Lee J H, Fitzgerald J B, DiMicco M A, Cheng D M, Flannery C R, Sandy J D, Plaas A H, Grodzinsky A J
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Arch Biochem Biophys. 2009 Sep;489(1-2):118-26. doi: 10.1016/j.abb.2009.07.006. Epub 2009 Jul 14.
We studied changes in chondrocyte gene expression, aggrecan degradation, and aggrecanase production and activity in normal and mechanically injured cartilage co-cultured with joint capsule tissue. Chondrocyte expression of 21 genes was measured at 1, 2, 4, 6, 12, and 24h after treatment; clustering analysis enabled identification of co-expression profiles. Aggrecan fragments retained in cartilage and released to medium and loss of cartilage sGAG were quantified. Increased expression of MMP-13 and ADAMTS4 clustered with effects of co-culture, while increased expression of ADAMTS5, MMP-3, TGF-beta, c-fos, c-jun clustered with cartilage injury. ADAMTS5 protein within cartilage (immunohistochemistry) increased following injury and with co-culture. Cartilage sGAG decreased over 16-days, most severely following injury plus co-culture. Cartilage aggrecan was cleaved at aggrecanase sites in the interglobular and C-terminal domains, resulting in loss of the G3 domain, especially after injury plus co-culture. Together, these results support the hypothesis that interactions between injured cartilage and other joint tissues are important in matrix catabolism after joint injury.
我们研究了与关节囊组织共培养的正常和机械损伤软骨中软骨细胞基因表达、聚集蛋白聚糖降解、聚集蛋白聚糖酶产生及活性的变化。在处理后1、2、4、6、12和24小时测量21种基因的软骨细胞表达；聚类分析能够识别共表达谱。对保留在软骨中并释放到培养基中的聚集蛋白聚糖片段以及软骨硫酸化糖胺聚糖（sGAG）的损失进行了定量。MMP - 13和ADAMTS4的表达增加与共培养的影响聚类，而ADAMTS5、MMP - 3、TGF -β、c - fos、c - jun的表达增加与软骨损伤聚类。损伤后以及与关节囊组织共培养后，软骨内ADAMTS5蛋白（免疫组织化学）增加。软骨sGAG在16天内减少，损伤加共培养后最为严重。软骨聚集蛋白聚糖在球状间和C末端结构域的聚集蛋白聚糖酶位点处被切割，导致G3结构域丢失，尤其是在损伤加共培养后。总之，这些结果支持了以下假设：损伤软骨与其他关节组织之间的相互作用在关节损伤后的基质分解代谢中很重要。
