iTRAQ联合质谱技术研究孕期惊恐刺激下仔鼠脑发育的差异表达蛋白  

Study on differentially expressed proteins about brain development of newborn mice stimulated by fear during pregnancy by iTRAQ combined with mass spectrometry

作　　者：杨丽萍[1] 陈蕾[1] 陈四清[1] 李新民[1] 高爱社[1] 詹向红[1] 侯俊林[1] 

机构地区：[1]河南中医学院中西医结合基础学科,河南郑州450046

出　　处：《中国妇幼保健》2014年第21期3505-3508,共4页Maternal and Child Health Care of China

基　　金：河南省科技创新人才项目(114100510011);河南省教育厅自然科学项目(2010A360027);国家自然科学基金项目(81072734);国家自然科学基金项目(81373790)

摘　　要：目的:应用蛋白组学筛选孕期惊恐刺激下的模型组仔鼠和对照组仔鼠海马组织中的差异表达蛋白质,分析其与脑发育的关系。方法:采集模型组和对照组仔鼠海马组织提取蛋白质,进行蛋白变性、还原及酶解,最后采用核素标记相对和绝对定量技术(isobaric tags for relative and absolute quantitation,iTRAQ)试剂标记,液相色谱串联质谱技术(liquid chromatography-tandem mass spectrometry,LC-MS/MS)筛选鉴定差异表达蛋白,对差异表达蛋白进行生物学分析。结果:质谱共鉴定出惊恐刺激下仔鼠海马组织蛋白质215个,符合条件的差异蛋白质158个,58个蛋白质表达上调,100个蛋白质表达下调。158个差异表达蛋白质共参与150种生物过程、48种细胞组分和52种分子功能。蛋白质交互网络图显示Ctnnb1、Vcl和14-3-3蛋白质家族(Ywhag、Ywhae、Ywhah、Ywhaq、Ywhab)处于网络图主要交叉点。结论:处于蛋白质交互网络主要交叉点位置的蛋白质可能在孕期惊恐刺激下仔鼠脑发育过程中发挥重要作用。Objective： To screen differentially expressed proteins in hippocampal tissues of newborn mice stimulated by fear during pregnancy in model group and control group by proteomic method, analyze its relationship with brain development. Methods： Proteins were abstracted from hippocampal tissues of newborn mice in model group and control group; degeneration, reduction and enzymolysis of proteins were performed; iTRAQ reagent was used to label the proteins, LC - MS/MS was used to screen and identify differentially expressed pro- teins, biological analysis on differentially expressed proteins was conducted. Results： A total of 215 proteins were identified in hippocampal tissue of newborn mice, 158 proteins were differentially expressed, including 58 up- regulated proteins and 100 down- regulated proteins. These differentially expressed proteins were involved in 150 biological processes, 48 cellular components and 52 molecular functions. Protein interaction network diagram showed that Ctnnbl, Vcl and 14 -3 -3 protein family （Ywhag, Ywhae, Ywhah, Ywhaq, Ywhab） located in the main connected nodes. Conclusion： The proteins locating in the main connected nodes of protein interaction network diagram maybe play an important role in the process of brain development of newborn mice stimulated by fear during pregnancy.

关 键 词：ITRAQ 蛋白质组学 海马组织 脑发育 

分 类 号：R715[医药卫生—妇产科学]