慢性脑缺血大鼠海马区Numb表达变化的研究
发布时间：2018-07-25 15:17
【摘要】：研究背景及目的 随着人类社会老龄化的进程,痴呆的发病率也逐年增加。在痴呆人群中血管性痴呆(VD)和阿尔兹海默病(AD)占各类痴呆发病的绝大部分,其中VD发病率略高于AD。给社会和家庭带来了严重的经济和精神负担。近年来认为无论是AD还是VD,均与血管因素有关。慢性脑缺血作为神经系统的一种常见病理状态,是由各种原因导致的长期的脑灌流不足,而导致的脑代谢障碍和功能衰退,是血管性痴呆、Binswanger病、Alzheimer病(AD)等多种疾病发展的一个共同病理过程。脑血流低灌注在痴呆发病机制中的作用日渐成为人们关注热点。为此明确痴呆的发病机制特别是在慢性脑缺血的状态下引发的痴呆的发病机制显得尤为重要。既往对慢性脑缺血导致认知功能的研究多集中在脑代谢、脑血管损害、脑内免疫细胞活动、细胞凋亡、突触结构等方面。 Numb蛋白最初在果蝇周围神经系统的发育中被发现的,因其在果蝇细胞中有决定细胞命运的作用被称为细胞命运决定子,随后在哺乳动物以及人的神经系统也发现了同样的功能,但哺乳动物Numb蛋白的结构和功能较果蝇复杂。在近年的研究中发现Numb作为细胞内吞蛋白在神经发育过程中发挥重要作用。不仅参与调节神经前体细胞的不对称分裂、维持神经祖细胞的自我更新还参与调控神经细胞迁移和神经突生长等。本实验预通过检测慢性脑缺血大鼠认知功能的改变及海马区Numb表达的动态变化初步探讨慢性脑缺血所致的认知功能改变和Numb之间的关系及可能机制。为缺血性脑血管病所引起的认知障碍的研究与治疗提供理论依据。 方法 1.雄性SD大鼠40只,鼠龄大于12个月,随机分为假手术组、缺血8周组、缺血10周组、缺血12周组4组,每组10只。 2.采用永久性双侧颈总动脉结扎制备脑灌注不足模型(2VO模型),假手术组除不接扎双侧颈动脉外余处理与手术组相同。 3.各组在对应时间进行Morris水迷宫试验,检测大鼠的学习和记忆功能。 4.通过免疫组化检测大鼠海马CA1区Numb的表达。 5.通过Western bolt检测大鼠海马的Numb表达 6.数据采集和数据分析。 结果 1.行为学检测结果显示,缺血组大鼠逃避潜伏期较假手术组明显延长,穿越平台次数明显减少,且随着缺血时间的延长,逃避潜伏期进行性延长,穿越平台次数进行性减少,表明2VO术后大鼠学习记忆能力明显受损,且随缺血时间延长进行性加重。 2.免疫组化半定量结果显示,缺血组大鼠海马CA1区Numb表达较假手术组明显减少,并随缺血时间的延长进行性减少。 3.Western bolt结果显示,缺血组大鼠海马CA1区Numb表达较假手术组明显减少,并随缺血时间的延长进行性减少。 结论 2VO术后8周至12周中,随着缺血时间的延长大鼠认知进行性损害、Numb表达进行性下降。Numb表达的进行性下降可能是慢性缺血导致认知功能障碍的重要原因之
[Abstract]:Background and objective with the aging of human society, the incidence of dementia is increasing year by year. Vascular dementia (VD) and Alzheimer's disease (AD) accounted for the majority of dementia in dementia population, and the incidence of VD was slightly higher than that of AD. It brings serious economic and mental burden to the society and the family. In recent years, it is believed that both AD and VD are related to vascular factors. Chronic cerebral ischemia, as a common pathological state of the nervous system, is caused by a variety of causes of chronic cerebral perfusion insufficiency, and resulting in brain metabolic disorders and functional decline. It is a common pathological process in the development of vascular dementia Binswanger disease, Alzheimer's disease, (AD) and other diseases. The role of cerebral blood flow hypoperfusion in the pathogenesis of dementia has become a hot topic. Therefore, it is very important to clarify the pathogenesis of dementia, especially in the condition of chronic cerebral ischemia. Previous studies on cognitive function caused by chronic cerebral ischemia have focused on brain metabolism, cerebral vascular damage, immune cell activity and apoptosis in the brain. Synaptic structure and other aspects. Numb protein was initially found in the development of the peripheral nervous system of Drosophila melanogaster, because of its role in determining cell fate in Drosophila cells, known as cell fate determinant, The same function was found in mammalian and human nervous system, but the structure and function of mammalian Numb protein was more complicated than that of Drosophila. In recent years, Numb has been found to play an important role in neurodevelopment as an endocytosis protein. It is not only involved in regulating asymmetric division of neural precursor cells, maintaining self-renewal of neural progenitor cells, but also in regulating neuronal migration and neuronal growth. The purpose of this study was to explore the relationship between cognitive function and Numb and the possible mechanism of chronic cerebral ischemia by detecting the changes of cognitive function and the dynamic changes of Numb expression in hippocampus of rats with chronic cerebral ischemia. To provide theoretical basis for the study and treatment of cognitive impairment caused by ischemic cerebrovascular disease. Method 1. Forty male Sprague-Dawley rats, aged more than 12 months, were randomly divided into sham-operation group, ischemia 8-week group, ischemic 10-week group and ischemic 12-week group with 10 rats in each group. The model of cerebral perfusion insufficiency (2VO model) was established by permanent ligation of bilateral common carotid artery (2VO model). The remaining treatment in sham-operation group was the same as that in operation group except without ligation of bilateral carotid artery. The Morris water maze test was performed in each group at the corresponding time to detect the learning and memory function of rats. 4. 4. The expression of Numb in rat hippocampal CA1 was detected by immunohistochemistry. 5. 5. The expression of Numb in rat hippocampus was detected by Western bolt. Data acquisition and data analysis. Result 1. The results of behavioral examination showed that the escape latency of ischemic group was significantly longer than that of sham operation group, and the times of crossing the platform were significantly decreased, and with the prolongation of ischemic time, the escape latency was gradually prolonged, and the number of crossing platform was decreased progressively. The results showed that the learning and memory ability of rats after 2VO was obviously impaired, and gradually aggravated with the prolongation of ischemic time. 2. 2. The semi-quantitative immunohistochemical results showed that the expression of Numb in hippocampal CA1 in ischemic group was significantly lower than that in sham-operated group, and gradually decreased with the prolongation of ischemic time. 3.Western bolt results showed that the expression of Numb in hippocampus of ischemic group was significantly lower than that of sham operation group. The expression of Numb in hippocampal CA1 in ischemic group was significantly lower than that in sham operation group, and gradually decreased with the prolongation of ischemic time. Conclusion during the 8-12 weeks after 2VO, the progressive decrease of the expression of numb and the expression of numb may be the important cause of cognitive dysfunction caused by chronic ischemia in rats with progressive cognitive impairment with the prolongation of ischemic time.
