趋化因子CXCL14在SLE患者外周血单个核细胞中的表达及启动子甲基化分析

doi:10.12047/j.cjap.5415.2017.049

中国应用生理学杂志 ›› 2017, Vol. 33 ›› Issue (3): 197-201.doi: 10.12047/j.cjap.5415.2017.049

趋化因子CXCL14在SLE患者外周血单个核细胞中的表达及启动子甲基化分析

王金丹¹, 黄茜茜², 叶璐璐³, 范超凡², 郭刚强³, 徐玲娟⁴, 薛向阳³, 盛秀胜⁵

1. 温州医科大学检验医学院、生命科学院, 浙江温州 325035;
2. 温州医科大学第二临床医学院, 浙江温州 325035;
3. 温州医科大学微生物学和免疫学教研室, 分子病毒与疫苗研究所, 热带医学研究所, 浙江温州 325035;
4. 金华市中医医院检验科, 浙江金华 321017;
5. 金华职业技术学院医学院, 浙江金华 321017

收稿日期:2016-01-15 修回日期:2016-11-29 出版日期:2017-05-28 发布日期:2018-06-20
通讯作者: 盛秀胜,Tel:0579-82232886,E-mail:shengsheng117@sina.com E-mail:shengsheng117@sina.com
基金资助:
浙江省科技厅科研基金资助项目（2012C33126）；浙江省大学生新苗计划（2014R413058，2015R413027，2015R413069）；金华市科技局科研基金资助项目（2013-3-014）

Expression and its promoter methylation of chemokine CXC ligand 14 in peripheral blood mononuclear cells from patients with lupus

WANG Jin-dan¹, HUANG Qian-qian², YE Lu-lu³, FAN Chao-fan², GUO Gang-qiang³, XU Ling-juan⁴, XUE Xiang-yang³, SHENG Xiu-sheng⁵

1. School of Laboratory Medicine and Life Science, Wenzhou Medical University, Wenzhou 325035;
2. The Second Clinical College, Wenzhou Medical University, Wenzhou 325035;
3. Department of Microbiology and Immunology, Institute of Molecular Virology and Immunology, Institute of Tropical Medicine, Wenzhou Medical University, Wenzhou 325035;
4. Department of Laboratory, Jinhua Hospital of Traditional Chinese Medicine, Jinhua 321000, China;
5. Medical School, Jinhua Polytechnic College, Jinhua 321000, China

Received:2016-01-15 Revised:2016-11-29 Online:2017-05-28 Published:2018-06-20
Supported by:
浙江省科技厅科研基金资助项目（2012C33126）；浙江省大学生新苗计划（2014R413058，2015R413027，2015R413069）；金华市科技局科研基金资助项目（2013-3-014）

摘要/Abstract

摘要： 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CX-CL14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CpG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CpG岛过度甲基化是SLE患者CXCL14低表达的重要机制。

关键词: 系统性红斑狼疮, 趋化因子, CXCL14, 甲基化

Abstract: Objective: To analyze the expression and its promoter methylation of chemokine CXC ligand 14 (CXCL14) in peripheral blood mononuclear cells (PBMCs) from patients with systemic lupus erythematosus (SLE). Methods: The RNAs of PBMCs from 28 SLE patients and 20 healthy controls were isolated and reversely transcribed into cDNAs. Using GAPDH as the internal reference, the levels of CXCL14 ex-pression were detected by real-time polymerase chain reaction (PCR). The correlation between CXCL14 expression and the clinic pathological fe atures of SLE were further analyzed. DNA methylation was analyzed by bisulfite sequencing PCR (BSP). Results: Our data indicated that the level of CXCL14 in the PBMC of SLE patients was statistically lower than that in healthy controls (P < 0.05). Further analysis showed that CXCL14 expression was negatively correlated with anti-Sj gren syndrome B antibody(anti-SSB antibody, P < 0.01) and albuminuria(P < 0.05). However, CXCL14 expression was not significantly correlated with the indexes of SLE activity, renal damage, the level of anti-ds-DNA antibodies, complement C3 and C-reactive protein. In addition, we further demonstrated that the CXCL14 promoter hypermethylation expres-sion was significant higher than healthy controls. Conclusion: Down-regulated of CXCL14 expression in PBMC maybe involved in the occur-rence or development of SLE disease. The loss of CXCL14 expression was regulated by promoter hypermethylation.

Key words: systemic lupus erythematosus (SLE), chemokine, CXCL14, methylation

王金丹, 黄茜茜, 叶璐璐, 范超凡, 郭刚强, 徐玲娟, 薛向阳, 盛秀胜. 趋化因子CXCL14在SLE患者外周血单个核细胞中的表达及启动子甲基化分析[J]. 中国应用生理学杂志, 2017, 33(3): 197-201.

WANG Jin-dan, HUANG Qian-qian, YE Lu-lu, FAN Chao-fan, GUO Gang-qiang, XU Ling-juan, XUE Xiang-yang, SHENG Xiu-sheng. Expression and its promoter methylation of chemokine CXC ligand 14 in peripheral blood mononuclear cells from patients with lupus[J]. CJAP, 2017, 33(3): 197-201.

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趋化因子CXCL14在SLE患者外周血单个核细胞中的表达及启动子甲基化分析

Expression and its promoter methylation of chemokine CXC ligand 14 in peripheral blood mononuclear cells from patients with lupus

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