牛关节软骨外植体中蛋白聚糖转化动力学

J.W. Von Den Hoff , G.P.J. Van Kampen, R.J. Van De Stadt, J.K. Van Der Korst
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引用次数: 7

摘要

研究了成熟牛关节软骨外植体培养中蛋白聚糖的周转。本研究的目的是比较新合成的蛋白聚糖和内源性蛋白聚糖的体外周转率。为了确定稳态蛋白多糖代谢的条件,软骨在不同血清浓度下维持。在含20%胎牛血清的培养基中达到稳定状态。利用稳态培养中蛋白聚糖的合成速率和标记蛋白聚糖的半衰期来计算稳态下新合成蛋白聚糖代谢池的大小。这个代谢池被证明等于基质中蛋白多糖的总量。结果表明,基质中所有的蛋白聚糖在体外具有相同的半衰期。采用另一种方法,从软骨和用[35S]硫酸盐预标记的稳态培养培养基中分离聚合蛋白。比较了软骨聚集蛋白的糖胺聚糖与中等聚集蛋白的糖胺聚糖的比活性。这些在整个文化时期都是一样的。这说明新合成的聚集蛋白和内源性的聚集蛋白在体外具有相同的周转率。讨论了外植体培养系统对蛋白聚糖周转研究的意义。
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Kinetics of Proteoglycan Turnover in Bovine Articular Cartilage Explants

The turnover of proteoglycans was studied in explant cultures of mature bovine articular cartilage. The aim of the study was to compare the in vitro turnover rates of newly synthesized proteoglycans and endogenous proteoglycans. Cartilage was maintained in the presence of various serum concentrations in order to determine the conditions of steady-state proteoglycan metabolism. The steady state was achieved in medium containing 20% fetal calf serum.

The proteoglycan synthesis rate and the half-life of labeled proteoglycans in steady-state cultures were used to calculate the size of the metabolic pool of newly synthesized proteoglycans in steady state. This metabolic pool was shown to be equal to the total amount of proteoglycans in the matrix. It is concluded that all of the proteoglycans in the matrix have the same half-life in vitro.

Taking another approach, aggrecan was isolated from the cartilage and the medium of steadystate cultures prelabeled with [35S] sulfate. The specific activity of the glycosaminoglycans from cartilage aggrecan were compared with that of glycosaminoglycans from medium aggrecan. These proved to be the same throughout the culture period. This shows that newly synthesized aggrecan and endogenous aggrecan have the same turnover rate in vitro.

The significance of explant culture systems for the study of proteoglycan turnover is discussed.

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