丰富环境对HIBD新生大鼠学习记忆、海马区BDNF和Synaptophys蛋白表达的影响

发布时间：2018-03-03 21:38

本文选题：丰富环境　切入点：缺氧缺血性脑损伤　出处：《青岛大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的本研究拟通过制作7日龄(相当于人类新生儿期)缺氧缺血性脑损伤(hypoxic-ischemic brain damage,HIBD)新生大鼠模型,早期给予不同强度丰富环境(environmental enrichment,EE)干预,动态观察在不同干预强度下HIBD大鼠学习记忆能力、海马区突触可塑性相关分子(脑源性神经营养因子、突触素蛋白)的表达变化,以探讨HIBD大鼠早期丰富环境干预的有效性及机制,尤其是不同干预强度对HIBD大鼠学习记忆功能、海马突触可塑性的影响及其分子机制,从而为临床HIBD患儿丰富环境的构建及康复方案的制定提供可能的理论依据,指导临床实践。方法将40只7日龄Wistar雄性大鼠随机分为以下四组:6小时丰富环境干预组(6h EE)、12小时丰富环境干预组(12h EE)、模型组(Model)和假手术组(Sham),每组均10只。丰富环境干预组和模型组均运用Rice法(1981)制备大鼠缺氧缺血脑损伤模型。丰富环境干预组分别进行14天的6h和12h丰富环境干预。干预结束后运用Morris水迷宫行为学实验测查大鼠的学习记忆能力。运用免疫印迹(Western blot)法检测大鼠海马区脑源性神经营养因子(BDNF)、突触素蛋白的表达。结果1、水迷宫实验的定位航行实验中,4组大鼠训练期间的逃避潜伏期随着训练天数的增加逐渐降低,但4组大鼠之间的逃避潜伏期比较差异无统计学意义(F=0.2369,P0.05)。2、水迷宫实验中,第5天空间探索测试时,模型组在目标象限寻找平台的时间与在其他象限比较,差异无统计学意义(P0.05);6h EE组、12h EE组、假手术组在目标象限寻找平台的时间显著长于其他象限,差异有统计学意义(P0.05)。3、水迷宫实验中,第7天空间探索测试时,4组大鼠在目标象限寻找平台的时间都长于其他象限,说明4组大鼠均能记住平台所在象限位置。6h丰富环境干预组(((?)±s)6h EE=38.132±2.876,P0.001)、12h丰富环境干预组(((?)±s)12h EE=45.526±2.648,P0.001)和模型组(((?)±s)_(Model)=30.702±3.819,P0.001)的新生大鼠在目标象限寻找平台的时间均短于假手术组(54.400±3.152),并且海马区BDNF(((?)±s)6h EE=0.529±4、0.038,P0.001;((?)±s)12h EE=0.660±0.034,P0.01;((?)±s)_(Model)=0.225±0.015,P0.001)和突触素蛋白(((?)±s)6h EE=0.889±0.027,P0.001;((?)±s)12h EE=1.114±0.037,P0.05;((?)±s)_(Model)=0.672±0.057,P0.001)的表达相比于假手术组(((?)±s)BDNF=0.803±0.026;5、((?)±s)突触素=1.347±0.092)也有所下降。6、与模型组相比,6h丰富环境干预组和12h丰富环境干预组的新生大鼠在目标象限寻找平台的时间更长(P6h EE0.01;P12h EE0.001),并且海马区BDNF(P6h EE0.001;P12h EE0.001)和突触素蛋白(P6h EE0.05;P12h EE0.001)的表达显著上升。7、更值得提及的是12h丰富环境干预组在目标象限寻找平台的时间(P0.01)以及海马区BDNF(P0.01)与突触素蛋白(P0.05)的表达情况都优于6h丰富环境干预组。结论丰富环境能够改善缺氧缺血性脑损伤新生大鼠的学习记忆能力,保护神经元细胞,其机制可能与上调海马区BDNF和突触素蛋白表达有关。
[Abstract]:Objective to establish a neonatal rat model of hypoxic-ischemic brain damage (HIBD) with hypoxic-ischemic brain damage (HIBD) at 7 days of age (equivalent to human newborn), and to give early intervention with environmental hazards of different intensities. The learning and memory ability of HIBD rats and the expression of synaptic plasticity related molecules (brain-derived neurotrophic factor, synaptophysin protein) in hippocampus were observed dynamically under different intervention intensity. To explore the effectiveness and mechanism of early rich environment intervention in HIBD rats, especially the effects of different intervention intensities on learning and memory function, hippocampal synaptic plasticity and its molecular mechanism in HIBD rats. Thus, it can provide a theoretical basis for the construction of rich environment and the establishment of rehabilitation program for children with HIBD. Methods 40 7-day-old Wistar male rats were randomly divided into the following four groups: 6 hour enriched environment intervention group (6 hours) and 12 hour enriched environment intervention group (12 h EEN, model group) and sham operation group (10 rats in each group). The model of hypoxic-ischemic brain injury in rats was established by Rice method in both the intervention group and the model group. The rich environment intervention group was treated with 14 days of rich environment intervention for 6 hours and 12 hours, respectively. After the intervention, Morris water maze test was used to test the behavior of rats. The expression of brain-derived neurotrophic factor (BDNF) and synaptophysin (synaptophysin) in the hippocampus of rats was detected by Western blot method. The escape latency decreased with the increase of training days. However, there was no significant difference in the escape latency among the four groups. In the water maze experiment, the time of finding the platform in the target quadrant in the model group was compared with that in the other quadrants during the space exploration test on the 5th day in the water maze experiment. There was no significant difference in the time of searching platform in the target quadrant between the sham operation group and the other quadrants. The difference was statistically significant (P 0.05). 3. In the water maze experiment, there was no significant difference between the two groups. On the 7th day of space exploration test, the time of searching platform in the target quadrant was longer in the 4 groups than in other quadrants, indicating that the four groups could remember the location of the platform in the quadrant. EE=38.132 卤2.876% P0.001 + 12h enriched environmental intervention group (EE=38.132 卤2.876 P0.001) for 12 hours. ) 12 h EE=45.526 卤2.648 (P0.001) and model group (P 0.001). The time of finding platform in the target quadrant of the neonatal rats was shorter than that of the sham-operated group (54.400 卤3.152) and the hippocampal area of BDNF? EE=0.529 卤4 0. 038? P0. 001? ) 12 h EE=0.660 卤0.034 P0.01? ) and synaptophysin? 6 h EE=0.889 卤0.027 P0.001? ) 12 h EE=1.114 卤0.037 P0.05? Compared with sham operation group (P 0.001), the expression of 0.672 卤0.057 (P 0.001) was significantly higher than that of sham operation group (P < 0.05). ) BDNF 0. 803 卤0. 026? ) 卤s) synaptophysin 1. 347 卤0. 092) was also decreased. Compared with the model group, the neonatal rats in the 6 h enriched environment intervention group and the 12 hour enriched environment intervention group had longer time searching for platform in the target quadrant, and the hippocampal area BDNF(P6h EE0. 001 P12 h EE0. 001) and synaptophysin protein. The expression of BDNFP0.01) and synaptophysin protein (P0.05) in hippocampus of 12h enriched environment intervention group were significantly higher than that of 6h rich environment intervention group (P0.01), and the expression of synaptophysin protein P0.05) was better than that of 6-hour enriched environment intervention group. Rich environment can improve the learning and memory ability of neonatal rats with hypoxic-ischemic brain injury. The protective mechanism of neuronal cells may be related to the up-regulation of BDNF and synaptophysin expression in hippocampal area.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R742

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