类风湿关节炎患者造血干/祖细胞核因子κB1、κB2基因表达的研究

The expression of NFκB1 and NFκB2 genes in hematopoietic stem/progenitor cell of patients with rheumatoid arthritis

目的检测类风湿关节炎（rheumatoid arthritis,RA）患者外周血CD34＋造血干细胞/祖细胞（hematopoietic stem/progenitor cell,HSC/HPC）的核因子（nuclear factorκB,NFκB）κB1、κB2信号分子的基因表达水平,了解NFκB信号途径分子基因表达的异常与临床指标的关联性。方法收集24例RA患者,9例正常对照组,分别抽取50 ml抗凝外周血,淋巴细胞分离液分离单个核细胞,免疫磁珠分选CD34＋HSC/HPC,Trizol法提取总RNA,逆转录为cDNA,实时荧光定量聚合酶反应方法检测患者组和对照组NFκB1、NFκB2 mRNA表达水平的差异,并分析RA患者组甲氨蝶呤（Methotrexate,MTX）治疗组和未治疗组NFκB1、NFκB2mRNA表达水平变化,结合其它临床资料进行相关性分析。结果 RA患者组NFκB1 mRNA表达相对量高于正常对照组[6.57（1.08~76.71）,2.14（0.68~7.09）,P=0.013];NFκB2 mRNA表达与正常对照组无统计学差异。RA患者外周血CD34＋HSC/HPCNFκB1、NFκB2 mRNA表达与有无MTX的治疗无统计学差异,与疾病活动关节评分28（disease activity score 28,DAS28评分）、血沉、C反应蛋白等临床指标无相关性。结论 RA患者外周血CD34＋HSC/HPC NFκB1基因表达水平比正常对照增高,可能是RA患者CD34＋细胞重要缺陷。

This study aim to investigate the expression levels of NFκB1 and NFκB2 genes in hematopoietic stem/progenitor cell（HSC/HPC）derived from rheumatoid arthritis（RA）.We collected totally clinical date of 24 RA patients and 9 healthy controls.Peripheral blood mononuclear cells（PBMCs） derived from 50 ml of whole blood were separated by gradient centrifugation and CD34＋ cells were purified using an MACS magnetic separation device.Total RNA of CD34＋ cell was extracted and transcripted into cDNA.SYBR Green I dye based real-time quantitative PCR method was used to compare the expression of NFκB1 and NFκB2 in patients and controls,then the correlation with clinical data was analyzed.The expression levels of NFκB1 were [2.14（0.68-7.09）] and [6.57（1.08-76.71）] in normal controls and RA patients respectively（P = 0.013）.NFκB2 mRNA expression was no significant difference between normal controls and RA patients.However,the expression of NFκB1 and NFκB2 mRNA was no significant difference in CD34 ＋ HSC /HPC RA patients with or without methotrexate（MTX） treatment,and were not correlated with DAS28,erythrocyte sedimentation rate（ESR）,C reactive protein（CRP）,etc.The results suggest that the expression of NFκB1 gene in HSC/HPC of RA patients is higher than that from normal controls,which may be an important defect in CD34＋ HSC/HPC RA patients.