Semiquantitative reverse transcription-polymerase chain reaction analysis of syndecan-1 and-4 messages in cartilage and cultured chondrocytes from osteoarthritic joints
Semiquantitative reverse transcription-polymerase chain reaction analysis of syndecan-1 and-4 messages in cartilage and cultured chondrocytes from osteoarthritic joints
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DOI：
10.1053/joca.1999.0286
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发表时间：
2000-01-01
影响因子：
7
通讯作者：
Pujol, JP
中科院分区：
文献类型：
--
作者：
Barre, PE;Redini, F;Pujol, JP
目的：确定软骨样品中的syndecan-1和syndecan-4的稳态（mRNA）水平的软骨样品和软骨细胞中衍生自人骨关节炎膝关节的稳态。通过半定量聚合酶链反应（PCR）测量 - 肌动蛋白）：RT-PCR允许检测在软骨样品和关节软骨细胞中，syndecan-1（首次）和Syndecan-4的syndecan-4。与正常外观的区域相比，软骨组织中syndecan-1的mRNA水平降低了，而Syndecan-4的mRNA组织显着增加。相比之下，源自纤元的骨关节炎软骨的培养软骨细胞中syndecan-1的表达比从完整软骨获得的细胞中的表达更高，而syndecan-4消息水平在两个地点之间没有差异。 - 在膝关节的骨关节炎降解过程中，Surface Syndecans 1和4被改变。二元联合基因表达可能与软骨细胞增殖和聚类有关，可能有助于软骨的混乱和OA过程的发展。隔离和培养软骨细胞，因为单层急剧改变了这些基因的表达，无法反映原位条件。 （c）2000年国际骨关节炎研究协会。
Objective: To determine the steady-state of messenger RNA (mRNA) levels of syndecan-1 and syndecan-4 in cartilage samples and chondrocytes derived from human osteoarthritic knee joints.Methods: Steady-state levels of gene-specific mRNA (relative to beta-actin) were measured-by semiquantitative polymerase chain reaction (PCR).Results:RT-PCR allowed detection of syndecan-1 (for the first time) and syndecan-4 in both cartilage samples and articular chondrocytes cultured as primary monolayers. The mRNA levels of syndecan-1 were reduced in cartilage tissue from heavily damaged compared to normal-looking areas whereas those of syndecan-4 were significantly increased. In contrast, the expression of syndecan-1 was higher in cultured chondrocytes derived from the fibrillated osteoarthritic cartilage than in cells obtained from intact cartilage, while the syndecan-4 message levels did not differ between the two sites.Conclusion: The expression of the cell-surface syndecans 1 and 4 is altered during the osteoarthritic degradative process of the knee joint. The discoordinate syndecan gene expression, which is probably related to the chondrocyte proliferation and clustering, may contribute to the disorganization of the cartilage and the development of OA processes; isolation and culturing the chondrocytes as monolayers dramatically change the expression of these genes and cannot reflect the in situ condition. (C) 2000 OsteoArthritis Research Society International.