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硫酸软骨素和硫酸乙酰肝素对草酸钙结晶的单独作用:结石形成者与正常对照受试者之间的差异

Separate effects of urinary chondroitin sulphate and heparan sulphate on the crystallization of urinary calcium oxalate: differences between stone formers and normal control subjects.

作者信息

Shum D K, Gohel M D

机构信息

Department of Biochemistry, Faculty of Medicine, University of Hong Kong.

出版信息

Clin Sci (Lond). 1993 Jul;85(1):33-9. doi: 10.1042/cs0850033.

DOI:10.1042/cs0850033
PMID:8149691
Abstract
  1. Urinary glycosaminoglycans were recovered from the papain digest of polyanions precipitated sequentially by cetylpyridinium chloride and sodium acetate-saturated ethanol. Those from the early morning urine of 48 stone formers and 43 normal control subjects measured 11 and 16 micrograms of uronic acid/ml of urine, respectively. 2. Preparative agarose gel electrophoresis of the recovered glycosaminoglycans in barium acetate buffer (pH 5.8) yielded fractions containing purely chondroitin sulphate, co-polymeric chondroitin/dermatan sulphates and heparan sulphate. Identification was based on the susceptibility of the fractions to chondroitinase or nitrous acid treatment. Similar compositions of glycosaminoglycan classes were observed in samples from stone formers and normal control subjects. 3. The fractionated glycosaminoglycans were dissolved in urine ultrafiltrate to assay for nucleation-promoting and growth-inhibiting activities towards crystallization of urinary calcium oxalate. When compared at the same uronic acid concentration, both the urinary chondroitin sulphate isomers and heparan sulphates of stone formers demonstrated the capacity to enhance crystal nucleation from calcium oxalate endogenous in urine ultrafiltrates, whereas only urinary heparan sulphates of normal control subjects demonstrated this capacity. 4. Tissue-derived reference chondroitin sulphate, dermatan sulphate and heparin, when similarly tested, showed negligible crystal nucleation-promoting activity. The tissue-derived heparan sulphate was similar to the urinary heparan sulphates in showing marked crystal nucleation-promoting activity. 5. Crystal-growth inhibitory activity was evident in all urinary glycosaminoglycan fractions studied. In particular, urinary heparan sulphate of normal control subjects showed higher activity than that of stone formers or the chondroitin sulphate isomers of both stone formers and normal control subjects (P < 0.005).
摘要
  1. 从依次用十六烷基氯化吡啶和醋酸钠饱和乙醇沉淀的聚阴离子的木瓜蛋白酶消化物中回收尿糖胺聚糖。48名结石形成者和43名正常对照受试者晨尿中的尿糖胺聚糖分别为每毫升尿液11微克和16微克糖醛酸。2. 在醋酸钡缓冲液(pH 5.8)中对回收的糖胺聚糖进行制备性琼脂糖凝胶电泳,得到纯硫酸软骨素、硫酸软骨素/硫酸皮肤素共聚物和硫酸乙酰肝素的组分。鉴定基于这些组分对软骨素酶或亚硝酸处理的敏感性。在结石形成者和正常对照受试者的样本中观察到类似的糖胺聚糖类别组成。3. 将分级分离的糖胺聚糖溶解在尿液超滤液中,以测定其对草酸钙结晶的成核促进和生长抑制活性。在相同糖醛酸浓度下比较时,结石形成者的尿硫酸软骨素异构体和硫酸乙酰肝素均显示出增强尿液超滤液中内源性草酸钙晶体成核的能力,而只有正常对照受试者的尿硫酸乙酰肝素具有这种能力。4. 当进行类似测试时,组织来源的参考硫酸软骨素、硫酸皮肤素和肝素显示出可忽略不计的晶体成核促进活性。组织来源的硫酸乙酰肝素与尿硫酸乙酰肝素相似,显示出明显的晶体成核促进活性。5. 在所有研究的尿糖胺聚糖组分中均有明显的晶体生长抑制活性。特别是,正常对照受试者的尿硫酸乙酰肝素显示出比结石形成者或结石形成者和正常对照受试者的硫酸软骨素异构体更高的活性(P < 0.005)。

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