Arner E C, Kirkland J J
Cancer and Inflammatory Diseases Section, E.I. du Pont de Nemours and Company, Wilmington, DE 19880-0400.
Biochim Biophys Acta. 1989 Oct 13;993(1):100-7. doi: 10.1016/0304-4165(89)90148-7.
The effect of interleukin-1 (IL-1) on the size distribution of cartilage proteoglycans was studied using sedimentation field flow fractionation (SdFFF), a rapid, high-resolution technique for the separation of proteoglycan monomers and aggregates. During incubation of cartilage in control media, 35S-prelabeled proteoglycan was lost primarily from proteoglycan present in the monomer form; aggregates were conserved. In the presence of IL-1, both 35S-proteoglycan monomers and aggregates were lost, suggesting that IL-1 increases the susceptibility of aggregates to loss from the cartilage matrix. Evaluation of uronic acid as a measure of net change in proteoglycan content indicated that IL-1 causes a net decrease in both monomers and aggregates. Kinetic studies suggested that aggregates are degraded to monomers which then diffuse out of the matrix. Incorporation of [35S]sulfate into cartilage proteoglycans following exposure to IL-1 showed that synthesis of monomers and aggregates is inhibited similarly. SdFFF is a valuable technique for studying proteoglycan metabolism. With its use, changes in proteoglycan monomer and aggregate populations can be detected in response to cytokines such as IL-1.
利用沉降场流分级法(SdFFF)研究了白细胞介素-1(IL-1)对软骨蛋白聚糖大小分布的影响,沉降场流分级法是一种用于分离蛋白聚糖单体和聚集体的快速、高分辨率技术。在对照培养基中培养软骨的过程中,35S预标记的蛋白聚糖主要从以单体形式存在的蛋白聚糖中丢失;聚集体得以保留。在存在IL-1的情况下,35S蛋白聚糖单体和聚集体均丢失,这表明IL-1增加了聚集体从软骨基质中丢失的敏感性。以糖醛酸作为蛋白聚糖含量净变化的指标进行评估表明,IL-1导致单体和聚集体均出现净减少。动力学研究表明,聚集体降解为单体,然后扩散出基质。暴露于IL-1后,[35S]硫酸盐掺入软骨蛋白聚糖的情况表明,单体和聚集体的合成受到类似抑制。沉降场流分级法是研究蛋白聚糖代谢的一种有价值的技术。通过使用该技术,可以检测到蛋白聚糖单体和聚集体群体因诸如IL-1等细胞因子而发生的变化。