Horton W E, Udo I, Precht P, Balakir R, Hasty K
Laboratory of Biological Chemistry, Gerontology Research Center, National Institute on Aging, Baltimore, Maryland 21224, USA.
In Vitro Cell Dev Biol Anim. 1998 May;34(5):378-84. doi: 10.1007/s11626-998-0019-8.
The objective of this study was to determine if an immortalized mammalian chondrocyte cell line had a profile of matrix metalloproteinase (MMP) expression that was consistent with what has been reported for primary chondrocytes in vitro and in vivo. A combination of zymography, Western, and Northern analysis was used to examine the expression of MMPs that are relevant to cartilage degradation. Both interleukin-1beta and tumor necrosis factor alpha induced a 4- to 9-fold increase in the level of MMP-9 expression in conditioned media, and a 17- to 24-fold increase in MMP-3 mRNA. Other compounds such as basic fibroblast growth factor and staurosporine each increased MMP-9 expression individually and potentiated the effects of the two cytokines. Transforming growth factor beta had no positive or inhibitory effects. N-methyl arginine blocked the increase in nitric oxide observed following treatment with the cytokines but did not prevent the increased expression of MMPs. The pattern of metalloproteinase expression observed in IRC cells and the response to cytokines is very similar to what has been reported during the pathogenesis of osteoarthritis. The IRC cells should be useful as a model system to study basic mechanisms controlling chondrocyte MMP expression and to identify pharmacological modulators of this process.
本研究的目的是确定一种永生化的哺乳动物软骨细胞系是否具有与体外和体内原代软骨细胞所报道的基质金属蛋白酶（MMP）表达谱一致的表达谱。采用酶谱分析、蛋白质免疫印迹法和Northern分析相结合的方法，检测与软骨降解相关的MMP的表达。白细胞介素-1β和肿瘤坏死因子α均可使条件培养基中MMP-9表达水平增加4至9倍，使MMP-3 mRNA增加17至24倍。其他化合物如碱性成纤维细胞生长因子和星形孢菌素各自单独增加MMP-9表达，并增强两种细胞因子的作用。转化生长因子β没有正向或抑制作用。N-甲基精氨酸可阻断细胞因子处理后观察到的一氧化氮增加，但不能阻止MMP表达的增加。在IRC细胞中观察到的金属蛋白酶表达模式及对细胞因子的反应与骨关节炎发病机制中所报道的非常相似。IRC细胞作为研究控制软骨细胞MMP表达的基本机制以及鉴定该过程的药理学调节剂的模型系统应该是有用的。
