丰富环境改善血管性痴呆大鼠学习和空间记忆能力及其机制研究
发布时间：2019-06-14 09:43
【摘要】：背景与目的:近年来由于人民群众生活质量的改善,心脑血管疾病的发生率也逐渐升高,由此导致血管性痴呆(VaD)的发病率也随之上升。目前在所有痴呆类型中,VaD发病率排名第二,仅低于阿尔茨海默病(AD),它在给病人带来生活上的困扰之外,也给社会和家庭造成了沉重的负担,目前临床上关于Va D病人的治疗方法仍十分有限。近年来,对于丰富环境(EE)的研究日益增多,发现其能增强神经的可塑性,在多种神经精神疾病中已经被证实有一定的治疗作用。在本研究中,我们拟探讨EE的干预是否能够提升Va D大鼠的学习及空间记忆能力以及其可能的内在作用机制。方法:本实验采用双侧颈总动脉结扎法(Bilateral common carotid occlusion,2VO)建立SD(Sprague-Dawley)大鼠的VaD模型。采用健康成年的雄性SD大鼠,随机分为4个组:假手术(sham)组、2VO组、假手术+丰富环境(sham+EE)组和2VO+丰富环境(2VO+EE)组。2VO组和2VO+EE组的大鼠进行2VO手术。有两种不同的饲养环境在本实验中被运用:标准饲养环境(SE)和丰富环境(EE)。Sham组和2VO组的大鼠在SE中饲养4周,而sham+EE组和2VO+EE组的大鼠则在EE中饲养4周。通过Morris水迷宫(MWM)来检测大鼠的学习及空间记忆能力的差异,尼氏染色观察大鼠神经元的存活数量及形态学的改变,tunel法测定各组大鼠神经细胞的凋亡情况,采用westernblot检测wnt/?-catenin通路中wnt3a、p-gsk-3?、?-catenin的蛋白含量的变化。结果:(1)在mwm的实验结果中,在ee中饲养的2vo+ee组大鼠的逃避潜伏期较se中饲养2vo组大鼠明显缩短(p0.05),目标象限停留时间明显增加(p0.05),在ee中饲养的sham+ee组大鼠逃避潜伏期较se中饲养的sham组大鼠明显缩短(p0.05),目标象限停留时间明显增加(p0.05),提示ee可以提高大鼠的学习和空间记忆能力。(2)与sham组相比,2vo组大鼠在术后4周的海马凋亡细胞比例明显增加(p0.05),而经过ee干预的2vo+ee组的海马凋亡细胞的比例则比2vo组明显下降(p0.05),同时经过ee干预的sham+ee组的海马凋亡细胞比例也较se中饲养的sham组大鼠明显下降(p0.05)。(3)与sham组相比,2vo组大鼠在术后4周的海马正常存活的神经元数量明显减少(p0.05)。而2vo+ee组在经过ee的干预后存活的神经元数量则2vo组增加(p0.05),经过ee干预的sham+ee组大鼠的海马存活神经元数量也明显要高于饲养在se环境中的sham组(p0.05)。(5)westernblot的结果显示,与se中饲养4周的sham组及2vo组大鼠相比,ee中饲养4周的sham+ee组和2vo+ee组大鼠海马中wnt3a、p-gsk-3?、?-catenin蛋白的含量明显增加(p0.05),提示大鼠在经历丰富环境的过程中伴随着wnt/?-catenin信号通路的激活。结论:与SE中饲养的大鼠相比,经历4周EE的大鼠学习和空间记忆能力得到了明显的提升。大鼠在EE的饲养环境下,海马神经元凋亡比例降低,正常神经元的数量增加。在EE的作用过程中同时伴随着wnt3a、p-GSK-3?、?-catenin蛋白含量的增加,说明4周的EE干预激活了wnt/?-catenin信号通路,提示wnt/?-catenin信号通路的激活可能是EE促进学习和空间记忆能力提升作用的内在分子学机制之一。
[Abstract]:BACKGROUND & OBJECTIVE: In recent years, the incidence of cardiovascular and cerebrovascular diseases has increased gradually due to the improvement of the quality of people's life, and the incidence of vascular dementia (VaD) has also increased. In all the types of dementia, the rate of VaD is the second and only lower than the Alzheimer's disease (AD), which causes a heavy burden on the society and the family, which is still very limited in the treatment of the Va-D patients. In recent years, the research on the rich environment (EE) has been increasing, and it has been found that it can enhance the plasticity of the nerve, and has been proved to have a certain therapeutic effect in various neuropsychiatric diseases. In this study, we intend to explore whether the intervention of EE can improve the learning and spatial memory capacity of the Va-D rats and its possible internal mechanism of action. Methods: The VaD model of SD (Sprague-Dawley) rats was established by the double-sided common carotid artery ligation (2VO). Healthy adult male SD rats were randomly divided into 4 groups: sham group, 2VO group, sham operation + rich environment (sham + EE) group and 2VO + rich environment (2VO + EE) group. Two different breeding environments were used in this experiment: the standard feeding environment (SE) and the rich environment (EE). Rats in the Sham group and the 2 VO group were fed in SE for 4 weeks, while the sham + EE group and the 2 VO + EE group rats were housed in the EE for 4 weeks. The learning and spatial memory abilities of the rats were detected by the Morris water maze (MWM). The number of survival and the morphological changes of the neurons in the rats were observed by means of the method of Morris water maze (MWM), and the apoptosis of the nerve cells in each group was determined by means of the test. The wnt3a, p-gsk-3 in the wnt/?