Cole T C, Ghosh P, Taylor T K
Biochim Biophys Acta. 1986 Feb 19;880(2-3):209-19. doi: 10.1016/0304-4165(86)90082-6.
A group of young (2.0 +/- 0.6 years) (group 1) and old (9.7 +/- 1.5 years) (group 2) beagle dogs were given Na2 35SO4 (1.0 mCi/kg) intravenously 60 days prior to being killed to radiolabel their proteoglycans. Lumbar discs were removed and dissected into nucleus pulposus and annulus fibrosus. Proteoglycans were extracted at 4 degrees C from these tissues with buffered 4.0 M Gdn-HCl containing proteinase inhibitors, and purified by CsCl density gradient ultracentrifugation. The average hydrodynamic size and ability of the purified proteoglycans to aggregate in the presence of excess hyaluronic acid was determined by Sepharose CL-2B chromatography. The galactosamine/glucosamine ratios of these proteoglycans as well as their non-aggregating fractions were also ascertained. The proteoglycan content of discs of old animals was significantly less than in the young. The proportion of 35S-labelled, or non-labelled proteoglycans which could aggregate in the presence of hyaluronic acid was also much lower in the preparations isolated from the older discs. In contrast, the average hydrodynamic size of the non-aggregating proteoglycans isolated from the annuli fibrosi of group 2 animals were larger than the corresponding population of group 1 animals. Aminosugar analysis of these same proteoglycan fractions from older animals afforded galactosamine/glucosamine ratios (mean 1.81 +/- 0.14) which were less than the younger age group (mean 2.63 +/- 0.40). These data suggest that with ageing and degeneration the proteoglycans of the beagle disc undergo increased degradation with the accumulation in the annulus fibrosus of a population which is of larger average hydrodynamic size and richer in keratan sulphate than proteoglycans present in younger tissues.
一组年轻(2.0±0.6岁)(第1组)和老年(9.7±1.5岁)(第2组)的比格犬在处死前60天静脉注射Na2 35SO4(1.0 mCi/kg)以对其蛋白聚糖进行放射性标记。取出腰椎间盘并解剖为髓核和纤维环。在4℃下,用含有蛋白酶抑制剂的4.0 M Gdn-HCl缓冲液从这些组织中提取蛋白聚糖,并通过CsCl密度梯度超速离心进行纯化。通过琼脂糖CL-2B色谱法测定纯化的蛋白聚糖在过量透明质酸存在下的平均流体力学尺寸和聚集能力。还确定了这些蛋白聚糖及其非聚集部分的半乳糖胺/葡萄糖胺比率。老年动物椎间盘的蛋白聚糖含量明显低于年轻动物。从老年椎间盘分离的制剂中,在透明质酸存在下可聚集的35S标记或未标记的蛋白聚糖比例也低得多。相比之下,从第2组动物纤维环分离的非聚集蛋白聚糖的平均流体力学尺寸大于第1组动物的相应群体。对老年动物相同蛋白聚糖部分的氨基糖分析得出半乳糖胺/葡萄糖胺比率(平均1.81±0.14),低于年轻年龄组(平均2.63±0.40)。这些数据表明,随着衰老和退变,比格犬椎间盘的蛋白聚糖降解增加,在纤维环中积累了一群平均流体力学尺寸更大且硫酸角质素比年轻组织中存在的蛋白聚糖更丰富的蛋白聚糖。