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,PDQuest2-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.