年龄相关性白内障和正常晶状体蛋白质组的差异分析

Difference analysis of proteome between age-related cataract and normal lens

目的运用双向电泳和基质辅助的激光解吸电离飞行时间质谱(matrix-assisted laser desorption/ionization time-of-flight mass spectrometry,MALDI-TOF-MS)的方法,鉴定年龄相关性白内障和正常晶状体之间差异表达的蛋白质。方法用固相pH梯度双向凝胶电泳技术分离16例年龄相关性白内障和正常晶状体之间的总蛋白质,建立二维电泳图谱,再利用MALDI-TOF-MS分析及数据库搜索相结合对差异表达的蛋白质点进行鉴定。结果双向电泳胶图显示年龄相关性白内障和正常晶状体共有84个差异点,其中52个上调表达,32个下调表达。有和无的差异表达点10个,相差3倍以上差异表达点10个。利用MALDI-TOF-MS对差异表达的15个蛋白质点进行肽指纹图谱分析,经数据库检索后鉴定出5个蛋白质。结论年龄相关性白内障和正常晶状体之间蛋白表达存在差异,蛋白质组的分析结果为年龄相关性白内障的基础研究提供依据和新实验方法。

Objective To identify the different expressed proteins between the normal lens and age-related cataract by two-dimensional gel electrophoresis （2-DGE） and matrix-assisted laser desorption /ionization time-of-flight mass spectrometry（MALDI-TOF-MS）. Methods The proteins in normal lens （8 cases） and age-related cataract （8 cases） was analyzed by 2-DGE. Some different protein spots were identified by MALDI-TOF-MS. Results Eighty-four different spots were detected in 2-DGE, upgrading expressed in 52 spots and downgrading expressed in 32 spots. There were 10 spots in age-related cataract reproducibly altered more than 3-fold compared to normal controls,and 10 spots were only appeared in age-related cataract or normal lens. Fifteen protein spots were identified by MALDI-TOF-MS and five differential proteins were positively identiffed by peptide mass Fingerprinting. Conclusion The results provides base for future studies on age-related cataract-associated proteins. The proteomic approach is essential to understand pathogenesis and development of age-related cataract,and it is expected to obtain some clues for further study on the mechanism and new drug exploitation of cataract.