吗啡成瘾和正常大鼠前额叶皮质差异表达蛋白的比较研究

THE PRIMARY COMPARATIVE STUDY ON CHANGES OF EXPRESSIVE PROTEOME OF PREFRONTAL CORTEX BETWEEN NORMAL AND MORPHINE-ADDICTED RATS

目的:比较吗啡成瘾与正常大鼠前额叶皮质(prefrontal cortex,PFC)蛋白质双向电泳图谱,寻找和鉴定吗啡成瘾大鼠PFC中的差异表达蛋白质。方法:以固相pH梯度等电聚焦为第一向和垂直SDS-PAGE为第二向,分别对正常对照大鼠和吗啡成瘾大鼠的PFC蛋白质样品进行二维分离,2-DE图谱经ImageMaster 2D Platinum v5.0软件分析,选取4个差异蛋白点用基质辅助激光解吸附离子化飞行时间质谱(MALDI-TOF-MS)进行鉴定。结果:通过对2-DE图谱蛋白斑点的匹配及对比分析,与吗啡成瘾相关的差异表达蛋白斑点为87个;经质谱鉴定出2个有意义的差异表达的蛋白斑点:snap25亚型β-snap25突触相关蛋白25、核不均一核糖核蛋白U。结论:吗啡成瘾组与正常对照组大鼠PFC蛋白质组存在差异;初步鉴定出了2种大鼠前额叶皮质中与吗啡成瘾相关的差异蛋白,其表达的变化可能通过多种途径影响PFC神经元功能,为研究阿片类物质依赖作用机制提出了新的思路和方向。

Objective：To obtain two-dimensional gel electrophoresis maps of the prefrontal cortex（PFC）proteins of normal and morphine-addicted rats for identifying the diferentially expressed proteins in the addicted rats.Methods： Rat models of morphine addiction were established,and the proteins in the PFC underwent two-dimensional gel electrophoresis with immobiline pH gradient isoelectric focusing as the first and vertica1 SDS-PAGE as the second dimension.Analysis of 2-DE maps was used to determine differential expression of proteins between the two groups by ImageMaster 2D Platinum v5.0,and four protein spots expressed differently were picked up for further identification by MALDI-TOF-MS.Results： Matched and compared with those of control group,87 protein spots were determined with differently expressive levels in morphine addiction group.By MALDI-TOF-MS,two protein spots of them were identified as β-snap25,an isoform of snap25（synaptosomal-associated protein 25）,and heterogeneous nuclear ribonucleoprotein U.Conclusion：There is obvious difference in expressive proteomes in PFC between normal and morphine-addicted rats.The functions of those identified proteins are involved in cell growth,apoptosis,differentiation,signal transduction,axon growth and cellular energy metabolism.Therefore proteomics can serve as a new approach in the study of morphine dependence to discover new therapeutic targets.