【作者】 王倩；

【导师】 陈阳美；

【作者基本信息】 重庆医科大学 ， 神经病学， 2014， 硕士

【摘要】 目的：现研究认为树突棘的形态发生及数量的多少与苔藓纤维出芽及突触重塑密切相关。众多研究表明，具有调控活性的基因的转录表达，在神经环路的发生发展过程中起着关键的作用。p250GAP是一种Rho家族的GTP酶激活蛋白，其与多种突触蛋白相作用，水解下游底物上的GTP，从而启动底物相关的信号通路引起细胞骨架的解聚，减小树突棘的密度及体积。本研究主要通过检测癫痫患者和癫痫大鼠模型中p250GAP的表达，探讨其在癫痫发生发展过程中的作用与机制。方法：1.选择14例难治性癫痫患者颞叶手术切除标本和性别、年龄无差异的10例非癫痫患者颞叶组织标本。2.选择成年SD大鼠诱导氯化锂-匹鲁卡品模型，随机分为2组：正常对照组，癫痫模型组，根据癫痫进程分为6个亚组（24h，72h，7d，14d，30d，60d）。3.用westernblot技术和免疫组化技术检测p250GAP的表达水平。4.用免疫组化技术检测NPY表达水平。结果：1. p250GAP在颞叶癫痫患者及动物模型中表达均下降，在大鼠癫痫模型中在24h、72h、14d、30d、60d中p250GAP的表达水平明显低于正常对照组大鼠（P<0.05），7d时表达低于正常对照组（P>0.05），但差异无统计学意义。p250GAP在24h、14d、30d表达最低。2.免疫组化结果显示，在人标本及大鼠标本中p250GAP表达于神经元胞浆内，且TLE组及大鼠癫痫模型组, p250GAP阳性细胞数目及染色程度明显低于对照组；在癫痫大鼠中，NPY阳性细胞数目及染色程度明显高于正常对照，苔藓纤维出芽明显；Western blot提示p250GAP在TLE患者中,表达水平明显低于非TLE患者（P<0.05）;动物模型除7d组外（P>0.05），24h、72h、14d、30d、60d中p250GAP的表达水平明显低于正常对照组大鼠（P<0.05）。尤在1d、30d、60d时间点最低。结论：癫痫患者与癫痫大鼠模型中p250GAP表达均低于对照组，癫痫大鼠模型中p250GAP的表达在癫痫各期都降低，提示p250GAP可能与癫痫的发病及自发性发作密切相关。更多还原

【Abstract】 Objective Now research suggests that the morphology of dendriticspines and quantity was closely related to mossy fiber sprouting andSynaptic plasticity. Numerous studies have shown that activity-regulatedgene expression is believed to play a key role in the development andrefinement of neuronal circuitry. P250GAP which is a kind of Rho familyGTPase-activating protein interacted with a variety of synaptic protein tohydrolyze the GTP of downstream substrates. Then activated substraterelated signaling pathways that resulted in the depolymerization ofcytoskeleton and reduction of the density and volume of dendritic spines.This research mainly explore the role of p250GAP in pathogenesis ofepilepsy by detecting the expression of p250GAP in patients withintractable epilepsy and rat model of epilepsy.Methods1.14intractable epilepsy patients and10non-epileptic patients withno significantly differences on age and gender were included from which the temporal neocortex were surgically removed.2. Adult Sprague Dawley (SD) rats were selected to be Lithiumchloride-pilocarpine model, were randomly divided into2groups: normalcontrol group and epilepsy model group. Than according to the process ofepilepsy is divided into seven groups（24h，72h，7d，14d，30d，60d）3. Expression level of p250GAP was determinated by Western blottechnology and immunohistochemical technologyResults1. p250GAP in patients with temporal lobe epilepsy and epileptic ratmodel of each point in time were significantly reduced（P<0.05）.2. The immunohistochemical showed that p250GAP expression in thecytoplasm of neurons in the specimen and rats. Neuropeptide-Y(NPY)indicated mossy fiber sprouting(MFS) in model group.Western blot showedthat p250GAP expression level in TLE patients significantly lower thancontrol group (P <0.05); p250GAP expression level in model group waslower than control group,except7d， in the time point of24h,72h,7dand14d,30d,60d (P <0.05) and in the1d,30d,60d time pointsminimum。Conclusions P250GAP expression in people with epilepsy andepileptic rat model were lower than the control group.p250GAP expressionin epileptic rat model of incubation period and chronic phase were thelowest prompted p250GAP may be closely associated with the onset of epilepsy and epileptic seizures.更多还原