CAJ | 학술논문

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(x2=28.811,P＜0.01)和含甲基化胞嘧啶总人数(x2=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),x2=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.
목적 본연구통과비교117례SLE환자화117명필배적건강지원자외주혈림파세포(PBLC)중5-간색알1A(5-HT1A)수체기인(HTR1A)계동자구적갑기화상태급해기인mRNA적표체수평,탐토해기인계동자구적갑기화수식여SLE적관계.방법 통과아류산경염수식극륭측서법검측목적기인적갑기화상태,용역전록PCR검측목적기인mRNA적표체봉도.채용x2검험화Fisher학절개솔법진행통계분석.결과 SLE환자PBLC중5-HT1A수체기인계동자구적갑기화정도,갑기화포밀정총수(x2=28.811,P＜0.01)화함갑기화포밀정총인수(x2=16.897,P＜0.01)균명현저우건강지원자,정현출일충저갑기화상태,우기재-340위점적갑기화포밀정적총수목SLE조명현소우건강자[1.7％(2/117)여9.4％(11/117),x2=6.597,P＜0.05];동시,SLE환자PBLC중5-HT1A수체기인적mRNA표체봉도명현고우건강자(43±16여0,t=12.82,P＜0.01).결론 본연구결과지지5-HT1A수체기인계동자구적저갑기화급해기인적과도표체가능삼여료SLE적병리궤제,게시료표관유전조공가능재SLE적발병궤제중기착중요적작용,제공료대뇌통과5-간색알체질계통여면역계통발생련계적증거.
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 (x2=28.811,P＜0.01 for total mC and x2=16.897,P＜0.01 for Person with mC),espec.ially at site -340 [1.7％ (2/117) vs 9.4％ (11/117),x2=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.