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目的:对比分析系统性红斑狼疮肾炎(LN)患者和健康人双重滤过血浆置换(DFPP)产物中的差异蛋白质,筛选与LN发生相关的血清蛋白质。方法:利用DFPP的方法分别对4例LN患者和4例健康人血浆进行过滤,将滤下的大分子蛋白混合物行双向电泳分离并银染显影,经图像分析筛选差异表达的蛋白点。应用QSTAR Pulsar I型串连飞行质谱鉴定差异表达的蛋白点,通过生物信息学比对,对其中有意义分子行血清浓度检测验证。结果:2组间筛选出差异蛋白点共317个,其中表达量差异达3倍以上的蛋白点69个,经串连质谱分析鉴定出包括人冷凝集素IgM抗体、免疫球蛋白λ轻链、谷胱甘肽-S-转移酶、结合珠蛋白前原蛋白、甲状腺素转运蛋白、载脂蛋白E、皮质类固醇结合球蛋白、玻连蛋白、角蛋白和转铁蛋白等十多个在SLE活动性肾炎血清中表达改变的蛋白分子。ELISA及RIA检测结果与双向电泳相符。结论:LN患者血清与健康人血清的蛋白表达谱有一定差异,这些差异分子可能成为LN诊断及预后判断的重要标志物。
OBJECTIVE: To compare and analyze the differential proteins in the double-filtration plasmapheresis (DFPP) of patients with systemic lupus erythematosus nephritis (LN) and healthy people and screen the serum proteins related to the occurrence of LN. Methods: The plasma of 4 LN patients and 4 healthy individuals were filtered by DFPP method. The separated macromolecule protein mixture was separated by two - dimensional electrophoresis and developed by silver staining. The differentially expressed protein spots were screened by image analysis. The differentially expressed protein spots were identified by QSTAR Pulsar type I tandem mass spectrometry. Bioinformatics analysis was used to validate the serum concentrations of some of them. Results: A total of 317 differentially expressed protein spots were screened out from the two groups, of which 69 were differentially expressed more than 3 times. The results of tandem mass spectrometry (LC-MS / MS) showed that IgM antibody including human cold agglutinin, immunoglobulin lambda light chain, Glutathione-S-transferase, haptoglobin, thyroxine transporter, apolipoprotein E, corticosteroid-binding globulin, vitronectin, keratin and transferrin in more than 10 in the SLE activity Nephritis serum expression of altered protein molecules. ELISA and RIA results consistent with two-dimensional electrophoresis. CONCLUSION: The protein expression profiles of LN patients and healthy individuals are different. These differentially expressed molecules may be important markers in the diagnosis and prognosis of LN.