骨关节炎患者关节软骨细胞合成和分解基因的表达研究

Study on the Expression of Synthesis and Decomposition Genes of Articular Cartilage Cells in Patients with Osteoarthritis

目的:初步探讨合成和分解相关基因Ⅱ型胶原a1(COL2A1)、聚蛋白多糖(ACAN)、含Ⅰ型血小板结合蛋白基序结构的去整合素金属蛋白酶-5(ADAMTs-5)、基质金属蛋白酶组织抑制剂-3(TIMP-3)在骨关节炎患者关节软骨细胞和外周血的表达水平。方法:入选行关节置换术的骨关节炎患者3例、类风湿关节炎患者3例和创伤截肢患者4例,分成骨关节炎组、类风湿关节炎组和创伤对照组,分别取离体关节软骨标本体外培养软骨细胞;入选骨关节炎患者9例、类风湿关节炎患者10例和健康对照者10例,分成骨关节炎组、类风湿关节炎组和正常对照组,分别抽取4 mL外周静脉血提取外周血单个核细胞,利用实时荧光定量聚合酶链反应法(RT-PCR)检测关节软骨细胞和外周血单个核细胞COL2A1、ACAN、ADAMTs-5、TIMP-3基因表达。采用非配对资料t检验比较各基因分别在关节软骨细胞、外周血单个核细胞表达的组间差异。结果:骨关节炎组与创伤对照组组间比较提示ACAN、TIMP-3、COL2A1基因在骨关节炎关节软骨细胞表达下调,ADAMTs-5基因表达上调,TIMP-3表达低于创伤对照组(P<0.05);各基因在骨关节炎外周血单个核细胞表达无统计学意义(P>0.05)。结论:TIMP-3、COL2A1、ACAN和ADAMTs-5基因在关节软骨细胞中的异常表达趋势对骨关节炎发生发展有预警作用,在外周血的表达无特异性。

Objective：To study the expression level of synthesis and decomposition genes such as COL2A1,ACAN,ADAMTs-5 and TIMP-3 in articular cartilage cells and peripheral blood.Methods：There were three selected groups：an osteoarthritis group（3 cases for joint replacement）,a rheumatoid arthritis group（3 cases with rheumatoid arthritis） and a trauma control group（4 cases of amputation due to trauma）.Cartilage specimens in vitro were taken to culture chondrocytes.In addition,Peripheral blood mononuclear cells were obtained from 4 mL peripheral vein blood taken from the other three groups：an osteoarthritis group（9 cases with osteoarthritis）,a rheumatoid arthritis group（10 cases with rheumatoid arthritis）and a health control group（10 healthy people）.RT-PCR method was used to detect the gene expressions of COL2A1,ACAN,ADAMTs-5 and TIMP-3 in articular cartilage cells and peripheral blood.Non-pairing data t was used to test and compare the expression differences of genes in articular chondrocytes and peripheral blood mononuclear cells.Results：The comparison between the osteoarthritis group and the trauma control group suggested that the expression of ACAN,TIMP-3 and COL2A1 had a down regulation in osteoarthritic articular cartilage cells,the expression of ADAMTS-5 was up-regulated,and the expression of TIMP-3 was lower than that of the trauma control group（P 〈0.05）.The expression of each gene in peripheral blood mononuclear cells was not statistically significant（P 〈0.05）.Conclusion：The abnormal expression of TIMP-3,COL2A1,ACAN and ADAMTs-5 in articular chondrocytes has a warning effect on the occurrence and development of osteoarthritis but no specificity in peripheral blood.