丛生蛋白和Stathmin 2在癫痫患者及癫痫大鼠模型中的表达  

Altered expression of clusterin and stathmin2 in epileptic patients and rats

作　　者：苏宗翔 谢延风[1] 陆波[1] 陈丹[1] SU Zongxiang, XIE Yanfeng, LU Bo, CHEN Dan(Department of Neurosurgery, the First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China)

机构地区：[1]重庆医科大学附属第一医院神经外科,重庆400016

出　　处：《第三军医大学学报》2018年第19期1753-1761,共9页Journal of Third Military Medical University

基　　金：国家自然科学基金青年科学基金(81201002)~~

摘　　要：目的探讨丛生蛋白(Clusterin)和Stathmin2在癫痫患者及戊四氮点燃的癫痫大鼠模型中脑组织的表达变化,初步探讨Clusterin和Stathmin2在癫痫发生中的作用及机制。方法收集癫痫患者手术切除的皮质作为癫痫组(n=20);收集脑外伤患者手术切除的皮质作为对照组(n=20)。取100只SD雄性大鼠分为5组(EP0-4),每组20只。对照组(EP0组)大鼠未做任何处理,实验组(EP1-4组)大鼠隔天腹腔注射戊四氮,注射次数分别为1、5、10、15次。应用免疫荧光、免疫组化、Western blot检测和免疫共沉淀分析Clusterin和Stathmin2的定位、表达变化及相关性。结果①免疫荧光定位结果示Clusterin和Stathmin2主要分布于神经元的细胞膜和细胞质。②免疫组化和Western blot检测结果显示人癫痫组中Clusterin较对照组表达降低,而Stathmin2则表达升高,在大鼠实验组(EP1-4)及对照组(EP0)中也有同样趋势,且EP1-4组中Clusterin的表达随注射药物次数增加逐渐降低,Stathmin2表达则逐渐升高,以上所述差异均有统计学意义(P <0.05)。③免疫共沉淀结果示Clusterin与Stathmin2相互结合。结论 Clusterin和Stathmin2的结合及表达变化,可能参与到癫痫的发生、发展。ObjectiveTo explore the expression alterations of clusterin and stathmin2 in brain tissue of epileptic patients and rat model induced by pentylenetetrazol in order to investigate the roles and underlying mechanisms of the 2 molecules in the development of epilepsy. MethodsCortical samples of epileptic patients after surgery were collected as epileptic group （n=20）, and the samples from patients of cerebral trauma were collected as control group （n=20）. A total of 100 male SD rats were divided into 5 groups （EP04）, with 20 rats in each group. The rats in the control group （EP0） did not undergo any treatment. Those from the experimental groups （EP 1-4） were intraperitoneally injected with pentylenetetrazol every other day, with the total injection time of 1, 5, 10 and 15 times, respectively for each group. Immunofluorescence staining, immunohistochemical staining, Western blot analysis and coimmunoprecipitation were used to analyze the localization, expression changes and correlation of clusterin and stathmin2. Results①Immunofluorescence results showed that clusterin and stathmin2 were both located in the cell membrane and cytoplasm of the neurons. ②Immunohistochemical staining and Western blotting showed that the expression of clusterin in human epileptic group was decreased, while that of stathmin2 was increased. The same tendency was also observed in the experimental groups and control group of rats. In addition, the expression level of clusterin in the EP14 groups was gradually reduced, while that of stathmin2 was increased with the increase of injection time （both P〈0.05）. ③Co-immunoprecipitation showed that clusterin and stathmin2 were bound to each other. ConclusionBinding of clusterin to stathmin2 and their altered expression may be involved in the epileptogenesis.

关 键 词：癫痫 丛生蛋白 Stathmin2 戊四氮 

分 类 号：R394.3[医药卫生—医学遗传学] R742.1[医药卫生—基础医学]