类风湿关节炎外周血PBMC中MIP-1α和RANTES的表达

The Expression of MIP-1α and RANTES in the Peripheral Blood Mononuclear Cells of Patients with Rheumatoid Arthritis

【目的】研究巨噬细胞炎症蛋白 1α(MIP 1α)和依赖激活正常T细胞表达分泌的调节蛋白(RANTES)在类风湿关节炎 (RA)患者外周血单个核细胞 (PBMC)中的mRNA水平的表达。探讨它们在RA发病机制中可能起的作用和与RA的疾病活动程度相关性。【方法】分别分离 32例RA、6例骨关节炎 (OA)以及 10例健康成人 (HC)的PBMC ,提取细胞总RNA ,行逆转录PCR ,将目的基因与内参照基因GAPDH共扩增。统计扩增的目的基因片段与内参照基因片段比值 ,并与RA患者疾病活动指标行相关性分析。【结果】RA、OA、HC各组PBMC标本MIP 1α与GAPDHRT PCR共扩增产物光密度比值与OA组、HC组比较均有显著性差异 ,P <0 .0 5。RA组PBMC标本RANTES与GAPDHRT PCR共扩增产物光密度比值与OA组、HC组比较无显著差异 ,P >0 .0 5。RA患者PBMC中MIP 1α和RANTESmRNA表达水平均与疾病活动性指标如病程、ESR、CRP、肿胀关节数、疼痛关节数、关节X线分期、RF滴度等未发现有相关性。【结论】趋化因子MIP 1α在RA患者PBMC中的mRNA表达水平高于OA和健康成年人 ,提示很可能在介导淋巴细胞从外周血进入炎症关节部位的过程中起着重要作用。

To study the mRNA expression levels of two intercellular (C-C) chemokines-macrophage inflammatory protein-1a ( MIP-1α) Released on Activation,Normal T-cell Expressed and Secreted(RANTES) in the peripheral blood mononuclear cells (PBMCs) isolated from patients with rheumatoid arthritis (RA) and explore their possible roles played in the pathogenesis of RA as well as their correlativity with the degree of disease activity. PBMCs were collected from 32 patients with RA, 6 patients with osteoarthritis (OA) and 10 healthy controls (HC). The total cellular RNAs were extracted and then reverse transcription-polymerase chain reaction (RT-PCR) was performed. The target gene and internal reference gene glyceraldehydes-3-phosphate dehydrogenase (GAPDH) were amplified together. The PCR products were gel-fractionated , stained with ethidium bromide and analyzed by densitometry. So the expression of MIP-1α and RANTES was semi-quantitated by MIP-1α/GAPDH or RANTES/GAPDH ratio.The OD ratio of MIP-1α co-amplified products in RA group was significantly increased as compared with those in OA HC group . With regard to the OD ratio of RANTES co-amplified products , there were no significant differences between RA and OA group or HC group . Furthermore, any correlation between the mRNA expression levels of MIP-1α/ RANTES in PBMCs of RA patients and criteria of disease activity [e.g. course of disease, erythrocyte sedimentation rate (ESR), C reactive protein (CRP), number of swollen or tender joints, X-ray grade of joints and rheumatoid factor (RF) titer etc.] was not found.[Conclusion]The mRNA expression level of C-C chemokine MIP-1α in PBMCs of RA patients is higher than that of OA patients or healthy adults. It suggests that MIP-1α probably plays an important role in attracting lymphocytes from peripheral blood into inflammatory joints and is involved in the procedure of local inflammation.