目的检测系统性红斑狼疮(SLE)患者CD4+T细胞中miR326、E26转录因子-1(Ets-1)表达量及Th17细胞比例,分析三者之间的相关性及与狼疮活动度、系统受累情况的关联,探讨SLE可能的致病机制。方法选取符合2009年美国风湿病学会诊断标准的SLE患者40例,记录临床资料,选取无自身免疫性疾病的健康志愿者14例。流式细胞术检测Th17(CD4+IL-17A+)/CD4+T细胞比例,磁珠分选CD4+T细胞、RT-PCR法检测CD4+T细胞中Ets-1 mRNA及miR326的相对表达量。结果SLE组CD4+T细胞中miR326水平高于对照组(P=0.003),活动组高于稳定组(P=0.000);SLE组Ets-1的表达水平低于对照组(P=0.002),活动组低于稳定组(P=0.001);SLE组Th17细胞比例较对照组增高(P=0.004),活动组高于稳定组,差异有统计学意义;SLE患者miR326表达量与Ets-1表达量呈负相关性(P〈0.001),与Th17细胞比例呈正相关性(P=0.001);Ets-1与Th17细胞比例呈负相关性(P=0.003);SLE患者miR326的表达量及Th17细胞比例与SLEDAI评分呈正相关性(P〈0.001),Ets-1与SLEDAI评分呈负相关性(P〈0.001);SLE患者miR326水平及Th17细胞比例在皮疹、发热、血液系统及肾脏受累的患者中增高,Ets-1的表达量则降低;miR326水平及Th17细胞比例与抗ds DNA、抗核小体呈正相关性,与C3、C4呈负相关性,Ets-1与抗ds DNA、抗核小体呈负相关性,与C3、C4呈正相关性。结论 SLE患者CD4+T细胞中miR326表达明显增高并与Th17细胞比例、疾病活动指标呈正相关性,与Ets-1呈负相关性,提示miR326可能通过抑制Ets-1基因表达,促进Th17细胞分化参与SLE疾病活动。
Objective To detect the expression of miR326 and Ets-1 in CD4+T cells and the ratio of Th17 cells of patients with systemic lupus erythematosus( SLE),to analyze their relationship with each other and with the clinical data,to explore the potential pathogenesis of SLE. Methods 40 patients according to 2009 American College of Rheumatology( ACR) SLE diagnosis standards were selected,and recorded their clinical data. 14 healthy adults without autoimmune disease were chosen as controls. Flow cytometry was used to inspect the ratio of Th17 cells( CD4+IL-17A+) /CD4+T cells. Magnetic activated cell sorting was used to enrich the CD4+T cells,and reverse transcription polymerase chain reaction( RT-PCR) was used to inspect the relative expression of miR326 and Ets-1mRNA in CD4+T cells. Results Compared to the healthy controls,the miR326 expression level in the CD4+T cells of SLE patients was increased obviously( P = 0. 003),and the expression level was higher in the active group than that in remitting group( P = 0. 000). The Ets-1 mRNA expression level in CD4+T cells of SLE group was reduced obviously than that in the controls( P = 0. 002),and the expression in the active group was lower than that in remitting group( P = 0. 001). Compared to the controls,the ratio of Th17 cells / CD4+T cells in SLE group was increased obviously( P = 0. 004),and significant difference was found between the active group and the remitting group. The miR326 expression had a negative correlation with the Ets-1 expression( P〈 0. 001) and a positive correlation with the ratio of Th17 cells( P = 0. 001). There was also a negative correlation between Ets-1 expression and the ratio of Th17 cells( P = 0. 003). The miR326 expression and the ratio of Th17 cells both had a positive correlation with the SLEDAI score,while the Ets-1 expression had a negative correlation with the SLEDAI score. In SLE patients who suffered from erythra,hematologic system or kidney impairment,the miR326 expr