultures. The concentration and time dependence of their activities were compared with ascorbic acid. The magnesium salt of ascorbyl-2-phosphate was found to be equivalent to ascorbic acid in stimulating collagen synthesis in these assays, while the sodium salt required at least a tenfold greater concentration to produce the same effect as ascorbic acid. Solutions of either ascorbic acid or the ascorbyl-2-phosphate analogs (at 10 mM) in phosphate-buffered saline (PBS) were relatively stable as shown by their decay rates and their ability to stimulate collagen synthesis even after nine days in solution prior to testing their effects on cultured cells. Ascorbic acid was unstable at neutral pH compared to solutions of either sodium or magnesium ascorbyl-2-phosphate. These data support the use of magnesium ascorbyl-2-phosphate in experiments where stability of ascorbic acid is a concern, e.g. in long-term cultures or in in vivo studies.
已证明抗坏血酸可通过提高胶原蛋白基因的转录速率来刺激真皮成纤维细胞中的胶原蛋白合成。由于该分子在水溶液中不稳定，涉及使用抗坏血酸的实验需要每日补充。为了提供一种比抗坏血酸更稳定的替代品，测试了两种抗坏血酸-2-磷酸酯盐（其化学稳定性高于抗坏血酸）在单层成纤维细胞培养物中刺激胶原蛋白合成的能力。将它们活性的浓度和时间依赖性与抗坏血酸进行了比较。在这些试验中发现，抗坏血酸-2-磷酸酯镁盐在刺激胶原蛋白合成方面与抗坏血酸相当，而钠盐产生与抗坏血酸相同效果所需的浓度至少高十倍。抗坏血酸或抗坏血酸-2-磷酸酯类似物（10 mM）在磷酸盐缓冲盐水（PBS）中的溶液相对稳定，这从它们的降解速率以及在测试对培养细胞的影响之前在溶液中放置九天后仍能刺激胶原蛋白合成的能力可以看出。与抗坏血酸-2-磷酸酯钠或镁的溶液相比，抗坏血酸在中性pH下不稳定。这些数据支持在抗坏血酸稳定性是一个问题的实验中使用抗坏血酸-2-磷酸酯镁，例如在长期培养或体内研究中。
