薄荷醇吸嗅改善大鼠空间记忆能力及海马区AchE与GluR1表达的研究

发布时间：2018-03-06 09:23

本文选题：嗅觉通路　切入点：薄荷醇　出处：《安徽医科大学》2012年硕士论文　论文类型：学位论文

【摘要】：研究背景和研究目的老年痴呆症（阿尔茨海默病，Alzheimer’s disease，AD）是临床上常见的精神性疾病。自1906年发现以来，经过一个多世纪的研究，至今仍然有大量问题有待解决。目前治疗AD的药物，如乙酰胆碱酯酶（AchE）抑制剂，只能减轻AD患者的症状，但不能阻止病情的进展。嗅觉系统提供了大脑与外部环境的直接联系，与脑内的许多掌管学习记忆的脑区密切相关，这使得嗅觉通路成为潜在的治疗学习记忆衰退的有效途径。芳香物质如薄荷醇等被认为具有提神醒脑，芳香开窍等作用。本实验就是研究嗅觉通路与学习记忆之间的关系，并通过挥发性的芳香物质薄荷醇对大鼠学习记忆能力的影响，评价其在治疗AD等痴呆症中的作用；为AD的嗅觉通路治疗提供理论依据，开辟AD治疗的新领域；并为芳香疗法提供基础理论支持。第一部分薄荷醇吸嗅对大鼠学习记忆能力的神经行为学观察实验方法将2g的薄荷醇溶于1000ml热水中充分溶解，置于密闭染毒箱中后，放入大鼠作用1h。连续7d，应用Morris水迷宫进行神经行为学检测，通过定位航行试验和空间探索实验观察大鼠空间学习记忆能力的变化。实验结果神经行为学的结果显示，吸入薄荷醇的SD大鼠，定位航行试验和空间探索实验与对照组比较，逃避潜伏期和平台象限滞留时间明显减少（P0.05），说明通过薄荷醇吸嗅后，大鼠的空间学习记忆能力明显提高。第二部分薄荷醇吸嗅大鼠海马区AchE与GluR1免疫组化观察实验方法Morris水迷宫的定位航行试验和空间探索分析表明，吸嗅薄荷醇的SD大鼠的空间学习记忆能力明显提高（P0.05）。为了观察神经行为学变化与神经递质的关系，，通过免疫组织化学的方法，研究薄荷醇吸嗅后对大鼠海马区AchE及GluR1表达水平的变化，探讨其引起大鼠的空间学习记忆能力提高的机制。实验结果脑组织的免疫组化结果显示，吸入薄荷醇的SD大鼠，与对照组相比较，海马区AchE和GluR1的蛋白表达均明显降低（P0.01），积分光密度值和阳性细胞数也明显减少（P0.01）。第三部分薄荷醇吸嗅大鼠海马区AchE与GluR1表达的分子生物学实验观察实验方法为了进一步观察神经递质的变化，通过RT-PCR和ELISA的方法，研究薄荷醇吸嗅后对大鼠海马区AchE及GluR1表达水平的变化，探讨其引起大鼠的空间学习记忆能力提高的机制。实验结果脑组织的RT-PCR结果显示，吸入薄荷醇的SD大鼠，与对照组相比较，海马区AchE和GluR1的mRNA表达均明显降低（P0.01）。ELISA结果表明，吸入薄荷醇的SD大鼠，与对照组相比较，海马区AchE的表达明显降低。结论 1.发现薄荷醇通过嗅觉通路可以明显提高大鼠的学习记忆能力; 2.薄荷醇通过嗅觉通路吸嗅后可以明显降低大鼠海马区AchE和GluR1的表达量； 3.薄荷醇通过嗅觉通路吸嗅后明显提高大鼠的学习记忆能力主要是过降低大鼠海马区AchE的表达量实现的； 4.薄荷醇吸嗅后提高大鼠的学习记忆能力，开辟了治疗AD的新领域。
[Abstract]:Research background and purpose
Senile dementia (Alzheimer's disease, "Alzheimer s disease, AD) is a common clinical mental illness. Since its discovery in 1906, after more than a century of research, there is still a lot of problems to be solved. At present the treatment of AD drugs, such as acetylcholinesterase (AchE) inhibitors, can only alleviate the symptoms of patients with AD. Progress but not prevent the disease. The olfactory system provide a direct connection between the brain and the external environment, closely related to learning and memory in many areas of the brain and the brain, which makes the olfactory pathway as a potential treatment of learning memory decline. Remember the effective way of aromatic substances such as menthol is considered a refreshing, aromatic resuscitation effect etc. this experiment is to study the relationship between. The olfactory pathway of learning and memory, and the effects of volatile aromatic substances of menthol on the ability of learning and memory in rats and its evaluation in the treatment of AD. The role of dementia; providing a theoretical basis for the treatment of AD's olfactory pathway, opening up a new field of AD treatment, and providing basic theoretical support for aromatherapy.
The first part of the neurobehavioral observation of the learning and memory ability of rats by menthol sniffing
Experimental method
The menthol 2G was dissolved in 1000ml hot water and dissolved in a closed poison box. After putting it into a closed poison box, rats were put into 1h. for continuous 7d. The Morris water maze was used to perform neurobehavioral tests, and the spatial learning and memory ability of rats was observed by location navigation test and space exploration experiment.
experimental result
Behavioral results showed that inhalation of menthol in SD rats, positioning navigation test and spatial probe test compared with the control group, the escape latency and the platform quadrant time was significantly reduced (P0.05), illustrated by menthol sniffing, rat spatial learning and memory ability were improved.
Immunohistochemical observation of AchE and GluR1 in the hippocampus of the second part of menthol sniffing rats
The positioning experiment method of Morris water maze navigation test and spatial probe analysis showed that SD rats sniff the menthol in the spatial learning and memory ability was improved significantly (P0.05). To observe the relationship between neural and behavioral changes of neurotransmitters, through the method of immunohistochemistry, research the expression level of menthol sniffing after change on AchE and GluR1 in hippocampus of rats, investigate the mechanism of rat spatial learning and memory ability caused by it.
experimental result
Immunohistochemical results of brain tissue showed that compared with the control group, the expression of AchE and GluR1 protein in hippocampus of SD rats inhaled menthol decreased significantly (P0.01), the integral optical density and the number of positive cells also decreased significantly (P0.01).
The molecular biological experimental observation of the expression of AchE and GluR1 in the hippocampus of the third part of menthol sniffing rats
Experimental method
In order to further observe the changes of neurotransmitters, we studied the changes of AchE and GluR1 expression in hippocampus of rats after menorhion absorption by RT-PCR and ELISA, and explored the mechanism of improving the ability of spatial learning and memory in rats.
experimental result
The RT-PCR results of brain tissue showed that compared with the control group, the mRNA expression of AchE and GluR1 in hippocampus of SD rats inhaled menthol decreased significantly (P0.01).ELISA. The results showed that the expression of AchE in hippocampus of rats inhaled menthol significantly decreased compared with control group.
conclusion
1. it was found that menthol could significantly improve the learning and memory ability of rats through the olfactory pathway.
2. menthol can obviously reduce the expression of AchE and GluR1 in the hippocampus of rats after sniffing through the olfactory pathway.
3. menthol obviously improves the learning and memory ability of rats after sniffing through the olfactory pathway, which is mainly due to the reduction of the expression of AchE in the hippocampus of rats.
4. menthol, after sniffing, improves the learning and memory ability of rats, and opens up a new field for the treatment of AD.

【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R285.5;R749.126

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