Kruse R, Merten M, Yoshida K, Schmidt A, Völker W, Buddecke E
Institut für Humangenetik der Universität, Bonn, FRG.
Basic Res Cardiol. 1996 Sep-Oct;91(5):344-52. doi: 10.1007/BF00788713.
Glycosaminoglycans are regular constituents of the arterial wall and essential for its structure and function. The arteriosclerosis-dependent changes of glycosaminoglycans were investigated, the degree of arteriosclerosis was monitored by the cholesterol content of the tissue. Histological characterization was achieved by electron microscopy. Total glycosaminoglycans were isolated from 33 delipidated segments of human aorta thoracica after exhaustive proteolytic digestion, and fractionated into the individual glycosaminoglycans by a multistep purification procedure. Chondroitin sulfate (CS), dermatan sulfate (DS), heparan sulfate (HS), and hyaluronate (HA) were identified and quantified by chemical and enzymatic analysis. The concentration of total and individual glycosaminoglycans, expressed as mg/g delipidated dry weight of tissue, decreased significantly with increasing cholesterol content of tissue (p = 0.0005-0.005). The extent of decrease differed between the individual glycosaminoglycans as indicated by a shift in the CS/DS:HA:HS ratio from 47:32:21 in low cholesterol aortic segments to 59:29:12 in cholesterol-rich specimens. Determination of the relative molecular masses (Mr) revealed 58 kDa for CS/DS and 92 kDa for HS with a (statistically not significant) increase of the molecular mass of CS/DS and a decrease of HS with increasing cholesterol content. The copolymeric CS/DS glycosaminoglycans were disintegrated enzymatically into CS and DS containing fragments. A significantly higher relative DS content (p = 0.01) was found in cholesterol-rich arterial tissue (32.5%) as compared with low cholesterol tissue samples (28.8%). Cell culture experiments revealed that human arterial HS is able to inhibit the proliferation of cultured human arterial smooth muscle cells. The HS concentration required for a 30% inhibition of smooth muscle cell proliferation was in the same order as the tissue concentration of HS. This confirms the function of HS as an endogenous inhibitor of cell division and its impact for the development of atherosclerosis.
糖胺聚糖是动脉壁的常规成分，对其结构和功能至关重要。研究了糖胺聚糖依赖于动脉硬化的变化，通过组织的胆固醇含量监测动脉硬化程度。通过电子显微镜进行组织学表征。在经过彻底的蛋白水解消化后，从33个脱脂的人胸主动脉段中分离出总糖胺聚糖，并通过多步纯化程序将其分离成各个糖胺聚糖。通过化学和酶促分析鉴定并定量硫酸软骨素（CS）、硫酸皮肤素（DS）、硫酸乙酰肝素（HS）和透明质酸（HA）。以mg/g脱脂组织干重表示的总糖胺聚糖和各个糖胺聚糖的浓度随着组织胆固醇含量的增加而显著降低（p = 0.0005 - 0.005）。如CS/DS:HA:HS比值从低胆固醇主动脉段的47:32:21转变为富含胆固醇标本中的59:29:12所示，各个糖胺聚糖的降低程度有所不同。相对分子质量（Mr）的测定显示CS/DS为58 kDa，HS为92 kDa，随着胆固醇含量的增加，CS/DS的分子量有（统计学上无显著意义的）增加，而HS的分子量降低。共聚物CS/DS糖胺聚糖被酶解成含有CS和DS的片段。与低胆固醇组织样本（28.8%）相比，在富含胆固醇的动脉组织（32.5%）中发现相对DS含量显著更高（p = 0.01）。细胞培养实验表明，人动脉HS能够抑制培养的人动脉平滑肌细胞的增殖。抑制平滑肌细胞增殖30%所需的HS浓度与组织中HS的浓度处于同一水平。这证实了HS作为细胞分裂内源性抑制剂的功能及其对动脉粥样硬化发展的影响。
