定量蛋白质组学技术构建系统性红斑狼疮疾病的蛋白质组差异表达图谱

摘要

AIM : To identify and quantify the total proteins in peripheral hlood mononuclear cells ( PBMC ) of systemic lupus erythematosus ( SLE ) patients with quantitative proteomic technique, and to establish a differential expression profile of proteome for SLE.METHODS: Four - plex isobaric tags for relative and absolute quantification coupled with multiple chromatographic fractionation and tandem mass spectrometry were used to analyze the total proteins in PBMC from healthy controls and the patients of stable SLE, active SLE and theumatoid arthritis.The proteins were identified by datahase searching with peptide mass fingerprinting.The differential expression of the proteins was compared.RESULTS :More than 400 proteins were identified.Compared with healthy controls , 44 proteins were discovered to be significantly expressed hy more than 2 folds in stahle SLE and active SLE, among which 9 proteins were up - regulated and 35 proteins were down - regulated.Compared with theumatoid arthritis group, 52 proteins displayed 2 or more folds of changes in stable SLE and active SLE, including 19 up - regulated proteins and 33 down - regulated ones.The up - and down - regulated proteins between active SLE and stable SLE were 17 and 13 , respectively.CONCLUSION : Quantitative proteomic technique is efficiently applicable for protein identification and relative quantitation in human peripheral blood mononuclear cells.Determination of the differentially expressed proteomic profile of SLE is helpful for better understanding the pathogenesis of SLE and developing new strategies for diagnosis and treatment of SLE.%目的:应用定量蛋白质组学技术对系统性红斑狼疮(SLE)病人外周血单个核细胞中的"全组"蛋白进行鉴定和定量分析,获得SLE的蛋白质组差异表达图谱.方法:利用基于四重相对和绝对定量的等量异位标签结合多维液相色谱-串联质谱分析SLE稳定期和活动期病人以及类风湿关节炎病人和健康人的外周血单个核细胞总蛋白,用肽质量指纹谱经数据库检索鉴定蛋白质,并比较这些蛋白的表达差异.结果:共鉴定了400多个蛋白质.其中,与健康对照组相比,SLE稳定组和SLE活动组共发现2倍以上表达差异的蛋白质44个,上调的9个,下调的35个;与类风湿关节炎疾病组相比,SLE稳定组和SLE活动组2倍以上表达差异的蛋白共有52个,上调的19个,下调的33个;SLE活动组与SLE稳定组相比,表达上调和下调的蛋白分别为17个和13个.结论:定量蛋白质组学技术可有效地用于人外周血单个核细胞蛋白鉴定和相对定量;采用该技术获得了SLE病人外周血单个核细胞的蛋白质组差异表达图谱;深入研究这些蛋白的分子机制将有助于进一步阐明SLE的发病机制,为SLE的诊断和治疗提供新途径.