吗啡成瘾和正常大鼠的前额叶皮质表达蛋白质组变化的初步研究

The primary study on changes of expressive proteome of the prefrontal cortex between normal and morphine-addicted rats

目的建立和比较吗啡成瘾与正常大鼠前额叶皮质(PFC)蛋白质双向电泳图谱,寻找和鉴定吗啡成瘾大鼠PFC中的差异蛋白表达谱。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 Platinumv5.0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、β-肌动蛋白。结论吗啡成瘾组与正常对照组大鼠PFC蛋白表达存在差异;初步鉴定了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经影响PFC神经元功能,为我们研究阿片类物质依赖作用机制提出了新的思路和方向。

Aim 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.The proteins in the PFC underwent two-dimensiona1 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 expressions 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 had been determined with differently expressive levels in morphine addiction group.By MALDI-TOF-MS,two protein spots of them had been identified as Snap25 Isoform β-Snap 25 of Synaptosomal-associated protein 25 and Actb Actin,cytoplasmic 1.Conclusions 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,so proteomics can serve as a new approach in the study of morphine dependence to discover new therapeutic targets.