-cattenin pathway were detected by using the method of western blot. ? -a change in the protein content of catenin. Results: (1) In the experimental results of mwm, the escape latency of the 2 vo + ee group rats fed in ee was significantly shortened (p0.05), and the retention time of the target quadrant was significantly increased (p0.05). In the sham + ee group, the rats of sham + ee group were significantly shorter (p0.05) than that of the sham group, and the retention time of the target quadrant was significantly increased (p0.05), and it was suggested that the ee could improve the learning and spatial memory of the rats. (2) Compared with the sham group, the proportion of the apoptosis cells in the hippocampus of the 2vo + ee group after the operation was significantly increased (p0.05), while the proportion of the apoptotic cells in the hippocampus of the 2vo + ee group after the ee intervention was significantly lower than that of the second group (p0.05). At the same time, the percentage of apoptotic cells in the hippocampus of the sham + ee group by ee intervention was lower than that in the sham group (p0.05). (3) Compared with the sham group, the number of neurons in the 2 vo group in the normal survival of the hippocampus at 4 weeks after operation was significantly reduced (p0.05). The number of surviving neurons in the 2vo + ee group after the ee intervention increased (p0.05), and the number of surviving neurons in the hippocampus of the sham + ee group after the ee intervention was significantly higher than that in the sham group (p0.05) in the se environment. (5) The results of western blot showed that wnt3a, p-gsk-3 in the hippocampus of the 2 vo + ee group and the 2 vo + ee group were fed with 4 weeks of sham + ee and 2 vo + ee groups in the ee. ? The content of the-catenin protein was significantly increased (p0.05), suggesting that the rats were associated with the activation of the wnt/?-cattenin signal pathway in a rich environment. Conclusion: Compared with the rats fed with SE, the learning and spatial memory ability of rats subjected to 4 weeks of EE is obviously improved. In the environment of EE, the percentage of neuron apoptosis in the hippocampus decreased and the number of normal neurons increased. At the same time with wnt3a, p-GSK-3 in the course of the action of EE? ? The increase in the content of the-catenin protein suggests that the 4-week EE intervention has activated the wnt/?-cattenin signal pathway, suggesting that the activation of the wnt/?-catenin signal pathway may be one of the intrinsic molecular mechanisms of EE for learning and the enhancement of spatial memory capacity.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R749.1
本文编号：2499299
[Abstract]:BACKGROUND & OBJECTIVE: In recent years, the incidence of cardiovascular and cerebrovascular diseases has increased gradually due to the improvement of the quality of people's life, and the incidence of vascular dementia (VaD) has also increased. In all the types of dementia, the rate of VaD is the second and only lower than the Alzheimer's disease (AD), which causes a heavy burden on the society and the family, which is still very limited in the treatment of the Va-D patients. In recent years, the research on the rich environment (EE) has been increasing, and it has been found that it can enhance the plasticity of the nerve, and has been proved to have a certain therapeutic effect in various neuropsychiatric diseases. In this study, we intend to explore whether the intervention of EE can improve the learning and spatial memory capacity of the Va-D rats and its possible internal mechanism of action. Methods: The VaD model of SD (Sprague-Dawley) rats was established by the double-sided common carotid artery ligation (2VO). Healthy