海人酸诱导癫痫大鼠的海马微血管的改变

发布时间：2018-03-06 16:33

本文选题：癫痫　切入点：海人酸　出处：《大连医科大学》2007年硕士论文　论文类型：学位论文

【摘要】： 背景:癫痫(epilepsy)是由多种病因引起的脑功能障碍综合征,是脑细胞群异常的超同步放电而引起的发作性、突然的、暂时性的脑功能紊乱。海人酸(KA)是脑内兴奋性氨基酸递质—谷氨酸的结构类似物。海人酸模型已广泛用于癫痫发作的研究。海马是脑内对最缺血敏感的区域之一。上世纪初Scharrer等就指出海马结构的易损性主要归因于其血液供应的独特方式。辜清等已在扫描电镜下观察到听源性癫痫大鼠海马内存在微血管病变。目的:,本文采用碱性磷酸酶法对KA癫痫模型中海马CA1区的微血管分布密度进行定量观察,以探求海马微血管构筑在颞叶癫痫发病机制中的作用,为癫痫发病机制研究提供基础资料。方法:⑴雄性SD大鼠12只(180～250g),分为实验组(KA, n=6)和对照组(NS, n=6)。⑵实验组大鼠颈部皮下注射KA(10mg/kg),根据Racine[15]所描述的癫痫行为1～5级标准,观察实验组动物全部达到4级或5级后,即认为实验组动物已符合癫痫模型的要求;对照组大鼠颈部皮下注射生理盐水。⑶对照组和实验组动物分别在给予生理盐水或KA7d后,水合氯醛麻醉,心脏灌流固定,取脑,后固定,酒精梯度脱水,火棉胶包埋,切片(厚90μm),碱性磷酸酶染色,常规封片。⑷光镜观察,使用Nikon microscope picture analysis system分析软件进行定量分析。取右侧背侧海马最大的部位,连续观察3张切片,目标区域为CA1区分子层,从CA1区的内侧端开始连续计数5个高倍视野(36300×90μm~3 ),记录每个视野的微血管数目(N)、微血管最大直径的平均值(D)和视野内血管的总长度(L)。计算各指标的平均值和微血管的平均长度(L/N)。⑸结果用均数±标准差表示;应用spss11.5软件的单因素方差分析(one-way ANOVA)进行统计;P0.05具有统计学意义。结果:⑴海马内微血管的分布具有与神经组织相对应的层次性:区各层之间微血管密度及分布的层次性最明显,其中多形层微血管最丰富,锥体层的最稀少,分子层的较稀少;CA3区中的锥体层血管密度最高,多形层次之,分子层最稀。CA4区的微血管分布没有明显的层次性。齿状回的微血管分布以分子层最密,多形层次之,颗粒层最稀。对照组和实验组大鼠海马微血管的在分布方式上未见差别。⑵实验组血管数目为5.21±0.62,明显高于对照组(3.56±0.76), P=0.001;实验组血管平均直径为4.25±0.23μm,明显低于对照组(5.35±1.04μm),P=0.030;实验组血管长度为828.81±82.75μm ,明显长于对照组( 438.12±64.23μm),P=0.000;实验组血管的平均长度为198.23±24.48μm,略高于对照组(151.77±54.25μm),但不具有显著性,P=0.085。结论:⑴海马内微血管的分布具有与神经组织相对应的层次性,癫痫大鼠海马微血管构筑保持了层次性特点。⑵大鼠海马CA1区分子层的微血管密度在KA大鼠癫痫发作7天后发生明显增加。
[Abstract]:Background: epilepsy epilepsyis is a syndrome of brain dysfunction caused by various causes. It is a sudden and paroxysmal disorder caused by abnormal supersonic discharges of brain cell groups. Kainic acid is a structural analogue of excitatory amino acid transmitter-glutamic acid in the brain. The kainic acid model has been widely used in the study of epileptic seizures. The hippocampus is one of the most ischemic sensitive regions of the brain. At the beginning of last century, Scharrer et al pointed out that the vulnerability of hippocampal formation was mainly attributed to the unique way of blood supply. Koo Qing et al had observed microangiopathy in hippocampus of auditory epileptic rats under scanning electron microscope. Objective to investigate the role of hippocampal microvascular architecture in the pathogenesis of temporal lobe epilepsy by quantitative observation of microvessel density in hippocampal CA1 region in Ka epileptic model by alkaline phosphatase method. To provide basic data for the study of the pathogenesis of epilepsy. Methods Twelve male Sprague-Dawley rats (n = 12) were divided into two groups: the experimental group (K A, n = 6) and the control group (n = 6.2). The rats of the experimental group were subcutaneously injected with 10 mg 路kg ~ (-1) 路kg ~ (-1) of Kar. According to the standard of epilepsy described by Racine [15], the animals in the experimental group were all observed to reach grade 4 or grade 5. That is to say, the experimental group has met the requirements of epileptic model, the control group rats were injected with normal saline 3.3. the control group and the experimental group were given normal saline or KA7d respectively, chloral hydrate was anesthetized, the heart was perfused and fixed, and the brain was taken out. Posterior fixation, alcohol gradient dehydration, colloid embedding, slice (90 渭 m thick, alkaline phosphatase staining, routine 4 light microscope observation), quantitative analysis using Nikon microscope picture analysis system software. The largest part of the right dorsal hippocampus was taken. Three slices were observed continuously, and the target region was the molecular layer of CA1 region. Starting from the medial end of the CA1 region, 5 high power visual fields 36 300 脳 90 渭 m were counted continuously. The number of microvessels in each field was recorded, the mean value of the maximum diameter of the microvessels (D) and the total length of the vessels in the field of vision were recorded. The mean values of each index and the microvessels were calculated. The mean length of L / N ~ (5) is expressed as mean 卤standard deviation. Using spss11.5 software one-way ANOVA (one-way ANOVA) was statistically significant. Results the distribution of microvessels in the hippocampus of 1 / 1 had the same level as that of the nerve tissue: the microvessel density and distribution between the layers of the region were the most obvious, in which the polymorphic layer was the most abundant, and the pyramidal layer was the most rare. The density of blood vessels in the pyramidal layer of the molecular layer is the highest and the polymorphic layer of the pyramidal layer is the highest in the molecular layer. There is no obvious stratification of the microvessel distribution in the most thinnest part of the molecular layer. The microvessel distribution in the dentate gyrus is the densest and polymorphic layer in the dentate gyrus. There was no difference in the distribution of hippocampal microvessels between the control group and the experimental group (5.21 卤0.62), which was significantly higher than that of the control group (3.56 卤0.76), P0. 001, and the mean diameter of the experimental group was 4.25 卤0.23 渭 m, which was significantly lower than that of the control group (5.35 卤1.04 渭 m). The vascular length of the test group was 828.81 卤82.75 渭 m. It was significantly longer than that in the control group (438.12 卤64.23 渭 m P0. 000), and the average vascular length in the experimental group was 198.23 卤24.48 渭 m, which was slightly higher than that in the control group (151.77 卤54.25 渭 m), but not in the control group (P 0.085). Conclusion the distribution of microvessels in the hippocampus of 1 / 1 has a hierarchy corresponding to that of the nerve tissue. Hippocampal microvascular architecture in epileptic rats maintained hierarchical characteristics. 2 the microvessel density in the molecular layer of hippocampal CA1 region in rats increased significantly 7 days after seizure in Ka rats.
【学位授予单位】：大连医科大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R742.1;R361

【参考文献】