皮质发育障碍模型成年鼠海马蛋白质组学研究  

Proteomics research of hippocampus on rat with DCDs

作　　者：冯占辉 胡皓月 董楝 于云莉 晏宁 徐祖才 叶兰 FENG Zhanhui;HU Haoyue;DONG Lian(Department of Neurology, Affiliated Hosptial of Guizhou Medical University, Guiyang (550004) , Guizhou, China)

机构地区：[1]贵州医科大学附属医院神经内科,贵州贵阳550004 [2]重庆医科大学附属大学城医院神经内科 [3]遵义医科大学第一附属医院神经内科 [4]贵州医科大学基础医学院

出　　处：《癫痫与神经电生理学杂志》2019年第5期257-262,F0002,共7页Journal of Epileptology and Electroneurophysiology(China)

基　　金：国家自然科学基金(NO:81860248);贵阳市政府和贵州医科大学联合基金[NO:筑科合同(2017)30-32];中国抗癫痫协会癫痫基金(NO:2018016).

摘　　要：目的:利用双向凝胶电泳(2-DE)技术对皮质发育障碍(DCD)模型成年鼠海马与正常对照组海马的蛋白表达谱进行差异表达分析,获取与难治性癫痫密切相关的靶蛋白。方法:应用X射线175cGY剂量制作DCD模型鼠,从模型鼠和对照组中随机选择6只进入实验。将56 d大鼠断头取脑,剥离海马,提取总蛋白进行双向电泳,最终获得2-DE图谱,银染法对电泳后的凝胶染色,采集凝胶图像并进行图像分析,寻找差异蛋白点,对差异蛋白点进行酶解和质谱鉴定。结果:①PH3-10线性IPG胶条对各组蛋白质分析,成功获得分辨率高和重复性好的大鼠海马组织蛋白2-DE图谱;②经质谱鉴定最终获得8个差异蛋白点,GTP结合蛋白、突触相关蛋白90、突触囊泡蛋白和钙网蛋白等4个上调;肌酸激酶、Cu-Zn超氧化物歧化酶、二氢密噪相关蛋白和谷胱甘肽S-转移酶等4个下调。结论:放射线模型鼠海马蛋白质与对照组比较差异有显著意义,其中突触重塑和苔薛纤维芽生相关蛋白可能与癫痫易感性密切相关。Objective: To compare the difference of proteins of hippocampus between the animal model with disorders of cortical development (DCD) and the control and further find out the target and effective proteins related with intractable epilepsy. Methods: The animal model induced by 175cGy Xray was established? 6 rats were randomly selected in the model group and the control. The whole brain of each animal was rapidly dissected. The total protein of hippocampus was applied to obtain the 2-DE gel maps. Then, the gels were stained with silver and differential protein spots were analyzed by software analysis, subject to in-gel digestion, and identified by mass spectrometry. Results:①pH3-10 linearity IPG was applied to analyze protein and obtain high resolution and fine repetitiveness 2-DE gel images of rat brain proteome. cThrough the analysis of 2-DE image protein spots , eight different spots were obtained, four upregulated proteins: GTP-binding protein, synapse-associated protein 90, synaptotagmin 8 and calreticulin;four downregulated proteins: Cu-Zn superoxide dismutase, dihydropyrimidinase- related protein,creatine kinase and glutathione S-transferase P. Conclusion: The obvious expression difference of hippocampus protein between the rats with DCDs and the control were found. The protein related with the sprouting of mossy fiber and remolding of synapse may play an important role to potentiate epileptic activities.

关 键 词：皮质发育障碍(DCD) 成年大鼠 海马 蛋白质组学 

分 类 号：R741[医药卫生—神经病学与精神病学]