【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R749.1
本文编号：2144219
[Abstract]:Background and objective with the aging of human society, the incidence of dementia is increasing year by year. Vascular dementia (VD) and Alzheimer's disease (AD) accounted for the majority of dementia in dementia population, and the incidence of VD was slightly higher than that of AD. It brings serious economic and mental burden to the society and the family. In recent years, it is believed that both AD and VD are related to vascular factors. Chronic cerebral ischemia, as a common pathological state of the nervous system, is caused by a variety of causes of chronic cerebral perfusion insufficiency, and resulting in brain metabolic disorders and functional decline. It is a common pathological process in the development of vascular dementia Binswanger disease, Alzheimer's disease, (AD) and other diseases. The role of cerebral blood flow hypoperfusion in the pathogenesis of dementia has become a hot topic. Therefore, it is very important to clarify the pathogenesis of dementia, especially in the condition of chronic cerebral ischemia. Previous studies on cognitive function caused by chronic cerebral ischemia have focused on brain metabolism, cerebral vascular damage, immune cell activity and apoptosis in the brain. Synaptic structure and other aspects. Numb protein was initially found in the development of the peripheral nervous system of Drosophila melanogaster, because of its role in determining cell fate in Drosophila cells, known as cell fate determinant, The same function was found in mammalian and human nervous system, but the structure and function of mammalian Numb protein was more complicated than that of Drosophila. In recent years, Numb has been found to play an important role in neurodevelopment as an endocytosis protein. It is not only involved in regulating asymmetric division of neural precursor cells, maintaining self-renewal of neural progenitor cells, but also in regulating neuronal migration and neuronal growth. The purpose of this study was to explore the relationship between cognitive function and Numb and the possible mechanism of chronic cerebral ischemia by detecting the changes of cognitive function and the dynamic changes of Numb expression in hippocampus of rats with chronic cerebral ischemia. To provide theoretical basis for the study and treatment of cognitive impairment caused by ischemic cerebrovascular disease. Method 1. Forty male Sprague-Dawley rats, aged more than 12 months, were randomly divided into sham-operation group, ischemia 8-week group, ischemic 10-week group and ischemic 12-week group with 10 rats in each group. The model of cerebral perfusion insufficiency (2VO model) was established by permanent ligation of bilateral common carotid artery (2VO model). The remaining treatment in sham-operation group was the same as that in operation group except without ligation of bilateral carotid artery. The Morris water maze test was performed in each group at the corresponding time to detect the learning and memory function of rats. 4. 4. The expression of Numb in rat hippocampal CA1 was detected by immunohistochemistry. 5. 5. The expression of Numb in rat hippocampus was detected by Western bolt. Data acquisition and data analysis. Result 1. The results of behavioral examination showed that the escape latency of ischemic group was significantly longer than that of sham operation group, and the times of crossing the platform were significantly decreased, and with the prolongation of ischemic time, the escape latency was gradually prolonged, and the number of crossing platform was decreased progressively. The results showed that the learning and memory ability of rats after 2VO was obviously impaired, and gradually aggravated with the prolongation of ischemic time. 2. 2. The semi-quantitative immunohistochemical results showed that the expression of Numb in hippocampal CA1 in ischemic group was significantly lower than that in sham-operated group, and gradually decreased with the prolongation of ischemic time. 3.Western bolt results showed that the expression of Numb in hippocampus of ischemic group was significantly lower than that of sham operation group. The expression of Numb in hippocampal CA1 in ischemic group was significantly lower than that in sham operation group, and gradually decreased with the prolongation of ischemic time. Conclusion during the 8-12 weeks after 2VO, the progressive decrease of the expression of numb and the expression of numb may be the important cause of cognitive dysfunction caused by chronic ischemia in rats with progressive cognitive impairment with the prolongation of ischemic time.
【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R749.1
【参考文献】
相关硕士学位论文 前1条
1 刘秀丽;基于GFP的FRET监测树突棘钙动力学[D];华中科技大学;2005年
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