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目的:分析趋化因子CXCL14在系统性红斑狼疮(SLE)患者外周血单个核细胞(PBMC)中的表达及其启动子甲基化特征。方法:收集28例SLE患者和20名健康人外周血单个核细胞(PBMC)标本,抽提细胞中RNA,逆转录后,以GAPDH为内参,通过定量PCR检测PBMC中CXCL14的表达,统计分析CXCL14表达水平与SLE各种临床资料的相关性,探讨PBMC中CXCL14表达与SLE的关系,通过重亚硫酸盐修饰的全基因组DNA用以BSP测序来明确不同标本中CXCL14启动子中甲基化位点的甲基化率。结果:CXCL14在SLE患者和正常人PBMC中的表达量存在显著差异(P<0.05)。与健康人PBMC中CXCL14的含量相比较,CXCL14在SLE患者PBMC中表达量显著降低。与进一步CXCL14表达水平与SLE各种临床资料的相关性分析显示,CXCL14表达水平与SSB抗体(干燥综合征B抗体)、蛋白尿及血小板计数相关。与SSB抗体阴性SLE患者比较,SSB抗体阳性患者CXCL14表达水平更低(P<0.05);与蛋白尿阴性SLE患者相比,蛋白尿阳性患者CXCL14表达水平更低(P<0.05);而血小板升高患者的CXCL14表达水平更高(P<0.05)。CXCL14表达水平与SLE活动、肾损害指标、抗ds-DNA、C反应蛋白(CRP)、补体C3水平等指标未见显著相关性。CXCL14启动子区甲基化分析显示,SLE患者Cp G岛甲基化明显高于正常对照。结论:PBMC中低表达的CXCL14与SLE发生发展相关,其启动子区Cp G岛过度甲基化是SLE患者CXCL14低表达的重要机制。
OBJECTIVE: To analyze the expression and promoter methylation of chemokine CXCL14 in peripheral blood mononuclear cells (PBMC) from patients with systemic lupus erythematosus (SLE). Methods: Totally 28 peripheral blood mononuclear cells (PBMCs) from SLE patients and 20 healthy controls were collected for RNA extraction and RT-PCR. GAPDH was used as an internal control to detect the expression of CXCL14 in PBMCs by quantitative PCR. The expression of CXCL14 CXCL14 expression in PBMC and the relationship between SLE, whole-genome DNA modified by bisulfite by BSP sequencing to clear different samples of CXCL14 promoter methylation sites The methylation rate. Results: CXCL14 expression in SLE patients and normal PBMC were significantly different (P <0.05). Compared with the content of CXCL14 in PBMC of healthy people, the expression level of CXCL14 in PBMC of SLE patients was significantly decreased. Correlation analysis between the expression of CXCL14 and various clinical data of SLE showed that the expression level of CXCL14 correlated with SSB antibody (Sjogren’s syndrome antibody), proteinuria and platelet count. Compared with patients with SSB negative SLE, CXCL14 expression was lower in SSB positive patients (P <0.05); CXCL14 expression was lower in proteinuric positive patients than those with negative proteinuria (P <0.05) Patients with high CXCL14 expression levels were higher (P <0.05). CXCL14 expression levels and SLE activity, renal damage indicators, anti-ds-DNA, C-reactive protein (CRP), complement C3 levels and other indicators no significant correlation. CXCL14 promoter region methylation analysis showed that SLE patients with Cp G island methylation was significantly higher than the normal control. CONCLUSIONS: The low expression of CXCL14 in PBMC correlates with the occurrence and development of SLE. The hypermethylation of CpG island in promoter region is an important mechanism of low expression of CXCL14 in SLE patients.