Legendre F, Baugé C, Roche R, Saurel A S, Pujol J P
Laboratory of Connective Tissue Biochemistry, Faculty of Medicine, 14032 Caen Cedex, France.
Osteoarthritis Cartilage. 2008 Jan;16(1):105-14. doi: 10.1016/j.joca.2007.05.020. Epub 2007 Jul 12.
To determine the effect of avian chondroitin sulfate (CS) on interleukin-1beta (IL-1beta)-induced expression of genes related to catabolic, anabolic and inflammatory aspects in chondrocytes cultured in hypoxic alginate beads.
Articular chondrocytes from bovine metacarpal joint were isolated and cultured in alginate beads, using low oxygen atmosphere (5% O2). After 1-week exposure to CS (1, 10 and 100microg/ml), they were treated by recIL-1beta (10ng/ml) for 24 or 48h, in the presence of CS. RNA was extracted and used to determine, by quantitative reverse transcription-polymerase chain reaction, the steady-state levels of mRNAs encoding several genes related to anabolic, catabolic and inflammatory aspects. Glycosaminoglycan (GAG) synthesis was also assayed by 35S-sulfate incorporation.
CS decreased IL-1beta-induced expression of matrix metalloproteases-1, -3 and -13 and aggrecanases-1 and -2. It slightly enhanced the aggrecan core protein mRNA and the GAG synthesis. Inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) mRNA levels were found to be reduced by CS treatment. However, no CS-induced decrease of NO was observed in IL-1beta-treated chondrocytes, whereas prostaglandin E2 production was diminished in correlation with the COX-2 mRNA amounts. Furthermore, CS was capable of counteracting IL-1beta-depressed expression of transforming growth factor-beta (TGF-beta) receptors.
CS can repress expression of genes encoding proteolytic enzymes involved in cartilage degradation. It also inhibits IL-1beta-induced expression of the pro-inflammatory genes iNOS and COX-2 and restores TGF-beta receptors I and II (TGF-betaRI and RII) mRNA levels. These data suggest that CS may exert both chondroprotective and anti-inflammatory limited effects on articular cartilage that could have long-term beneficial action on the osteoarthritic process.
确定禽硫酸软骨素（CS）对在低氧海藻酸钠珠中培养的软骨细胞中白细胞介素-1β（IL-1β）诱导的与分解代谢、合成代谢和炎症方面相关基因表达的影响。
从牛掌指关节分离关节软骨细胞，在低氧气氛（5% O₂）下于海藻酸钠珠中培养。在暴露于CS（1、10和100μg/ml）1周后，在CS存在的情况下，用重组IL-1β（10ng/ml）处理24或48小时。提取RNA并通过定量逆转录-聚合酶链反应来测定编码与合成代谢、分解代谢和炎症方面相关的几个基因的mRNA的稳态水平。还通过³⁵S-硫酸盐掺入法测定糖胺聚糖（GAG）合成。
CS降低了IL-1β诱导的基质金属蛋白酶-1、-3和-13以及聚集蛋白聚糖酶-1和-2的表达。它略微增强了聚集蛋白聚糖核心蛋白mRNA和GAG合成。发现经CS处理后诱导型一氧化氮合酶（iNOS）和环氧化酶-2（COX-2）的mRNA水平降低。然而，在IL-1β处理的软骨细胞中未观察到CS诱导的NO减少，而前列腺素E₂的产生与COX-2 mRNA量相关而减少。此外，CS能够抵消IL-1β抑制的转化生长因子-β（TGF-β）受体的表达。
CS可抑制编码参与软骨降解的蛋白水解酶的基因的表达。它还抑制IL-1β诱导的促炎基因iNOS和COX-2的表达，并恢复转化生长因子-β受体I和II（TGF-βRI和RII）的mRNA水平。这些数据表明CS可能对关节软骨发挥软骨保护和抗炎有限作用，并可能对骨关节炎进程产生长期有益作用。