adult male SD rats were randomly divided into 4 groups: sham group, 2VO group, sham operation + rich environment (sham + EE) group and 2VO + rich environment (2VO + EE) group. Two different breeding environments were used in this experiment: the standard feeding environment (SE) and the rich environment (EE). Rats in the Sham group and the 2 VO group were fed in SE for 4 weeks, while the sham + EE group and the 2 VO + EE group rats were housed in the EE for 4 weeks. The learning and spatial memory abilities of the rats were detected by the Morris water maze (MWM). The number of survival and the morphological changes of the neurons in the rats were observed by means of the method of Morris water maze (MWM), and the apoptosis of the nerve cells in each group was determined by means of the test. The wnt3a, p-gsk-3 in the wnt/?-cattenin pathway were detected by using the method of western blot. ? -a change in the protein content of catenin. Results: (1) In the experimental results of mwm, the escape latency of the 2 vo + ee group rats fed in ee was significantly shortened (p0.05), and the retention time of the target quadrant was significantly increased (p0.05). In the sham + ee group, the rats of sham + ee group were significantly shorter (p0.05) than that of the sham group, and the retention time of the target quadrant was significantly increased (p0.05), and it was suggested that the ee could improve the learning and spatial memory of the rats. (2) Compared with the sham group, the proportion of the apoptosis cells in the hippocampus of the 2vo + ee group after the operation was significantly increased (p0.05), while the proportion of the apoptotic cells in the hippocampus of the 2vo + ee group after the ee intervention was significantly lower than that of the second group (p0.05). At the same time, the percentage of apoptotic cells in the hippocampus of the sham + ee group by ee intervention was lower than that in the sham group (p0.05). (3) Compared with the sham group, the number of neurons in the 2 vo group in the normal survival of the hippocampus at 4 weeks after operation was significantly reduced (p0.05). The number of surviving neurons in the 2vo + ee group after the ee intervention increased (p0.05), and the number of surviving neurons in the hippocampus of the sham + ee group after the ee intervention was significantly higher than that in the sham group (p0.05) in the se environment. (5) The results of western blot showed that wnt3a, p-gsk-3 in the hippocampus of the 2 vo + ee group and the 2 vo + ee group were fed with 4 weeks of sham + ee and 2 vo + ee groups in the ee. ? The content of the-catenin protein was significantly increased (p0.05), suggesting that the rats were associated with the activation of the wnt/?-cattenin signal pathway in a rich environment. Conclusion: Compared with the rats fed with SE, the learning and spatial memory ability of rats subjected to 4 weeks of EE is obviously improved. In the environment of EE, the percentage of neuron apoptosis in the hippocampus decreased and the number of normal neurons increased. At the same time with wnt3a, p-GSK-3 in the course of the action of EE? ? The increase in the content of the-catenin protein suggests that the 4-week EE intervention has activated the wnt/?-cattenin signal pathway, suggesting that the activation of the wnt/?-catenin signal pathway may be one of the intrinsic molecular mechanisms of EE for learning and the enhancement of spatial memory capacity.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R749.1
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