蛋白质组学技术筛选破裂颅内动脉瘤的血清标志物

Identification of serum biomarkers from ruptured intracranial aneurysm patients in Chinese using proteomics pattern

目的探讨用双向凝胶电泳结合基质辅助激光解析离子化飞行时间质谱(MALDI-TOF MS)分析技术从血清中筛选颅内动脉瘤标志蛋白的可行性。方法分别采集6例破裂颅内动脉瘤患者和健康成人的血清,进行双向凝胶电泳分离血清总蛋白,考马斯亮蓝染色,全自动斑点处理工作站处理差异蛋白质斑点,MALDI-TOF MS分析获取肽质量指纹图谱,对鉴定的差异蛋白质进行分析。结果两组表达差异2倍以上的蛋白质斑点有81个,质谱鉴定出5个差异蛋白质,在颅内动脉瘤血清中均呈上调表达,即α2-巨球蛋白前体、补体3前体、玻璃黏连蛋白前体、缓激肽和载脂蛋白E3。结论蛋白质组学技术平台为大规模水平筛选破裂颅内动脉瘤血清标志物提供了新方法。

Objective To investigate the protein composition of the serum from patients with intracranial aneurysm in Chinese using proteomics approach based on two-dimensional （2-D） gel electrophoresis and MALDI-TOF mass spectrometric technique. Methods We analyzed serum differential protein expression from six ruptured intracranial aneurysm patients and six healthy donors. The serum samples after removal of Albumin/IgG were subjected 2-D gel electrophoresis with first dimension IEF and second dimension SDS-PAGE. Gels were stained using Coomassie Brilliant Blue and digitized in Imagescanner II image scanner. Images were analyzed densitometrically by ImageMaster 2-D Elite Software 4.9. The SDS-PAGE gels were placed into the Ettan Spot Handling Workstation and the pick list was input into the computer. The differential protein spots were automatically excised from the gels, washed, dried and digested by trypsin. The obtained peptides were put into EttanTM MALDI-TOF Mass Spectrometry and then the peptide mass list was submitted to NCBInr database using MALDI EVALUATION V2.0 software to search and identify possible target proteins. Results High-quality 2-D gel pattern were obtained in this study. We found 81 differential protein spots between intracranial aneurysm patients and healthy donors, only 5 spots had statistical difference. Of the mapped protein spots, we could identify 5 spots by mass spectrometry and database search, representing 5 individual proteins which included alpha 2 macroglobulin precursor, complement component 3 precursor, vitronectin precursor, bradykinin, Apoliprotein E3. Conclusions Using proteomics technique based on 2-D gel electrophoresis and MALDI-TOF mass spectrometry may be a useful approach to identify serum biomarkers from intracranial aneurysm patients in the future.