皮质发育不良大鼠差异蛋白的筛选及其致痫作用的探讨  

Comparative proteomics of rat brain in the BCNU-induced model of cortical dysplasia

作　　者：郭谊[1] 张曼曼[1] 丁瑶[1] 杨怡[1] 蒋艳[1] 呼建文 丁美萍[1] 

机构地区：[1]浙江大学医学院附属第二医院神经内科,杭州310009 [2]中科新生命生物科技有限公司

出　　处：《中华急诊医学杂志》2013年第12期1355-1359,共5页Chinese Journal of Emergency Medicine

基　　金：浙江省自然科学基金（Y2080475）;国家自然科学基金（81000556、81171227）

摘　　要：目的寻找皮质发育不良大鼠脑组织（皮质和海马）与正常对照组织在不同发育阶段的差异表达蛋白质，探讨这些蛋白质在皮质发育不良所致癫痫发生机制中的作用。方法妊娠晚期（孕17d）Sprague—Dawley大鼠腹腔注射卡莫司汀（15mg／kg）诱导仔鼠皮质发育不良。提取新生期（出生后7d，记作fy7）及成年仔鼠（P60）新皮质及海马组织蛋白，运用双向凝胶电泳技术构建卡莫司汀诱导皮质发育不良鼠和正常鼠皮质及海马组织在新生期及成年期的蛋白质表达谱，通过比对分析筛选出差异表达的蛋白质斑点。应用MALDI—TOF—MS技术平台对差异最为显著的部分蛋白进行鉴定。结果在模型组和对照组之间共筛选出57个差异表达蛋白质斑点（P〈0．05，35个来自新生期，22个来自成年期），其中35个在模型组表达上调，22个在模型组表达下调。12个蛋白质斑点得以成功鉴定，包括微管蛋白、肌动蛋白、胶质纤维酸性蛋白、生长相关蛋白-43、广泛型线粒体肌酸激酶、甘油醛-3-磷酸脱氢酶和脑型原肌球蛋白等蛋白或其亚单位线粒体肌酸激酶广泛型亚型。结论鉴定的蛋白质参与细胞骨架构成、突触形成和线粒体功能活动等多种生理过程，它们的表达改变可能在皮质发育不良致癫痫发牛的机制中发挥重要作用。Objective To screen the differential proteins in the brain （neocortex and hippocampus） between the rats with cortical dysplasia （CD） and control ones, and investigate the role of their alteration in the development of epilepsy in CD. Methods Cortical dysplasia was induced in rat pups via in utero delivery of BCNU. A two-dimensional eleetrophoresis （2-DE） -based approach was used to construct the expression profiles of proteins in both the neocortex and hippocampns at different age groups （ postnatal day 7 and 60） and to detect proteome changes between CD rats and control ones. Following gel image analysis, protein spots that differed in abundance between CD and control rats were identified by using Matrx-assisted laser desorption/ionization （MALDI） mass spectrometry （MS） and MS/MS. Results A total of 57 kinds of protein were screened out （ P 〈 0. 05 ） , in which 35 were found up-regulated and 22 were down-regulated compared with the control, 35 from neonatal stage （postnatal day 7 ） and others from adult stage. Finally, 12 among them were identified, including tubulin, alpha-lB, Beta-actin, tubulin beta-2A, GAP-43, UbCKmit, GAPDH and TMBr-3, etc. Conclusions Changed expression of specific proteins which werefound in our study are involved in construction of brain ＂s cytoskeleton, synaptic function, mitochondrial function and so forth. Thus, they may be related to the pathogenic mechanisms of epileptogenieity of CD.

关 键 词：皮质发育不良 卡莫司汀 新皮质 海马 双向电泳 质谱 差异蛋白表达 癫痫 

分 类 号：R742.1[医药卫生—神经病学与精神病学]