Ephrin-B3在癫痫大鼠海马中的动态表达变化  

Dynamic changes of Ephrin-B3 expression in hippocampus of a rat model of temporal lobe epilepsy

作　　者：刘田田 滕军放[1] 刘恒方[2] 张敏[2] 肖波[3] LIU Tiantian;TENG Junfang;LIU Hengfang(Department of Neurology,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China)

机构地区：[1]郑州大学第一附属医院神经内科,河南郑州450052 [2]郑州大学第五附属医院神经内科,河南郑州450054 [3]中南大学湘雅医院神经内科,湖南长沙410008

出　　处：《中风与神经疾病杂志》2021年第8期714-717,共4页Journal of Apoplexy and Nervous Diseases

基　　金：国家自然科学基金项目(No.81901326);河南省高等学校重点科研项目计划(19A320038);河南省医学科技攻关计划联合共建项目(2018020066)。

摘　　要：目的检测癫痫大鼠模型Ephrin-B3的动态表达,探讨其在颞叶癫痫中致痫作用。方法SD雄性大鼠随机分成对照组和实验组,构建氯化锂-匹罗卡品癫痫大鼠模型,在癫痫持续状态(SE)诱导成功后7 d(急性期)、14 d(静止期)、60 d(慢性期)分别采用Real-time qPCR、Western blot及免疫组化方法检测大鼠海马齿状回Ephrin-B3 mRNA及蛋白的表达及分布。结果在SE后的急性期和静止期,实验组大鼠均未见自发痫性发作;慢性期实验组大鼠有节律性点头、咀嚼、单侧或双侧前肢阵挛发作。荧光定量PCR结果显示:与对照组比较,急性期癫痫大鼠海马的Ephrin-B3 mRNA表达水平下降,静止期下降最为明显,慢性期有所回升,但仍低于对照组(P<0.001)。Western blot检测Ephrin-B3蛋白的表达水平与mRNA表达趋势相一致:与对照组比较,致痫后7 d、14 d、60 d的Ephrin-B3蛋白表达均下调,差异有统计学意义(P<0.001)。免疫组化结果显示,Ephrin-B3蛋白主要分布于海马齿状回的颗粒细胞胞膜上,其光密度值与mRNA的变化趋势相一致(P<0.001)。结论Ephrin-B3在海马组织的动态变化可能与慢性自发痫性发作密切相关。Objective To explore dynamic expression of Ephrin-B3 in hippocampus of pilocarpine induced epileptic rats and the role of Ephrin-B3 in the process of epileptogenesis.Methods We first established pilocarpine induced epileptic rat model and then detected Ephrin-B3 expression at 7,14 and 60 days post status epilepticus(SE)by real time qPCR,immunohistochemistry and Western blot.Results After pilocarpine injection,rats showed facial muscles,forelimb and tail clonus and ataxic lurching,head bobbing and wet dog shakes.Real time qPCR revealed that Ephrin-B3 mRNA expression in hippocampus decreased during spontaneous seizure period.Compared with controls,epileptic rats showed decreased Ephrin-B3 expression at 7 days,reached the lowest level at 14 days and then increased slightly at 60 days post-SE(P<0.001).Ephrin-B3 protein in hippocampus also decreased at 7,14 and 60 days detected by Western blot(P<0.001).Immunohistochemistry results showed that Ephrin-B3 mainly expressed in granule cells in dentate gyrus.Analysis of immunohistochemistry revealed decreased Ephrin-B3 expression in epileptic rats compared with controls(P<0.001).Conclusion The dynamic changes of Ephrin-B3 expression in hippocampus of epileptic rats indicated that Ephrin-B3 may participate in the process of spontaneous recurrent seizures.

关 键 词：Ephrin-B3 癫痫 海马齿状回 慢性自发痫性发作 

分 类 号：R742.1[医药卫生—神经病学与精神病学]