Bishop P B, Pearce R H
Department of Pathology, Faculty of Medicine, University of British Columbia, Vancouver, Canada.
J Orthop Res. 1993 May;11(3):324-31. doi: 10.1002/jor.1100110303.
Lumbar spines collected postmortem were assigned to one of two groups: group 1--three spines with healthy discs, or group 2--three spines with severely degenerated discs. The proteoglycans (PGs) of the cartilaginous end-plate (CEP) were extracted with 4 M guanidinium chloride containing protease inhibitors and were purified by caesium chloride density gradient ultracentrifugation. On Sepharose CL-2B chromatography, the most dense 20% of the gradient (the A1 fraction) showed two subfractions, one eluting near the void volume and one partitioned by the gel. Both fractions resembled those of the nucleus pulposus and the anulus fibrosus in the number of components seen on agarose-polyacrylamide gel electrophoresis. Both fractions changed with ageing/degeneration; the ratio of keratan sulphate to chondroitin sulphate, which was about 1 in group 1, increased to about 3 in group 2; the hydrodynamic volumes fell; the electrophoretically distinguishable component of lowest mobility disappeared while new, highly mobile components appeared; and the water content decreased slightly. Clearly, the PGs of the CEP of degenerated intervertebral discs differed from those of healthy discs; this supports the view that the CEP participates in the process of ageing/degeneration in the disc.
第1组——三个椎间盘健康的腰椎；第2组——三个椎间盘严重退变的腰椎。用含蛋白酶抑制剂的4M氯化胍提取软骨终板（CEP）的蛋白聚糖（PGs），并通过氯化铯密度梯度超速离心进行纯化。在琼脂糖CL-2B柱层析上，梯度中最致密的20%（A1组分）显示出两个亚组分，一个在空体积附近洗脱，另一个被凝胶分配。在琼脂糖-聚丙烯酰胺凝胶电泳上观察到的组分数量方面，这两个组分都与髓核和纤维环的组分相似。这两个组分都随衰老/退变而变化；硫酸角质素与硫酸软骨素的比例在第1组中约为1，在第2组中增加到约3；流体动力学体积下降；电泳迁移率最低的可区分组分消失，同时出现了新的、高迁移率的组分；含水量略有下降。显然，退变椎间盘的CEP的PGs与健康椎间盘的不同；这支持了CEP参与椎间盘衰老/退变过程的观点。
