Poulsen J H, Jensen I M, Petersen U
Department of Clinical Chemistry, University of Aarhus, Denmark.
J Clin Chem Clin Biochem. 1988 Mar;26(3):123-33. doi: 10.1515/cclm.1988.26.3.123.
An in vitro model is presented for the study of glycosaminoglycans in human skin. The synthesis of six glycosaminoglycan species in both dermis and epidermis was measured by D-[3H]glucosamine labelling. Punched biopsies (epidermis + entire dermis) of 3 mm in diameter were cultured at 37 degrees C in 5% carbon dioxide-95% air. When the label was added 18 h after explantation, the incorporation started immediately, and for all glycosaminoglycans the time-dependent incorporation was linear for 16 h. The experimental variation was minimized by expressing the measurements in epidermis "per explant" and in dermis "per mg of wet explant". A ratio to dermal hydroxyproline did not improve the precision. Most of the variation arose "before" isolation and separation of the glycosaminoglycans. The labelled products were macromolecules and were converted to small molecules by chondroitinase ABC + heparinase. The total incorporation in dermis was 2 1/2 times higher than in epidermis. Hyaluronic acid was the predominant synthesized product in dermis, and hyaluronic acid and heparan sulphate were the predominant products in epidermis. The proportions (%) in dermis/epidermis were as follows: hyaluronic acid, 61/44; heparan sulphate, 18/31; dermatan sulphate, 5/8; chondroitin 4/6-sulphate, 10/7 and heparin-like glycosaminoglycan, 1/2. The same species were also demonstrated as native constituents of uncultured human skin. Hyaluronic acid and dermatan sulphate predominated in dermis, whereas no single species predominated in epidermis. Their concentrations in uronic acid equivalents per mg of wet skin (pmol/mg of epidermis + dermis) were as follows in dermis/epidermis: hyaluronic acid, 243/0.48; heparan sulphate, 22/0.44; dermatan sulphate, 170/0.56; chondroitin 4/6-sulphate, 72/0.50; and heparin-like glycosaminoglycan, 5/0.22. Thus, only 0.4% of the in vivo synthesized glycosaminoglycan was present in epidermis.
本文介绍了一种用于研究人类皮肤中糖胺聚糖的体外模型。通过D-[3H]葡萄糖胺标记法测定了真皮和表皮中六种糖胺聚糖的合成情况。将直径为3毫米的打孔活检组织（表皮 + 全层真皮）在37℃、5%二氧化碳 - 95%空气环境中培养。在外植18小时后添加标记物，掺入立即开始，并且对于所有糖胺聚糖，随时间的掺入在16小时内呈线性。通过以“每个外植体”表示表皮中的测量值和以“每毫克湿外植体”表示真皮中的测量值，将实验变异最小化。与真皮羟脯氨酸的比率并未提高精度。大多数变异出现在糖胺聚糖的分离和纯化“之前”。标记产物为大分子，经软骨素酶ABC + 肝素酶作用后转化为小分子。真皮中的总掺入量比表皮高2.5倍。透明质酸是真皮中主要的合成产物，而透明质酸和硫酸乙酰肝素是表皮中的主要产物。真皮/表皮中的比例（%）如下：透明质酸，61/44；硫酸乙酰肝素，18/31；硫酸皮肤素，5/8；硫酸软骨素4/6，10/7；类肝素糖胺聚糖，1/2。相同的糖胺聚糖种类也被证明是未培养的人类皮肤的天然成分。透明质酸和硫酸皮肤素在真皮中占主导，而表皮中没有单一的糖胺聚糖种类占主导。它们在每毫克湿皮肤中以糖醛酸当量表示的浓度（皮摩尔/毫克表皮 + 真皮）在真皮/表皮中如下：透明质酸，243/0.48；硫酸乙酰肝素，22/0.44；硫酸皮肤素，170/0.56；硫酸软骨素4/6，72/0.50；类肝素糖胺聚糖，5/0.22。因此，表皮中仅存在体内合成的糖胺聚糖的0.4%。
