Honda A, Abe M, Murota S, Mori Y
J Biochem. 1979 Feb;85(2):519-28. doi: 10.1093/oxfordjournals.jbchem.a132359.
The amount of glycosaminoglycan (GAG) in dry costal cartilage tissue of rats decreased with aging, while the GAG content in mg DNA (unit cartilage cell) remained the same with aging. These results can be explained by the finding that the total number of cartilage cells decreased with aging. Electrophoretic analysis showed that chondroitin 4-sulfate was the major GAG in rat costal cartilage of various ages. Rat costal cartilage of different ages was incubated with radioactive precursors, and newly synthesized GAG was prepared and the radioactivity analyzed to determine the biosynthetic activity. As to changes in the radioactivity uptake with aging per mg dry cartilage tissue, aging influenced [35S]sulfate incorporation into GAG more significantly than [3H]glucosamine incorporation into GAG. There was a significant decrease in the specific radioactivity of [35S]sulfate per mg DNA (unit cartilage cell), whereas the specific radioactivity of [3H]glucosamine per mg DNA did not change significantly with aging. Both the total sulfotransferase activity and the specific activity per mg DNA decreased significantly with aging. Analysis of disaccharide units formed after chondroitinase ABC digestion of labeled GAG isolated from young and old cartilage showed that the percentage of incorporation of [3H]glucosamine into deltaDi-OS increased significantly with aging. These results suggested that the appearance of nonsulfated positions in the structure of the chondroitin sulfate chain increased with aging. On the basis of gel chromatography on Bio-Gel A-1.5 m no significant difference in the approximate molecular size of chondroitin sulfate was observed between the young and old GAG samples. The present study indicated that the sulfation of chondroitin sulfate chains from rat costal cartilage decreased with the process of aging.
大鼠肋软骨组织干重中的糖胺聚糖(GAG)含量随衰老而降低,而每毫克DNA(单位软骨细胞)中的GAG含量在衰老过程中保持不变。这些结果可以通过软骨细胞总数随衰老而减少这一发现来解释。电泳分析表明,硫酸软骨素4-硫酸盐是不同年龄段大鼠肋软骨中的主要GAG。将不同年龄段的大鼠肋软骨与放射性前体一起孵育,制备新合成的GAG并分析其放射性以确定生物合成活性。至于每毫克干软骨组织随衰老的放射性摄取变化,衰老对[35S]硫酸盐掺入GAG的影响比对[3H]葡糖胺掺入GAG的影响更显著。每毫克DNA(单位软骨细胞)中[35S]硫酸盐的比放射性显著降低,而每毫克DNA中[3H]葡糖胺的比放射性随衰老没有显著变化。总硫酸转移酶活性和每毫克DNA的比活性均随衰老而显著降低。对从年轻和老年软骨中分离出的标记GAG进行软骨素酶ABC消化后形成的二糖单位分析表明,[3H]葡糖胺掺入δDi-OS的百分比随衰老显著增加。这些结果表明,硫酸软骨素链结构中未硫酸化位置的出现随衰老而增加。基于在Bio-Gel A-1.5m上的凝胶色谱分析,在年轻和老年GAG样品之间未观察到硫酸软骨素近似分子大小的显著差异。本研究表明,大鼠肋软骨硫酸软骨素链的硫酸化随衰老过程而降低。