中华风湿病学杂志
中華風濕病學雜誌
중화풍습병학잡지
CHINESE JOURNAL OF RHEUMATOLOGY
2009年
2期
98-102
,共5页
张丽%戴勇%胡成效%张艳亮
張麗%戴勇%鬍成效%張豔亮
장려%대용%호성효%장염량
红斑狼疮,系统性%组蛋白H3赖氨酸4%甲基化%染色质免疫共沉淀%芯片
紅斑狼瘡,繫統性%組蛋白H3賴氨痠4%甲基化%染色質免疫共沉澱%芯片
홍반랑창,계통성%조단백H3뢰안산4%갑기화%염색질면역공침정%심편
Lupus erythematosus,systemic%Histone H3 lysine 4%Methylation%Chromatin immunoprecipitation%Microarray
目的 研究系统性红斑狼疮(SLE)患者外周血单个核细胞(PBMCs)组蛋白H3赖氨酸4(H3K4)三甲基化(Me3)水平.方法 梯度密度离心法分离10例SLE活动期患者、7例SLE稳定期患者和8名健康者的PBMCs,采用染色质免疫共沉淀联合芯片技术(CHIP-chip)在全基因组范围内对SLE患者及健康者的PBMCs组蛋白H3K4me3进行高通量的筛选.染色质免疫共沉淀一实时定量聚合酶链反应(ChlP-qPCR)验证芯片结果.定量反转录一聚合酶链反应(qRT-PCR)检测H3K4me3显著差异基因的mRNA表达水平.结果 10例SLE活动期患者与健康对照相比较,鉴定出413个基因存在H3K4me3显著差异,其中137个基因显示H3K4me3程度增高,276个基因H3K4me3程度降低;7例SLE稳定期患者与健康对照相比较,发现393个基因存在H3K4me3表达差异,其中有112个基因H3K4me3程度增高,281个基因H3K4me3程度降低.ChIP-qPCR验证结果与CpG岛芯片的结果相一致.结论 SLE患者与健康者之间的PBMCs存在组蛋白H3K4me3显著改变.ChIP-chip技术有利于进一步揭示SLE分子机制,发现新的治疗靶点.
目的 研究繫統性紅斑狼瘡(SLE)患者外週血單箇覈細胞(PBMCs)組蛋白H3賴氨痠4(H3K4)三甲基化(Me3)水平.方法 梯度密度離心法分離10例SLE活動期患者、7例SLE穩定期患者和8名健康者的PBMCs,採用染色質免疫共沉澱聯閤芯片技術(CHIP-chip)在全基因組範圍內對SLE患者及健康者的PBMCs組蛋白H3K4me3進行高通量的篩選.染色質免疫共沉澱一實時定量聚閤酶鏈反應(ChlP-qPCR)驗證芯片結果.定量反轉錄一聚閤酶鏈反應(qRT-PCR)檢測H3K4me3顯著差異基因的mRNA錶達水平.結果 10例SLE活動期患者與健康對照相比較,鑒定齣413箇基因存在H3K4me3顯著差異,其中137箇基因顯示H3K4me3程度增高,276箇基因H3K4me3程度降低;7例SLE穩定期患者與健康對照相比較,髮現393箇基因存在H3K4me3錶達差異,其中有112箇基因H3K4me3程度增高,281箇基因H3K4me3程度降低.ChIP-qPCR驗證結果與CpG島芯片的結果相一緻.結論 SLE患者與健康者之間的PBMCs存在組蛋白H3K4me3顯著改變.ChIP-chip技術有利于進一步揭示SLE分子機製,髮現新的治療靶點.
목적 연구계통성홍반랑창(SLE)환자외주혈단개핵세포(PBMCs)조단백H3뢰안산4(H3K4)삼갑기화(Me3)수평.방법 제도밀도리심법분리10례SLE활동기환자、7례SLE은정기환자화8명건강자적PBMCs,채용염색질면역공침정연합심편기술(CHIP-chip)재전기인조범위내대SLE환자급건강자적PBMCs조단백H3K4me3진행고통량적사선.염색질면역공침정일실시정량취합매련반응(ChlP-qPCR)험증심편결과.정량반전록일취합매련반응(qRT-PCR)검측H3K4me3현저차이기인적mRNA표체수평.결과 10례SLE활동기환자여건강대조상비교,감정출413개기인존재H3K4me3현저차이,기중137개기인현시H3K4me3정도증고,276개기인H3K4me3정도강저;7례SLE은정기환자여건강대조상비교,발현393개기인존재H3K4me3표체차이,기중유112개기인H3K4me3정도증고,281개기인H3K4me3정도강저.ChIP-qPCR험증결과여CpG도심편적결과상일치.결론 SLE환자여건강자지간적PBMCs존재조단백H3K4me3현저개변.ChIP-chip기술유리우진일보게시SLE분자궤제,발현신적치료파점.
Objective To study histone H3 lysine 4 trimethylation (H3K4me3) in peripheral blood mononuclear cells (PBMCs) of systemic lupus erythematosus(SLE) patients.Methods PBMCs were isolated by density gradient centrifugation from 10 active SLE patients,7 inactive SLE patients and 8 healthy volunteers.Chromatin immunopreeipitation linked to mieroarrays (ChIP-chip) was used to profile the variations in H3K4me3 in CpG island regions in PBMCs of SLE patients and controls.ChIP-qPCR was used to validate the mieroarray results.To confirm correlations between H3K4me3 and gene expression,expression analysis by qRT-PCR was performed on three randomly selected H3K4me3 candidates.Results 413 (137 increased and 276 decreased H3K4me3) and 393 genes (112 increased and 281 decreased H3K4me3) displayed significant differences in H3K4me3 between active and inactive SLE when compared with healthy SUhjeets.The results of ChiP-qPCR were consistent with microarray.ConclusiOn There are significant differences in H3K4me3 profiling between SLE and healthy subjects.These novel candidate genes may be potential biomarkers for future therapeutic targets.The ChIP-chip technology can help further reveal SLE molecular mechanisms and discover new therapeutic targets.
