系统性红斑狼疮外周血淋巴细胞5-羟色胺1A受体基因启动子区甲基化与mRNA表达的研究  

The study of DNA methylation status of HTR1A in the peripheral blood lymphocytes and the level of HTR1A mRNA in systemic lupus erythematosus patients

作　　者：徐健[1] 程宇琪[2] 赖爱云[1] 崔若玫[1] 吕昭萍[1] 许秀峰[2] 李路琼[1] 李姝[1] 白茹[1] 

机构地区：[1]昆明医科大学第一附属医院风湿免疫科,650032 [2]昆明医科大学第一附属医院精神科,650032

出　　处：《中华风湿病学杂志》2015年第9期586-591,共6页Chinese Journal of Rheumatology

基　　金：国家自然科学基金（81160379,81460256）;云南省医疗卫生单位内设研究机构项目基金（2014NS171.2014NS172）;云南省高层次卫生技术人才风湿免疫病学学科带头人基金（D-201218）;云南省创新团队项目（2014HC018）

摘　　要：目的 本研究通过比较117例SLE患者和117名匹配的健康志愿者外周血淋巴细胞（PBLC）中5-羟色胺1A（5-HT1A）受体基因（HTR1A）启动子区的甲基化状态及该基因mRNA的表达水平,探讨该基因启动子区的甲基化修饰与SLE的关系.方法 通过亚硫酸氢盐修饰克隆测序法检测目的基因的甲基化状态,用逆转录PCR检测目的基因mRNA的表达丰度.采用x2检验和Fisher确切概率法进行统计分析.结果 SLE患者PBLC中5-HT1A受体基因启动子区的甲基化程度,甲基化胞嘧啶总数（x^2=28.811,P＜0.01）和含甲基化胞嘧啶总人数（x^2=16.897,P＜0.01）均明显低于健康志愿者,呈现出一种低甲基化状态,尤其在-340位点的甲基化胞嘧啶的总数目SLE组明显少于健康者[1.7％（2/117）与9.4％（11/117）,x^2=6.597,P＜0.05];同时,SLE患者PBLC中5-HT1A受体基因的mRNA表达丰度明显高于健康者（43±16与0,t=12.82,P＜0.01）.结论 本研究结果支持5-HT1A受体基因启动子区的低甲基化及该基因的过度表达可能参与了SLE的病理机制,揭示了表观遗传调控可能在SLE的发病机制中起着重要的作用,提供了大脑通过5-羟色胺递质系统与免疫系统发生联系的证据.Objective To explore the DNA methylation status of the promoter region of HTR1A and the level of HTR1A mRNA in the peripheral blood lymphocytes of 117 systemic lupus erythematosus （SLE） patients and 117 matched healthy controls.Methods Bisulfite modification cloning sequencing was conducted to detect the DNA methylation status of the promoter region of HTR1A.reverse transcription polymerase chain reaction （RT-PCR） was used to test the level of HTR1A mRNA in the peripheral blood lymphocytes.Statistical analyses were performed using Chi-square test and Fisher＇s exact test.Results The results showed significant hypomethylation of the promoter region of HTR1A in SLE patients compared with the healthy controls （x^2=28.811,P〈0.01 for total mC and x^2=16.897,P〈0.01 for Person with mC）,espec.ially at site -340 [1.7% （2/117） vs 9.4% （11/117）,x^2=6.597,P〈0.05].The patients also showed a significantly higher HTR1A mRNA level than the controls （43±16 vs 0,t=12.82,P〈0.01）.Conclusion Our results support the hypothesis that the hypomethylation of the promoter region of HTR1A and overexpression of HTR1A might contribute to SLE.These results also reveal that epigenetic regulation via the serotonin system may contribute to SLE,and reveal the link between the brain and the immune system.

关 键 词：红斑狼疮 系统性 淋巴细胞 5-羟色胺1A受体基因 DNA甲基化 mRNA 信使 

分 类 号：R593.241[医药卫生—内科学]