Mechanical compression of cartilage explants induces multiple time-dependent gene expression patterns and involves intracellular calcium and cyclic AMP
Mechanical compression of cartilage explants induces multiple time-dependent gene expression patterns and involves intracellular calcium and cyclic AMP
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DOI：
10.1074/jbc.m400437200
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发表时间：
2004-05-07
影响因子：
4.8
通讯作者：
Grodzinsky, AJ
中科院分区：
文献类型：
--
作者：
Fitzgerald, JB;Jin, M;Grodzinsky, AJ
软骨细胞受到机械力的影响以重塑软骨细胞外基质。先前的研究已经证明了机械力对特定细胞外基质分子的生物合成和mRNA水平变化的影响，并且已经确定了可能涉及的某些信号通路。然而，基因转录的机械调控的广泛程度和动力学尚未深入研究。我们应用静态压缩应变牛软骨外植体1和24小时之间的时间，并测量了28个基因的反应，使用真实的时间PCR。在细胞内钙螯合剂或环AMP活化蛋白激酶A抑制剂存在下也进行压缩时间过程。聚类分析的数据揭示了四个主要的表达模式：两组包含瞬时上调或持续时间增强的表达谱，每个可以细分为基因，没有或不需要细胞内钙释放和环AMP激活的蛋白激酶A的机械调节。聚集蛋白聚糖、II型胶原和连接蛋白的转录水平在50%压缩的前8小时内上调了2 -3倍，随后下调至低于自由膨胀对照的水平。基质金属蛋白酶-3、-9和-13、聚集蛋白聚糖酶-1和基质蛋白酶调节剂环氧合酶-2的转录水平随着50%压缩持续时间的增加而增加2-16倍。因此，转录的蛋白质参与基质重塑和catastrophic占主导地位的合成代谢基质蛋白作为静态压缩的持续时间增加。即刻早期基因c-fos和c-jun在50%压缩的前8 h内分别显著上调6-30倍，并在24 h后保持上调。
Chondrocytes are influenced by mechanical forces to remodel cartilage extracellular matrix. Previous studies have demonstrated the effects of mechanical forces on changes in biosynthesis and mRNA levels of particular extracellular matrix molecules, and have identified certain signaling pathways that may be involved. However, the broad extent and kinetics of mechano-regulation of gene transcription has not been studied in depth. We applied static compressive strains to bovine cartilage explants for periods between 1 and 24 h and measured the response of 28 genes using real time PCR. Compression time courses were also performed in the presence of an intracellular calcium chelator or an inhibitor of cyclic AMP-activated protein kinase A. Cluster analysis of the data revealed four main expression patterns: two groups containing either transiently up-regulated or duration-enhanced expression profiles could each be subdivided into genes that did or did not require intracellular calcium release and cyclic AMP-activated protein kinase A for their mechano-regulation. Transcription levels for aggrecan, type II collagen, and link protein were up-regulated similar to2-3-fold during the first 8 h of 50% compression and subsequently down-regulated to levels below that of free-swelling controls by 24 h. Transcription levels of matrix metalloproteinases-3, -9, and -13, aggrecanase-1, and the matrix protease regulator cyclooxygenase-2 increased with the duration of 50% compression 2-16-fold by 24 h. Thus, transcription of proteins involved in matrix remodeling and catabolism dominated over anabolic matrix proteins as the duration of static compression increased. Immediate early genes c-fos and c-jun were dramatically up-regulated 6-30-fold, respectively, during the first 8 h of 50% compression and remained up-regulated after 24 h.