中华风湿病学杂志
中華風濕病學雜誌
중화풍습병학잡지
CHINESE JOURNAL OF RHEUMATOLOGY
2013年
4期
264-267
,共4页
郝轶群%刘秀梅%闫欣%杨洁%傅自力%罗东萍%王凯
郝軼群%劉秀梅%閆訢%楊潔%傅自力%囉東萍%王凱
학질군%류수매%염흔%양길%부자력%라동평%왕개
关节炎,类风湿%T淋巴细胞,辅助诱导%单个核细胞%Foxp3
關節炎,類風濕%T淋巴細胞,輔助誘導%單箇覈細胞%Foxp3
관절염,류풍습%T림파세포,보조유도%단개핵세포%Foxp3
Arthritis,rheumatoid%T-lymphocytes,helper-inducer%Peripheral blood mononuclear cells%Foxp3
目的 通过检测类风湿关节炎(RA)患者外周血单个核细胞(PBMCs)中CD4+CD25+调节性T细胞Foxp3的表达水平,以及Foxp3基因启动子区甲基化情况,探讨其在RA发病机制中的作用.方法 选取25例RA患者和10名健康对照,用密度梯度离心法提取外周血单个核细胞(PBMCs)；采用流式细胞仪检测CD4+CD25+调节性T细胞Foxp3的表达水平；实时荧光定量聚合酶链反应(PCR)检测Foxp3 mRNA在PBMCs中的表达情况；应用重亚硫酸盐处理基因测序法测定PBMCs中Foxp3基因启动子序列甲基化水平的差异.采用单因素方差分析和Fisher确切概率法进行统计分析.结果 RA活动组CD4+CD25+调节性T细胞Foxp3 mRNA的表达水平(2.31±0.25)显著低于RA非活动组(3.68±0.26)和健康对照组(5.67±0.34),差异有统计学意义(P＜0.05)；RA非活动组Foxp3 mRNA的表达水平低于健康对照组(P＜0.05).而RA患者PBMCs中Foxp3启动子区-67、-74位点甲基化水平(46％)明显高于健康对照组(6％),差异有统计学意义(P＜0.05).结论 CD4+CD25+调节性T细胞数量的减少可能参与RA的发病,而在此过程中Foxp3基因启动子区甲基化水平起到关键作用.
目的 通過檢測類風濕關節炎(RA)患者外週血單箇覈細胞(PBMCs)中CD4+CD25+調節性T細胞Foxp3的錶達水平,以及Foxp3基因啟動子區甲基化情況,探討其在RA髮病機製中的作用.方法 選取25例RA患者和10名健康對照,用密度梯度離心法提取外週血單箇覈細胞(PBMCs)；採用流式細胞儀檢測CD4+CD25+調節性T細胞Foxp3的錶達水平；實時熒光定量聚閤酶鏈反應(PCR)檢測Foxp3 mRNA在PBMCs中的錶達情況；應用重亞硫痠鹽處理基因測序法測定PBMCs中Foxp3基因啟動子序列甲基化水平的差異.採用單因素方差分析和Fisher確切概率法進行統計分析.結果 RA活動組CD4+CD25+調節性T細胞Foxp3 mRNA的錶達水平(2.31±0.25)顯著低于RA非活動組(3.68±0.26)和健康對照組(5.67±0.34),差異有統計學意義(P＜0.05)；RA非活動組Foxp3 mRNA的錶達水平低于健康對照組(P＜0.05).而RA患者PBMCs中Foxp3啟動子區-67、-74位點甲基化水平(46％)明顯高于健康對照組(6％),差異有統計學意義(P＜0.05).結論 CD4+CD25+調節性T細胞數量的減少可能參與RA的髮病,而在此過程中Foxp3基因啟動子區甲基化水平起到關鍵作用.
목적 통과검측류풍습관절염(RA)환자외주혈단개핵세포(PBMCs)중CD4+CD25+조절성T세포Foxp3적표체수평,이급Foxp3기인계동자구갑기화정황,탐토기재RA발병궤제중적작용.방법 선취25례RA환자화10명건강대조,용밀도제도리심법제취외주혈단개핵세포(PBMCs)；채용류식세포의검측CD4+CD25+조절성T세포Foxp3적표체수평；실시형광정량취합매련반응(PCR)검측Foxp3 mRNA재PBMCs중적표체정황；응용중아류산염처리기인측서법측정PBMCs중Foxp3기인계동자서렬갑기화수평적차이.채용단인소방차분석화Fisher학절개솔법진행통계분석.결과 RA활동조CD4+CD25+조절성T세포Foxp3 mRNA적표체수평(2.31±0.25)현저저우RA비활동조(3.68±0.26)화건강대조조(5.67±0.34),차이유통계학의의(P＜0.05)；RA비활동조Foxp3 mRNA적표체수평저우건강대조조(P＜0.05).이RA환자PBMCs중Foxp3계동자구-67、-74위점갑기화수평(46％)명현고우건강대조조(6％),차이유통계학의의(P＜0.05).결론 CD4+CD25+조절성T세포수량적감소가능삼여RA적발병,이재차과정중Foxp3기인계동자구갑기화수평기도관건작용.
Objective By detecting the expression levels of Foxp3 in CD4+CD25+ regulatory T cells of peripheral blood mononuclear cells (PBMCs) in rheumatoid arthritis (RA),and the Foxp3 gene promoter region methylation to explore its role in the pathogenesis of RA.Methods Twenty-five RA patients and 10 healthy controls were selected,and the PBMCs were extracted by density gradient centrifugation.Foxp3 expression levels of CD4+CD25+ regulatory T cells were detected by flow cytometry.The real-time fluorescence quantitative PCR assay was used to detect the Foxp3 mRNA expression in PBMCs; and bisulfate processing gene sequencing was used to determinethe differences in Foxp3 gene promoter sequence methylation level of PBMCs.The comparison between groups was analyzed using one-way ANOVA; two sets of qualitative data were compared using Fisher's exact test.Results The expression levels of Foxp3 mRNA in the CD4+CD25+regulatory T cells of active RA patients (2.31±0.25) was significantly lower than inactive RA group (3.68±0.26) and healthy controls (5.67±0.34),the difference was statistically significant (P＜0.05).The Foxp3 mRNA expression level in inactive RA group was lower than that of the healthy controls (P＜0.05).Foxp3 promoter region-67,-74 sites of methylation level in PBMCs of RA patients (46％) was significantly higher than that of the healthy controls (6％).Conclusion Reduction in the number of CD4+CD25+ regulatory T cells may be involved in the pathogenesis of RA and Foxp3 gene promoter methylation levels plays a key role in this process.
