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目的寻找在 SLE 分子发病机制中起关键作用的致病基因的候选基因。方法通过高密度基因芯片技术,得到初发性 SLE 患者外周血单核细胞及 T、B 淋巴细胞亚群的基因表达谱,在差异表达显著的基因中挑选4个位于 SLE 易感区段的基因,通过 TaqMan 实时 PCR 技术在55例 SLE 患者中验证基因的差异表达。结果得到初发性 SLE 患者外周血单核细胞及 T、B 淋巴细胞亚群的基因表达谱,确定 TRIM,KLRC1,IL-4R 及 EGR-1基因为 SLE 易感基因的候选基因,实时 PCR 结果证明:SLE 患者外周血单核细胞中,TRIM 基因△Ct 值显著低于正常人对照(P<0.05),基因表达水平为正常对照的2.43倍,IL-4R 基因与 EGR1基因的△Ct 值显著高于正常人对照(P<0.05),基因表达水平分别为正常对照的0.33倍、0.26倍,KLRC1基因的△Ct 值显著高于正常人对照(P<0.01),基因表达水平为正常对照的0.56倍,在 SLE 患者外周血 T 细胞亚群中,TRIM 基因△Ct值显著低于正常人对照(P<0.01),基因表达水平为正常对照的2.94倍,IL4R 基因与 EGR1基因的△Ct 值显著高于正常人对照(P<0.01),基因表达水平分别为正常对照的0.32、0.21倍,LRC1基因的△Ct 值显著高于正常人对照(P<0.01),基因表达水平为正常对照的0.56倍。在 SLE 患者外周血 B 淋巴细胞亚群中,由于 B细胞淋巴亚群所占淋巴细胞比率过低,基因检测数据不稳定,故未作统计学分析。结论在系统性红斑狼疮的发病中,患者外周血单核细胞及 T、B 淋巴细胞亚群细胞基因表达水平发生了改变,主要集中于一系列免疫相关基因,其中 TRIM,KLRC1,IL-4R 及 EGR-1基因在患者群中变化显著,初步断定为 SLE 易感基因的候选基因。
Objective To find candidate genes of pathogenic genes that play key roles in the pathogenesis of SLE. Methods Gene expression profiles of peripheral blood mononuclear cells and T, B lymphocyte subsets in patients with primary SLE were obtained by high-density microarray. Four genes were identified in susceptible segments of SLE , Differential expression of the genes was verified in 55 SLE patients by TaqMan real-time PCR technique. Results The gene expression profiles of peripheral blood mononuclear cells and T lymphocyte subsets in patients with primary SLE were obtained. The candidate genes of TRIM, KLRC1, IL-4R and EGR-1 gene were identified as SLE susceptible genes. Real-time PCR results It was proved that the △ Ct value of TRIM gene in peripheral blood mononuclear cells of SLE patients was significantly lower than that of normal controls (P <0.05), the gene expression level was 2.43 times that of the normal controls, and the △ Ct value of IL-4R gene and EGR1 gene was significant (P <0.05). The gene expression level was 0.33-fold and 0.26-fold higher than that of the normal control respectively. The △ Ct value of KLRC1 gene was significantly higher than that of the normal human control (P <0.01) 0.56-fold. The △ Ct values of TRIM gene in peripheral blood T lymphocyte subsets of SLE patients were significantly lower than those in normal controls (P <0.01), and the gene expression levels were 2.94 times of the normal controls. The △ Ct values of IL4R and EGR1 (P <0.01). The gene expression levels were 0.32 and 0.21 times that of the normal control respectively. The △ Ct value of LRC1 gene was significantly higher than that of the normal control (P <0.01), and the gene expression level was significantly higher than that of the normal control 0.56 times. In SLE patients with peripheral blood B lymphocyte subsets, due to B lymphocyte subsets of lymphocyte ratio is too low, the genetic test data is not stable, so no statistical analysis. Conclusion In the pathogenesis of systemic lupus erythematosus, the gene expression of peripheral blood mononuclear cells and T, B lymphocyte subsets changed, mainly in a series of immune related genes, including TRIM, KLRC1, IL-4R and EGR-1 gene in patients with significant changes, initially identified as SLE susceptibility gene candidate gene.