Schmidt A, Lemming G, Yoshida K, Buddecke E
Institut für Arterioskleroseforschung an der Universität Münster, Deutschland.
Eur J Cell Biol. 1992 Dec;59(2):322-8.
Heparan sulfate isolated from mammalian arterial tissue inhibits the growth of homologous arterial smooth muscle cells when added to subconfluent cell cultures at a concentration of 50 to 100 micrograms/ml culture medium. Disintegration of the heparan sulfate molecule by hydrazinolysis that deacetylates N-acetylglucosaminyl residues and by subsequent treatment with nitrous acid at pH 3.9 results in the formation of a mixture of oligosaccharides which was further resolved into sulfate-enriched oligosaccharides with antiproliferative activity in an in vitro bioassay system. A decasaccharide and dodeca/tetradecasaccharide fraction had a significantly higher antiproliferative effect on arterial smooth muscle cells than the native heparan sulfate molecule. The antiproliferative oligosaccharides have a sulfate content of 0.9 to 1.2 sulfate groups/disaccharide unit and consist of 60 to 70% monosulfated, disulfated, and trisulfated disaccharide units. Up to 32% of the sulfate groups were in 2-position of the uronic acid. In contrast, nitrous acid degradation of heparan sulfate at pH 1.5, which cleaves glycosidic linkages of N-sulfoglucosaminyl residues, results in the formation of sulfate-poor or sulfate-free oligosaccharides without antiproliferative potency. The results indicate that (a) heparan sulfate has a heterogeneous molecular organization where sulfate-rich domains are separated by sulfate-poor sequences and that (b) the antiproliferative activity of heparan sulfate resides in domains enriched with 2-O-sulfated uronic acid residues.
从哺乳动物动脉组织中分离出的硫酸乙酰肝素，当以50至100微克/毫升培养基的浓度添加到亚汇合细胞培养物中时，可抑制同源动脉平滑肌细胞的生长。通过肼解作用使N - 乙酰葡糖胺残基脱乙酰化，随后在pH 3.9下用亚硝酸处理，使硫酸乙酰肝素分子分解，形成寡糖混合物，该混合物在体外生物测定系统中进一步分解为具有抗增殖活性的富含硫酸盐的寡糖。一种十糖和十二/十四糖级分对动脉平滑肌细胞的抗增殖作用明显高于天然硫酸乙酰肝素分子。抗增殖寡糖的硫酸盐含量为0.9至1.2个硫酸盐基团/二糖单位，由60至70%的单硫酸化、双硫酸化和三硫酸化二糖单位组成。高达32%的硫酸盐基团位于糖醛酸的2位。相比之下，在pH 1.5下硫酸乙酰肝素的亚硝酸降解会切断N - 磺基葡糖胺残基的糖苷键，导致形成缺乏抗增殖能力的贫硫酸盐或无硫酸盐的寡糖。结果表明：(a)硫酸乙酰肝素具有异质分子结构，其中富含硫酸盐的结构域被贫硫酸盐序列隔开；(b)硫酸乙酰肝素的抗增殖活性存在于富含2 - O - 硫酸化糖醛酸残基的结构域中。
