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目的探讨系统性红斑狼疮患者(SLE)T淋巴细胞TNFSF7基因启动子区域DNA甲基化状态及其在发病机制中的作用。方法分离15例活动期SLE患者、15例非活动期SLE患者和15例正常人对照外周血CD4+与CD8+T细胞,并分别提取DNA。采用亚硫酸氢钠测序法对CD4+与CD8+T细胞TN-FSF7基因启动子区域DNA甲基化水平进行检测。结果在TNFSF7基因启动子区域,活动期、非活动期SLE患者组的CD4+T淋巴细胞的TNFSF7基因启动子序列-600～-300bp区域平均甲基化水平(0.32±0.05,0.36±0.05)明显低于正常人对照组(0.62±0.05,P=0.000),且活动期平均甲基化水平明显低于非活动期SLE患者组(P=0.000)。活动期、非活动期SLE患者组的CD8+T淋巴细胞的TN-FSF7基因启动子序列平均甲基化水平与正常人比较差异无统计学意义(P>0.05)。结论活动期、非活动期SLE患者的CD4+T淋巴细胞的TNFSF7基因启动子区域处于低甲基化状态,且启动子甲基化水平与SLE的活跃性相关。
Objective To investigate the DNA methylation status of TNFSF7 gene promoter region in systemic lupus erythematosus (SLE) T lymphocyte and its role in pathogenesis. Methods Peripheral blood CD4 + and CD8 + T cells were isolated from 15 active SLE patients, 15 inactive SLE patients and 15 normal controls, and DNA was extracted separately. The DNA methylation level of TN-FSF7 promoter region in CD4 + and CD8 + T cells was detected by sodium bisulfite sequencing. Results In the TNFSF7 gene promoter region, the mean methylation level of TNFSF7 gene promoter region (-600-300 bp) in CD4 + T lymphocytes of active and inactive SLE patients (0.32 ± 0.05, 0.36 ± 0.05) was significantly (0.62 ± 0.05, P = 0.000), and the average methylation level in active stage was significantly lower than that in inactive SLE patients (P = 0.000). The average methylation level of TN-FSF7 gene promoter in CD8 + T lymphocytes of inactive and inactive SLE patients was not significantly different from that of normal controls (P> 0.05). Conclusions In active and inactive SLE patients, the promoter region of TNFSF7 gene in CD4 + T lymphocytes is hypomethylated, and the promoter methylation level is correlated with the activity of SLE.