Gene expression and activity of cartilage degrading glycosidases in human rheumatoid arthritis and osteoarthritis synovial fibroblasts.
Gene expression and activity of cartilage degrading glycosidases in human rheumatoid arthritis and osteoarthritis synovial fibroblasts.
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DOI：
10.1186/ar2697
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发表时间：
2009
影响因子：
4.9
通讯作者：
Buzas EI
中科院分区：
文献类型：
--
作者：
Pásztói M;Nagy G;Géher P;Lakatos T;Tóth K;Wellinger K;Pócza P;György B;Holub MC;Kittel A;Pálóczy K;Mazán M;Nyirkos P;Falus A;Buzas EI
与基质金属蛋白酶类似，糖苷酶也在软骨降解中起主要作用。由这些酶产生的碳水化合物裂解产物在关节炎期间从降解软骨中释放。一些裂解产物（如透明质酸寡糖）已被证明与Toll样受体结合并提供内源性危险信号，而其他裂解产物（如N-乙酰葡糖胺）据报道具有软骨保护功能。在本研究中，我们首次系统地研究了关节内糖苷酶的表达。采用实时荧光定量PCR法检测类风湿关节炎和骨关节炎患者滑膜成纤维细胞和滑膜标本中β-D-氨基己糖苷酶、β-D-葡萄糖醛酸酶、透明质酸酶、精子粘附分子1和klotho基因的表达。使用显色或荧光底物表征β-D-葡糖醛酸酶、β-D-氨基葡萄糖苷酶和β-D-氨基半乳糖苷酶活性。还测试了滑膜成纤维细胞衍生的微泡的糖苷酶活性。根据我们的数据，β-D-氨基己糖苷酶、β-D-葡萄糖醛酸酶、透明质酸酶和klotho在滑膜中表达。氨基己糖苷酶是关节内表达的主要糖苷酶，并且其主要由滑膜成纤维细胞产生。HexA亚基基因是编码氨基己糖苷酶α链和β链的两个基因之一，其特征是基因表达最强。其次是HexB亚基基因和β-D-葡萄糖醛酸酶基因的表达，而透明质酸酶-1基因和klotho基因在滑膜成纤维细胞和滑膜样品中的表达相当低。肿瘤生长因子-β1在类风湿关节炎和骨关节炎来源的滑膜成纤维细胞中显著下调糖苷酶表达。此外，促炎细胞因子（包括TNFα、IL-1β和IL-17）可中度下调软骨降解糖苷酶的表达。根据我们目前的数据，在类风湿关节炎和骨关节炎中，滑膜和滑膜成纤维细胞表达的糖苷酶受到某些局部表达的细胞因子的负调节。这并不排除如果与差异上调的蛋白酶协同作用以消耗软骨中的糖胺聚糖，则这些酶可能显著促进两种关节疾病中的软骨降解的可能性。
Similar to matrix metalloproteinases, glycosidases also play a major role in cartilage degradation. Carbohydrate cleavage products, generated by these latter enzymes, are released from degrading cartilage during arthritis. Some of the cleavage products (such as hyaluronate oligosaccharides) have been shown to bind to Toll-like receptors and provide endogenous danger signals, while others (like N-acetyl glucosamine) are reported to have chondroprotective functions. In the current study for the first time we systematically investigated the expression of glycosidases within the joints. Expressions of β-D-hexosaminidase, β-D-glucuronidase, hyaluronidase, sperm adhesion molecule 1 and klotho genes were measured in synovial fibroblasts and synovial membrane samples of patients with rheumatoid arthritis and osteoarthritis by real-time PCR. β-D-Glucuronidase, β-D-glucosaminidase and β-D-galactosaminidase activities were characterized using chromogenic or fluorogenic substrates. Synovial fibroblast-derived microvesicles were also tested for glycosidase activity. According to our data, β-D-hexosaminidase, β-D-glucuronidase, hyaluronidase, and klotho are expressed in the synovial membrane. Hexosaminidase is the major glycosidase expressed within the joints, and it is primarily produced by synovial fibroblasts. HexA subunit gene, one of the two genes encoding for the alpha or the beta chains of hexosaminidase, was characterized by the strongest gene expression. It was followed by the expression of HexB subunit gene and the β-D-glucuronidase gene, while the expression of hyaluronidase-1 gene and the klotho gene was rather low in both synovial fibroblasts and synovial membrane samples. Tumor growth factor-β1 profoundly downregulated glycosidase expression in both rheumatoid arthritis and osteoarthritis derived synovial fibroblasts. In addition, expression of cartilage-degrading glycosidases was moderately downregulated by proinflammatory cytokines including TNFα, IL-1β and IL-17. According to our present data, glycosidases expressed by synovial membranes and synovial fibroblasts are under negative regulation by some locally expressed cytokines both in rheumatoid arthritis and osteoarthritis. This does not exclude the possibility that these enzymes may contribute significantly to cartilage degradation in both joint diseases if acting in collaboration with the differentially upregulated proteases to deplete cartilage in glycosaminoglycans.
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影响因子：
--
作者：
Li, Rachel W.;Freeman, Craig;Smith, Paul N.
通讯作者：
Smith, Paul N.
影响因子：
4.8
作者：
Bunbury, Allyson;Potolicchio, Ilaria;Santambrogio, Laura
通讯作者：
Santambrogio, Laura
影响因子：
4.9
作者：
Kanangat, Siva;Postlethwaite, Arnold;Schaberg, Dennis
通讯作者：
Schaberg, Dennis
影响因子：
7
作者：
Fuchs, S;Skwara, A;Dankbar, B
通讯作者：
Dankbar, B
影响因子：
4.9
作者：
Jones GC;Riley GP
通讯作者：
Riley GP