系统性红斑狼疮患者血清蛋白质组学研究

Identification of Biomarkers in Serum of Systemic Lupus Erythematosus by Proteomic Method

目的运用蛋白质组学的方法,分析正常人及SLE患者血清蛋白质的差异表达,寻找与SLE疾病发病机制相关的蛋白质。方法分别收集健康人及SLE患者血清各9例,同组血清等量混合,用试剂盒除去血清中的白蛋白和免疫球蛋白,再经除盐浓缩后,将血清样品采用固相pH梯度(IPG)双向凝胶电泳(2-DE)分离正常人及SLE患者血清的总蛋白质。凝胶经考马斯亮蓝染色显色后,利用Analysis2d软件对获得的蛋白图谱进行分析,寻找差异表达的蛋白质,利用基质辅助激光解析电离飞行时间质谱(MALDI-TOF-MS)进行鉴定,分析差异蛋白点。结果对照组凝胶共检出蛋白点648个,患者组检出639个。对照组凝胶蛋白点匹配率84.5%,患者组凝胶蛋白点匹配率82.5%。通过比较分析,差异表达蛋白质点数为92个,有52个蛋白点在SLE患者组表达上调,40个表达下调,有14个点的表达水平在组间差异有统计学意义,质谱鉴定共鉴定5个蛋白质。通过文献研究显示,我们鉴定的部分蛋白在SLE的发病机制中起潜在的作用。结论在SLE患者血清中存在着差异血清蛋白质,这些蛋白质可能是SLE发病的内在因素,并且在SLE的疾病发展过程中发挥重要作用,可能作为新的血清标志物和潜在的自身抗原。

Objective To discover the biomarkers of serum of systemic lupus erythematous by proteomic methods.Methods Peripheral blood was obtained from 9 SLE patients and 9 healthy controls and then mononuclear cells were isolated and the total protein was extracted by one-step method.Two-dimensional gel electrophoresis was performed and then stained with silver.Protein maps were analyzed and differentially expressed protein spots were detected using Image Master 2D Platinum5.0 software.Results Match rates of 84.5％ and 82.5％ was obtained from gels from controls and patients respectively.648 spots were detected from control gels and 639from patient gels.8 protein pots were up regulated and 6 were down-regulated in SLE patients.5 proteins were identified by MS analysis,some of which had previously been shown to play a potential role in the pathogenesis of SLE.Conclusion There are significant changes in the protein expression of peripheral blood mononuclear cells in systemic lupus erythematosus patients.This study could be used as a preliminary work for better understanding of the pathogenesis and immune regulation pathways of SLE from integrated lymphocyte protein profile perspective.