立体定向方法建立大鼠颞叶癫痫模型及其学习记忆功能障碍的实验研究

发布时间：2018-03-01 14:13

本文关键词： 癫痫颞叶/化学诱导迷宫学习记忆障碍/并发症钙离子　出处：《安徽医科大学》2006年硕士论文　论文类型：学位论文

【摘要】：目的：(1)探讨利用立体定向方法建立大鼠颞叶癫痫模型的方法以及观察发作后的脑电活动、MR表现和病理变化。(2)观察大鼠颞叶癫痫发作后不同时间段学习记忆功能障碍的情况以及探讨其与海马病理变化、海马神经元内游离钙离子浓度变化的相关性。方法：选择SD大鼠60只，随机分组法分为正常对照组、注射生理盐水组、注射红藻氨酸诱发颞叶癫痫发作后2周组、1月组、2月组、3月组，每组各10只。注射红藻氨酸组利用立体定向方法在大鼠右侧海马CA_3区注射红藻氨酸以诱发颞叶癫痫发作，注射生理盐水组在右侧海马CA_3区注射生理盐水，注射后分别用视频摄像机记录、观察大鼠行为学表现，定期在双侧海马、额叶皮质置入深部电极描记脑电变化。利用3.0T的MR对大鼠进行MRI和MRS检查。利用Morris水迷宫实验评价各组大鼠学习记忆障碍程度，随后处死大鼠进行病理检查，观察海马、额叶皮质的病理变化与学习记忆功能障碍的相关性。利用钙离子探针Fluo-3／AM，在激光扫描共聚焦显微镜下检测不同时间段的海马神经元内游离钙离子浓度的变化。结果：正常对照组和注射生理盐水组无颞叶癫痫发作和学习记忆障碍。右侧海马注射红藻氨酸的各组在注射红藻氨酸后均出现颞叶癫痫发作，脑电图描记出起源于注射侧海马并向对侧海马和双侧额叶皮质传导的癫痫波。MR检查提示注射红藻氨酸侧海马出现长TI长T2信号病灶。水迷宫实验中，有癫痫发作的各组出现不同程度的学习记忆功能障碍，定位航行实验中逃避潜伏期延长，空间搜索实验中在原有平台象限内游泳时间的百分比下降，与正常对照组和注射生理盐水组比较均有显著性差异(P＜0.05)，并随发作时间的延长逐渐加重，发作后2月达到高峰。病理检查发现，，注射红藻氨酸后右侧海马锥体细胞逐渐出现变性坏死，由CA_3区向CA_2、CA_4、CA_1区扩展，随后对侧海马锥体细胞也出现变性坏死，但程度明显低于注射侧，上述病理改变在注射后2月达到高峰，
[Abstract]:Objective: to explore the method of establishing rat temporal lobe epilepsy model by stereotactic method, and to observe the brain electrical activity and its pathological changes after seizure. 2) to observe the learning and memory impairment in different time periods after seizure of temporal lobe epilepsy in rats. The condition of obstruction and its relation to the pathological changes of the hippocampus, Methods: 60 SD rats were randomly divided into normal control group and saline injection group. Ten rats in each group were injected with kainic acid for 2 weeks, January, February and March, respectively. Kainic acid was injected into the CA_3 region of the right hippocampus of rats by stereotactic method to induce temporal lobe epilepsy. The rats in the saline group were injected with saline in the CA_3 area of the right hippocampus. The rats were recorded with video camera after injection, and the behavior of the rats was observed regularly in the bilateral hippocampus. MRI and MRS were examined by 3.0T Mr. The degree of learning and memory impairment was evaluated by Morris water maze test. Then the rats were sacrificed for pathological examination and hippocampal observation. The relationship between the pathological changes of frontal cortex and learning and memory impairment. Using calcium probe Fluo-3 / AM, the changes of free calcium concentration in hippocampal neurons at different periods of time were detected by laser scanning confocal microscope. There were no temporal lobe seizures and learning and memory disorders in normal control group and saline injection group. All the right hippocampal groups injected kainic acid had temporal lobe epilepsy after injection of kainic acid. EEG records of epileptic waves originating in the injected hippocampus and conducting to the contralateral hippocampus and bilateral frontal cortex suggest that the hippocampus injected with kainic acid presents a long TI long T2 signal lesion. Different degrees of learning and memory dysfunction appeared in each group with epileptic seizures, the latency of escape was prolonged, and the percentage of swimming time in the original platform quadrant decreased in the spatial search experiment. Compared with the normal control group and the saline injection group, there was significant difference (P < 0.05), and gradually aggravated with the prolongation of the attack time, and reached the peak on February after the attack. After injection of kainic acid, degeneration and necrosis of the right hippocampal pyramidal cells gradually appeared, which expanded from the CA_3 region to CA2CA4 and CA1 area, then the contralateral hippocampal pyramidal cells also showed degeneration and necrosis, but the degree was significantly lower than that of the injection side. The pathological changes peaked on February after injection.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R742.1;R-332

【引证文献】