帕金森病和正常人外周血单个核细胞的蛋白质组差异分析

Proteome Analysis of Mononuclear Cell in Peripheral Blood between Patients with Parkinson Disease and Healthy Subjects

目的:通过研究帕金森病和正常外周血单个核细胞(PBMC)的蛋白质组差异,初步探讨外周免疫系统与帕金森病的病理联系方法:用固相pH梯度双向凝胶电泳分离人帕金森病和正常单个核细胞总蛋白质,考马斯亮蓝染色,PDQuest 2-DE软件分析,对部分差异蛋白质点进行基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)测定其胶内酶解后的肽质指纹图谱,用Mascot查询系统查询SWISS-PROT数据库。结果:获得了分辨率和重复性均较好的双向电泳考染图谱,对其中的21个差异蛋白质点分别进行肽质指纹分析,经数据库查询,初步鉴定为一些与蛋白降解、抗氧化应激、信号转导、细胞骨架、细胞周期调控等有关的蛋白质。结论:建立了帕金森病PBMC的双向凝胶电泳图谱,提示帕金森病和正常的PBMC的蛋白质表达具有差异。

Objective： To investigate the pathologic relationship between peripheral immunologic system and Parkinson Disease by comparison of proteomes difference of PBMC between health adults and Parkinson Disease＇s patients. Methods： The proteome of PBMC between health adults and Parkinson Disease＇s patients were separated and identified by a series of methods including immobilized pH gradient two-dimensional polyacrylamide gel electrophoresis （2-DE）, Coomassie Brilliant Blue staining, PDQuest 2-DE software analysis, peptide mass fingerprint analysis based on matrix-assisted laser adsorption/ionization time of flying mass spectrometry （MALDI-TOF-MS） and SWISS-PROT database searching, Results： The 2-DE pattern was obtained with high resolution rate and reproducibility. After Coomassie Brilliant Blue staining, 21 spots were incised and analyzed by MALDI-TOF-MS. The typical peptide masses were searched in the SWISS-PROT database by Mascot software. All typical protons were primarily identified to be related to cytoskeleton, anti-oxidative stress, protein degradation, signal transduction and cell cycle, etc. Conclusion： The 2-DE patterns of PD were established with well-resolution rate and reproducibility and the protein expressive difference of PBMC were revealed between PD patients and health adults.