Robinson D, Mirovsky Y, Halperin N, Evron Z, Nevo Z
Department of Clinical Biochemistry, Sackler School of Medicine, Tel Aviv University, Ramat Aviv, Israel.
Spine (Phila Pa 1976). 1998 Apr 15;23(8):849-55; discussion 856. doi: 10.1097/00007632-199804150-00001.
Characterization of the analytic profile of proteoglycans in the intervertebral discs at L4-L5 of nondiabetic (n = 5) and diabetic (n = 5) age-matched subjects. The discs used were discarded material from operations.
To clarify the reason for the higher risk of disc prolapse in diabetic patients.
The pathogenesis of diabetes results from a combination of neurologic dysfunctions and a yet undefined metabolic failure, which leads to an abnormal proteoglycan profile.
The following methods were used to determine the proteoglycan profile: the measurement of 35S-sulfate uptake per gram wet tissue into sulfated glycosaminoglycan using fresh tissue explants; extraction of proteoglycans by 4 M guanidinium chloride containing protease inhibitors, with further purification by ultracentrifugation on cesium chloride buoyant density gradient under dissociative conditions; total uronic acid and protein contents in the various gradient fractions; assessing the length of sugar side chains of isolated 35Sulfate-glycosaminoglycan molecules by separation of the glycosaminoglycan molecules on a Sepharose 6B-CL column; and paper chromatography of the final digest products of glycosaminoglycan molecules obtained by chondroitinase ABC, a glycosaminoglycan-degrading enzyme.
The findings show that discs from normal nondiabetic subjects have 15 times the rate of 35Sulfate incorporation into glycosaminoglycan molecules than do discs of diabetic patients. The proteoglycans of diabetic patients are banded at a lower buoyant density, indicating a lowered glycosylation rate and a lower number of sugar side chains per core protein. In discs of diabetic patients, there is a slight increase in the chain length of chondroitin sulfate. Further analysis of the glycosaminoglycan chains showed a decreased amount of keratan sulfate, compared with that in nondiabetic subjects. However, the total uronic acid content of the disc tissues and the ratio of uronic acid to protein of each fraction were unchanged in diabetic patients versus that in control subjects.
Discs in patients with diabetes have proteoglycans with lower buoyant density and substantially undersulfated glycosaminoglycan, which with the specific neurologic damage in these patients, might lead to increased susceptibility to disc prolapse.
对年龄匹配的非糖尿病患者（n = 5）和糖尿病患者（n = 5）L4 - L5椎间盘蛋白聚糖分析图谱进行特征描述。所用椎间盘为手术废弃材料。
阐明糖尿病患者椎间盘突出风险较高的原因。
糖尿病的发病机制是神经功能障碍和尚未明确的代谢衰竭共同作用的结果，这会导致蛋白聚糖图谱异常。
采用以下方法确定蛋白聚糖图谱：使用新鲜组织外植体测量每克湿组织中35S - 硫酸盐摄取到硫酸化糖胺聚糖中的量；用含蛋白酶抑制剂的4M氯化胍提取蛋白聚糖，在解离条件下通过氯化铯浮力密度梯度超速离心进一步纯化；各梯度级分中的总糖醛酸和蛋白质含量；通过在琼脂糖6B - CL柱上分离糖胺聚糖分子来评估分离出的35S - 硫酸化糖胺聚糖分子糖侧链的长度；以及对由糖胺聚糖降解酶软骨素酶ABC获得的糖胺聚糖分子最终消化产物进行纸层析。
研究结果表明，正常非糖尿病受试者的椎间盘将35S - 硫酸盐掺入糖胺聚糖分子的速率是糖尿病患者椎间盘的15倍。糖尿病患者的蛋白聚糖在较低的浮力密度处形成条带，表明糖基化速率降低且每个核心蛋白的糖侧链数量减少。在糖尿病患者的椎间盘中，硫酸软骨素的链长度略有增加。与非糖尿病受试者相比，对糖胺聚糖链的进一步分析显示硫酸角质素的量减少。然而，糖尿病患者椎间盘组织的总糖醛酸含量以及各组分中糖醛酸与蛋白质的比例与对照组相比没有变化。
糖尿病患者的椎间盘中蛋白聚糖的浮力密度较低且硫酸化程度明显不足，再加上这些患者存在特定的神经损伤，可能导致椎间盘突出的易感性增加。
