退变椎间盘聚集蛋白聚糖中硫酸软骨素链变化的实验研究

Comparative chondroitin salfate profiling from aggrecan of normal disc,internal disc disruptive and herniated discs in vitro

目的:研究内破裂及突出椎间盘细胞合成的聚集蛋白聚糖中硫酸软骨素链(CS)在长度及数量分布上的变化。方法:将正常椎间盘、内破裂(IDD)及突出椎间盘(LIDP)髓核或纤维环的组织20mg培养于24孔板中,用放射性同位素35S-硫酸盐和3H-丝氨酸标记新合成的蛋白聚糖分子。将聚集蛋白聚糖单体从培养的组织片中提取,用四氢硼酸钠或木瓜蛋白酶消化后,凝胶色谱分析硫酸软骨链的变化特征。结果:IDD椎间盘组织内细胞合成的聚集蛋白聚糖内CS链的数量明显减少,但长度保持相对正常;突出椎间盘组织中CS的数量和长度有明显下降和缩短,CS链数量的减少较IDD组织中更为严重。结论:IDD组织合成CS的减少主要是由于生成的CS链数量减少所致,而数量的进一步减少以及CS链长度的缩短是LIDP组织中CS合成量下降的主要原因。

Objective：To study the status of chondroitin salfate（CS） in discs of internal disc disruption（IDD） and lumbar intervertebral disc protrusion （LIDP）,which were compared with that of the normal discs in vitro an explant culture system.Method：20mg intervertebral disc tissues harvested from normal discs,IDD and LIDP donors were cultured in 24-well culture plate containing F12 medium,The newly synthesized proteoglycans were radiolabeled with aS-sulfate and 3H-serine.D1 monomer proteoglycans were then extracted from the tissue slices and digested with alkaline sodium borohydride or papain.Sieve chromatography were then performed to identify profile of CS chains.Result：The synthesis pattern was similar between NP and AF region.The aggrecan core protein in IDD contained fewer CS chains,but the chains were of normal length.The CS chain in LIDP was shortened,with a further decrease in the number of CS chains along the core protein compared with that of IDD.Conclusion：The decrease of CS content is mainly due to less number of the chains in IDD, and the decrease in number and lenghth of CS chain may result in the decrease of CS content in LIDP, which may play a role in generating discogenic.