轴突起始段可塑性在MCD大鼠癫痫易感性中的作用研究

发布时间：2018-04-15 06:28

本文选题：皮质发育畸形 + 癫痫　；参考：《中南大学》2014年硕士论文

【摘要】：目的建立卡莫斯汀(BCNU)诱导的皮质发育畸形(malformation of cortical development, MCD)大鼠模型,观察MCD大鼠模型对氯化锂-匹罗卡品(LiCl-PILO)致痫敏感性,研究致痫后皮质神经元轴突起始段(axon initial segment, AIS)的可塑性变化规律,探讨AIS的可塑性变化在MCD大鼠癫痫易感性中的作用。方法 1.在孕鼠怀孕17天(E17d)时腹腔注射卡莫斯汀(BCNU),怀孕21天时生产仔鼠为MCD大鼠模型,Map1b免疫组化染色,验证MCD造模是否成功,取6-8周龄MCD大鼠和正常发育大鼠,腹腔注射LiCl-PILO诱导癫痫持续状态(status epileptics, SE),建立MCD癫痫大鼠模型； 2.记录和比较急性SE诱导成功率和死亡率,记录MCD和普通正常发育大鼠癫痫模型急性期和慢性期脑电图(EEG)； 3.免疫荧光研究AIS区可塑性：a.分组：MCD+SE组、MCD组、SE组和正常对照组；b.以SE后24小时、30天和60天为时间点取大鼠大脑组织制备冰冻切片,采用免疫荧光方法标记AIS区特异性蛋白ankyrinG,观察并测量各四组大鼠皮质感觉运动区神经元AIS区长度变化。结果 1.Map1b免疫组化染色提示成功建立了MCD模型。 2.MCD大鼠致痫成功率和总死亡率与正常发育大鼠无明显差异(P0.05),但致痫成功大鼠死亡率较高(P0.05)； 3.MCD大鼠和正常发育大鼠致痫急性期EEG形态相似,慢性期EEG可见自发放电,且自发放电幅度、持续时间、平均每小时自发放电次数均较正常发育大鼠增高(P0.001)； 4.皮质感觉运动区神经元AIS区长度：a.SE后24小时,MCD+SE组较SE组、MCD组显著缩短(P0.001),SE组较正常对照组亦明显缩短(P0.001),而MCD组与正常对照组相比无显著差异(P0.05); b.SE后30天和60天,MCD+SE组较MCD组明显缩短(P0.01和P0.001),SE组亦较正常对照组明显缩短,但MCD+SE组和SE组、MCD组和正常对照组之间无显著差异(P0.05)。结论MCD大鼠皮质感觉运动区神经元AIS区在致痫后出现缩短,结合脑电生理监测推断,AIS区形态与结构的可塑性改变可能参与了MCD大鼠模型对癫痫的易感。
[Abstract]:Objective to establish a rat model of cortical malformation of cortical development induced by carmosine (BCNU) and observe the sensitivity of MCD rat model to lithium-pilocarpine chloride (LiCl-PILO) induced epilepsy.To study the plasticity of axon initial segmenting (AIS) in the initial axon of post-epileptic cortical neurons, and to explore the role of the plasticity of AIS in the susceptibility to epilepsy in MCD rats.Method1.At the 17th day of gestation, carmosine was injected intraperitoneally into BCNU. On the 21st day of gestation, mice were produced as MCD rat model with Map1b immunohistochemical staining to verify the success of MCD model. 6-8 week-old MCD rats and normal developing rats were taken.The MCD epileptic model was established by intraperitoneal injection of LiCl-PILO status epileptics.2.The success rate and mortality rate of acute SE induction were recorded and compared. MCD and EEGG were recorded in the acute and chronic phase of epileptic model of normal developmental rats.3.The plasticity of AIS region was studied by immunofluorescence.The mice were divided into two groups: 1% MCD SE group, 2% MCD SE group and 2 normal control group.Frozen sections were prepared from rat brain tissue at 30 and 60 days after SE, and the length of AIS area of neurons in the sensory motor area of each group was observed and measured by immunofluorescence labeling of ankyrinG, a specific protein in the AIS region.Result1.Map1b immunohistochemical staining indicated that MCD model was successfully established.There was no significant difference in the success rate and the total mortality rate between the 2.MCD rats and the normal development rats (P 0.05), but the mortality rate of the rats with successful epilepsy was higher than that of the normal development rats (P 0.05).The morphology of EEG in 3.MCD rats was similar to that in normal developing rats. Spontaneous discharges were observed in chronic EEG, and the amplitude and duration of spontaneous discharges, the average number of spontaneous discharges per hour in 3.MCD rats were higher than those in normal developmental rats (P 0.001).4.The length of the AIS area of the cortical sensorimotor neurons was significantly shorter in the MCDSE group than that in the control group at 24 hours after 1: 00 a.SE, and in the MCD group, which was significantly shorter than that in the normal control group (P 0.05), and 30 and 60 days after b.SE.Compared with the MCD group, the SE group was significantly shorter than the MCD group, and the SE group was also significantly shorter than the normal control group.But there was no significant difference between MCD SE group and SE group and normal control group (P 0.05).Conclusion the cortical sensorimotor area AIS area of MCD rats is shortened after epilepsy. The plasticity of morphology and structure of MCD area may be involved in the susceptibility to epilepsy in MCD rat model combined with electrophysiological monitoring.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R742.1

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