骨关节炎滑膜成纤维细胞与皮肤成纤维细胞基因表达的差异(英文)

Differences in gene expressions between synovioblast and skin fibroblast in patients with osteoarthritis

背景:近年来,发现一些基因的单核苷酸多态性可能与骨关节炎的易感性有关,以往研究主要集中于骨关节炎与类风湿性关节炎滑膜细胞基因表达差异,滑膜成纤维细胞与其他组织成纤维细胞基因表达差异与骨关节炎的关系有待研究。目的:观察骨关节炎滑膜成纤维细胞与皮肤成纤维细胞基因表达的差异。设计:观察对比分析。单位:深圳市人民医院。材料:骨关节炎患者(均符合1995年美国风湿病学会的骨关节炎诊断标准)滑膜及正常志愿者皮肤由深圳市人民医院骨科提供,所有提供皮肤者均对实验项目知情同意。RPMI 1640培养基、胎牛血清、TRIZOL试剂(美国Invitrogen Life Technologies公司),pGEM-T质粒系统(美国Progema公司),Display PROFILE-BASIC及Display PROFILEProbe试剂盒(美国Qbiogen公司)。方法:实验于2005-01/06在深圳市人民医院完成。选取骨关节炎患者滑膜,经原代培养获得滑膜成纤维细胞,采用正常志愿者原代培养皮肤成纤维细胞作为对照。采用限制性酶切片段差异展示技术分离骨关节炎患者滑膜成纤维细胞与皮肤成纤维细胞差异表达的基因,使用blast比对将所得序列与Genbank中所有序列进行比对分析。主要观察指标:关节炎患者滑膜成纤维细胞与皮肤成纤维细胞基因表达的差异。结果:骨关节炎患者滑膜成纤维细胞与正常志愿者皮肤成纤维细胞相比,超氧化物歧化酶、TFPI2、CXCL2、CXCL6、转化生长因子基因表达水平较高。结论:超氧化物歧化酶、TFPI2、CXCL2、CXCL6、转化生长因子基因在骨关节炎滑膜成纤维细胞表达水平高于皮肤成纤维细胞中相应基因表达水平,表明这些基因在骨关节炎疾病的发生过程中可能起到一定作用。

BACKGROUND： During recent years, mononucleotide polymorphism of some genes is possibly related to affectability of osteoarthritis （OA）. However, previous researches mainly compare the gene expression of synoviocytes between OA and rheumatoid arthritis （RhA）; therefore, the correlation of gene expression between synovioblast and fibroblast in other tissues should be further studied as compared with OA. OBJECTIVE ： To observe the differences of gene expression between OA synovioblast and skin fibroblast. DESIGN ： Observational contrast analysis. SETTING ： People＇s Hospital of Shenzhen City. PARTICIPANTS： Synovium tissue was derived from OA patients who received replacement of knee joint in the Department of Orthopaedics, People＇s Hospital of Shenzhen City. All OA patients met the diagnostic criteria of osteoarthritis established by American College of Rheumatology in 1995. Three patients including 1 male and 2 females aged more than 65 years old and they did not have cardiac and pulmonary disease and diabetes mellitus. Three male normal volunteers who aged 25 to 35 years did not have rheumatic disease, osteoarthritis and dermatosis. All subjects provided a confirmed consent. The main reagents were detailed as follows： RPMI1640 culture medium, fetal bovine serum and TRIZOL agent （Invitrogen Life Technologies Company, USA）; pGEM-T pUC （Progema Company, USA）; Display PROFILE-BASIC and Display PROFILE Probe kits （Qbiogen Company, USA）. METHODS ： The experiment was carried out in People＇s Hospital of Shenzhen City from January to June 2005. Synovium of OA patients were treated with primary culture to obtain synovioblast; meanwhile, skin fibroblast treated with primary culture from normal subjects was regarded as the control group. Restricted enzyme section differential display was used to separate the different-expressed genes of synovioblast and skin fibroblast in OA patients. In addition, blast technique was used to compare the resulted ranks with Genbank ranks. MAIN OUTCOME MEASURES ： Differences of gene expression between synovioblast and skin fibroblast in OA patients. RESULTS： Gene expressions of superoxide dismutase （SOD）, TFPI2, CXCL2, CXCL6 and transforming growth factor （TGF） were high in synovioblast of OA patients as compared with those in skin fibroblast of normal subjects. CONCLUSION： Gene expressions of SOD, TFPI2, CXCL2, CXCL6 and TGF are high in synovioblast of OA patientsaSof compared with those in skin fibroblast of normal subjects. This suggests that gene may play a certain role in onset of OA.