p250GAP在难治性癫痫患者及癫痫动物模型中的表达及意义

发布时间：2018-02-27 17:37

本文关键词： 癫痫 p250GAP 树突棘细胞骨架苔藓纤维出芽　出处：《重庆医科大学》2014年硕士论文　论文类型：学位论文

【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（P0.05），7d时表达低于正常对照组（P0.05），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（P0.05）;动物模型除7d组外（P0.05），24h、72h、14d、30d、60d中p250GAP的表达水平明显低于正常对照组大鼠（P0.05）。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。
[Abstract]:Objective: this study suggests that the morphogenesis and number of dendritic spine are closely related to mossy fiber budding and synaptic remodeling. P250 gap is a kind of GTP enzyme activator of Rho family, which acts with many synaptic proteins. In this study, the expression of p250 gap in epileptic patients and epileptic rats was detected by hydrolysis of GTPs on downstream substrates, which activated the signal pathway associated with substrates to depolymerize cytoskeleton and reduce the density and volume of dendritic spine. To explore the role and mechanism of Epilepsy in the process of occurrence and development. Methods:. 1. The temporal lobe tissue specimens of 14 patients with intractable epilepsy and 10 patients with no difference in age were selected. 2. Adult SD rats were randomly divided into two groups: normal control group and epileptic model group. 3. The expression of p250 gap was detected by westernblot and immunohistochemistry. 4. Immunohistochemical technique was used to detect the expression of NPY. Results:. 1. The expression of p250 gap in patients with temporal lobe epilepsy and in animal models was decreased. The expression level of p250 gap in rat epileptic model was significantly lower than that in the control group at day 7 and day 7, but the difference was not statistically significant. The expression of p250 gap was the lowest at 24 h, 14 d and 30 d, but the expression level of p250 gap was lower than that of control group at day 7. The expression of p250 gap was lower than that of the control group at day 7, but the expression of p250 gap was lowest at 24 h, 14 d and 30 d. 2.Immunohistochemical results showed that p250 gap was expressed in the cytoplasm of neurons in human and rat specimens, and the number and staining degree of p250 gap positive cells in TLE group and epileptic model group were significantly lower than those in control group. The number and staining degree of NPY positive cells in epileptic rats were significantly higher than those in normal controls. Western blot suggested that p250 gap was present in TLE patients. The expression level of p250 gap was significantly lower in the animal model than that in the control group (P 0.05), and the expression level of p250 gap was significantly lower than that in the control group during the period of 60 days. The expression level of p250 gap was significantly lower than that in the control group, especially at the time point of 60 days after 1 day of 30 days. Conclusion: the expression of p250 gap in epileptic patients and epileptic rats is lower than that in the control group. The expression of p250 gap in epileptic rats is decreased in all stages of epilepsy, suggesting that p250 gap may be closely related to the onset and spontaneous seizure of epilepsy.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R742.1

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