Hyaluronidase expression in cultured growth plate chondrocytes during differentiation
Hyaluronidase expression in cultured growth plate chondrocytes during differentiation
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DOI：
10.1007/s00441-004-0966-7
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发表时间：
2004-11-01
影响因子：
3.6
通讯作者：
Tanne, K
中科院分区：
文献类型：
--
作者：
Tanimoto, K;Suzuki, A;Tanne, K
透明质酸（HA）是软骨细胞外基质的主要成分，有助于其结构和功能的完整性，并在软骨细胞的分化中具有各种重要作用。HA代谢受合成代谢和分解代谢过程的调节;然而，细节尚未阐明。本研究的目的是阐明软骨细胞分化过程中透明质酸酶（HAase）mRNA（来自相关HAase基因：HYAL）和HAase活性的表达模式。从4周龄雄性日本兔的肋骨分离软骨组织和生长板软骨细胞。原位杂交分析软骨中HYAL mRNA的表达。通过实时定量聚合酶链反应分析，分析了每个软骨细胞分化阶段培养物中HYAL mRNA的表达水平。通过使用HA酶谱和酶联免疫吸附试验，从文化的条件培养基中的酶活性进行了检查。HYAL 1和HYAL 2 mRNA的表达水平在早期基质形成阶段分别增加约2.8倍和3.2倍，在肥大阶段分别增加约3.2倍和2.0倍。HYAL 3 mRNA在整个实验期间未检测到。HA酶活性在基质形成早期和肥大期增强。这些结果表明，选择性表达HYAL是必不可少的细胞外HA代谢过程中软骨细胞分化。
Hyaluronan (HA) is a major component of the extracellular matrix of cartilage, contributes to its structural and functional integrity, and has various important roles in the differentiation of chondrocytes. HA metabolism is regulated by both anabolic and catabolic processes; however, the details have not yet been clarified. The purpose of this study was to clarify the expression patterns of hyaluronidase (HAase) mRNAs (from the relevant HAase genes: the HYALs) and HAase activity during chondrocyte differentiation. Cartilage tissue and growth plate chondrocytes were isolated from the ribs of 4-week-old male Japanese rabbits. The expression of HYAL mRNAs in cartilage was analyzed by in situ hybridization. The expression levels of HYAL mRNAs in the culture were analyzed for each of the chondrocyte differentiation stages by means of quantitative real-time polymerase chain reaction analysis. Enzymatic activity in the conditioned medium from the cultures was examined by using HA zymography and an enzyme-linked immunosorbent-like assay. The expression levels of HYAL1 and HYAL2 mRNAs were enhanced about 2.8-fold and 3.2-fold at the maximum during the early matrix forming stage, respectively, and by about 3.2-fold and 2.0-fold at the maximum in the hypertrophic stage, respectively. HYAL3 mRNA was not detected throughout the experimental period. HAase activity was enhanced at the early matrix forming and hypertrophic stages. These results suggest that selective expression of HYALs is essential for extracellular HA metabolism during chondrocyte differentiation.