退变关节软骨细胞外基质降解酶DNA甲基化研究  

DNA methylation status of matrix - degrading enzymes in degenerating cartilage

作　　者：王瑞[1] 曹永平[1] 刘恒[1] 崔云鹏[1] 杨昕[1] 文立成[1] 柴卫兵[1] 孟志超[1] 吴浩[1] 潘利平[1] 周星彤[1] 李翔[1] 

机构地区：[1]北京大学第一医院骨科,100034

出　　处：《中华实验外科杂志》2016年第1期147-150,共4页Chinese Journal of Experimental Surgery

摘　　要：目的探讨不同退变程度关节软骨细胞外基质降解酶DNA甲基化水平。方法收集因退行性骨关节病（OA）行膝关节置换术患者的关节软骨40例。根据Outerbridge分级对所取软骨组织行组织学染色。检测不同退变程度软骨组织中基质金属蛋白酶（MMPs），如MMP-3、MMP-9、MMP-13以及蛋白聚糖酶（ADAM），如ADAMTS-4基因转录，并对以上基因启动子区域CpG位点甲基化进行评估。结果组织学检测显示，随着软骨退变加重，软骨细胞外蛋白聚糖丢失逐渐增多。Real-time PCR结果显示OA软骨组织中MMP-3、MMP-9、MMP-13基因转录均较正常软骨组织明显增加（1．845±0．280比0．391±0．180；1．414±0.222比0．498±0．127；1．532±0．158比0．637±0．184；P〈0．01），而基因启动子区域甲基化的CpG位点较正常软骨组织减少。结论OA软骨细胞外基质降解酶DNA启动子甲基化水平降低，可能引起基因转录增加及软骨细胞退变。Objective To evaluate the DNA methylation status of matrix - degrading enzymes in different degenerating cartilage. Methods 40 cartilage samples of the knee joint were collected during total knee arthroplasty surgery because of osteoarthritis （OA） from July 2013 to February 2014 in Peking U- niversity First Hospital. Cartilage tissues grouped by outerbridge classification were stained with Safranin O to identify proteoglycans loss. RNA was extracted for the assessment of mRNA expression of matrix metallo- proteinases （MMPs, such as MMP - 3, MMP - 9, MMP - 13 ） and aggrencanases （ ADAM, such as ADAMTS -4） from outerbridge 0 cartilage and outerbridge Ⅲ cartilage, respectively. DNA was extracted for the analysis of DNA promoter methylation status of these enzymes. Results The proteoglycans loss increased with the aggravation of cartilage degeneration. The expression of MMP - 3, MMP - 9 and MMP - 13 were increased significantly in OA cartilage compared with normal cartilage （ 1. 846 _± 0. 280 vs. 0. 391 ± 0. 180, 1.414±0.222 vs. 0.498 ±0. 127, 1.632 ±0. 158 vs. 0.637 ±0. 184, respectively; P〈0.01）. The amount of methylation CpG sites of these genes promotors in OA cartilage decreased compared with normal cartilage. Conclusion Decreased methylation status of the matrix - degrading enzymes genes may contribute to the increase in these enzymes production in OA cartilage.

关 键 词：骨关节炎 细胞外基质降解酶 DNA甲基化 

分 类 号：R692[医药卫生—泌尿科学]