骨关节炎患者关节软骨TIMP-3启动子区甲基化水平的初步研究  

Investigation of TIMP-3 gene promoter methylation level in joint cartilage of osteoarthriti patients

作　　者：彭红春[1] 谢明[1] 沈奕[2] 彭丹[2] 

机构地区：[1]长沙市中心医院,长沙410001 [2]中南大学湘雅二医院,长沙410001

出　　处：《中国当代医药》2014年第2期4-7,共4页China Modern Medicine

基　　金：湖南省科技计划一般项目(2010FJ6004)

摘　　要：目的探讨关节软骨中组织金属蛋白酶抑制因子-3(TIMP-3)基因启动子区甲基化与其蛋白表达的相关性,并分析TIMP-3基因CpG岛异常甲基化与骨关节炎(OA)的关联性。方法应用甲基化特异性的聚合酶链反应(MSP)技术和免疫组织化学SP法分别检测14例健康人的正常关节软骨细胞和35例骨关节炎(OA)患者软骨细胞TIMP-3基因启动子区甲基化和蛋白表达情况。结果 OA患者和健康人关节软骨中TIMP-3基因启动子区均有甲基化修饰,其阳性率分别为74.3%(26/35)和35.7%(5/14),OA组TIMP-3基因启动子区甲基化率明显高于健康组(P<0.05)。14例健康人中,软骨细胞TIMP-3蛋白表达阳性10例(71.4%),而35例OA患者中,软骨细胞TIMP-3蛋白表达阳性11例(31.4%)。24例OA患者软骨细胞蛋白表达阴性的标本中,TIMP-3启动子区甲基化阳性21例(87.5%);26例TIMP-3启动子区甲基化阳性的标本中,蛋白表达阴性21例(80.8%),TIMP-3启动子区甲基化与蛋白表达呈显著负相关(P<0.05)。结论启动子区CpG岛高甲基化是OA患者关节软骨细胞TIMP-3表达失活的主要机制之一,其可能参与了OA的发生和发展。Objective To investigate the relationship between the methylation of tissue inhibitor of metalloproteinase-3 （TIMP-3） gene promoter and its protein expression, to study the potential role of hyper-methylation of TIMP-3 gene in the development of osteoarthritis. Methods The CpG islands methylation levels of TIMP-3 promoter in joint cartilage cells were studied by methylation-specific PCR（MSP）. TIMP-3 protein expression in joint cartilage cells were detected by immunohistochemical method in 35 osteoarthritis （OA） patients and 14 control health people. Results The positive percent of TIMP-3 promoter methylation levels in healthy people and OA patients were 35.7%（5/14） and 74.3%（26/35） respectively. The positive percent of TIMP-3 protein expression in OA patients were higher than healthy people （P〈 0.05）; In cartilage cells, the positive percent of TIMP-3 protein expression in healthy people and OA patients were 71.4%（10/14） and 31.4%（11/35） respectively. In 24 cases of OA samples with negative cartilage cells protein expres- sion, 21 cases （87.5%）was found with positive TIMP-3 promoter methylation, and in 26 cases of patients with positive TIMP-3 promoter methylation, protein expression was negative in 21 cases （80.8%）;Promoter methylation and TIMP-3 protein expression level showed significantly negative correlation. Conclusion TIMP-3 promoter is hyper-methlated in joint cartilage cells of OA patients. The hyper-methylation affects the protein expression of TIMP-3, which may play a crtical role in the occurrence and development of OA.

关 键 词：骨关节炎 甲基化 TIMP-3启动子 CPG岛 

分 类 号：R684.3[医药卫生—骨科学]