van der Kraan P M, de Vries B J, Vitters E L, van den Berg W B, van de Putte L B
Department of Rheumatology, St. Radboud Hospital, University of Nijmegen, The Netherlands.
J Orthop Res. 1989;7(5):645-53. doi: 10.1002/jor.1100070504.
We have studied the effect of environmental sulfate concentration on the glycosaminoglycan synthesis of anatomically intact patellar cartilage of the mouse in vitro. Incubation of mouse patellae in medium with sulfate concentrations below 0.5 mM resulted in a diminished incorporation of sulfate but in unaltered incorporation of glucosamine. This suggested the synthesis of undersulfated glycosaminoglycans under these conditions. We characterized glycosaminoglycans synthesized at three different sulfate concentrations: a sulfate concentration physiological for the mouse (1.0 mM), a sulfate concentration in the range where sulfate incorporation was strongly diminished (0.1 mM), and an extremely low sulfate concentration (10 nM). Analysis of glycosaminoglycan disaccharides and DEAE anion chromatography of the glycosaminoglycans could not confirm the synthesis of undersulfated glycosaminoglycans at 0.1 mM. The chromatogram of glycosaminoglycans synthesized in medium containing 10 nM showed the presence of a very low sulfated glycosaminoglycan pool not observed at higher medium sulfate concentrations. Intermediately sulfated glycosaminoglycans were also synthesized during incubation with 10 nM sulfate. So, our data indicate that only very low sulfate concentrations in the medium lead to the synthesis of undersulfated glycosaminoglycans and that the sulfation mechanism of murine patellar cartilage chondrocytes does not seem to fit completely in an "all-or-nothing" pattern.
我们研究了环境硫酸盐浓度对体外培养的小鼠解剖结构完整的髌软骨中糖胺聚糖合成的影响。将小鼠髌骨置于硫酸盐浓度低于0.5 mM的培养基中孵育,结果硫酸盐掺入减少,但葡萄糖胺掺入未改变。这表明在这些条件下合成了低硫酸化的糖胺聚糖。我们对在三种不同硫酸盐浓度下合成的糖胺聚糖进行了表征:一种对小鼠而言是生理浓度的硫酸盐(1.0 mM),一种硫酸盐掺入强烈减少范围内的浓度(0.1 mM),以及一种极低的硫酸盐浓度(10 nM)。对糖胺聚糖二糖的分析以及糖胺聚糖的DEAE阴离子色谱分析无法证实0.1 mM时低硫酸化糖胺聚糖的合成。在含有10 nM硫酸盐的培养基中合成的糖胺聚糖的色谱图显示存在一个在较高培养基硫酸盐浓度下未观察到的极低硫酸化糖胺聚糖池。在10 nM硫酸盐孵育期间也合成了中等硫酸化的糖胺聚糖。因此,我们的数据表明,只有培养基中极低的硫酸盐浓度会导致低硫酸化糖胺聚糖的合成,并且小鼠髌软骨软骨细胞的硫酸化机制似乎并不完全符合“全或无”模式。