多奈哌齐对大鼠纹状体内乙酰胆碱和Y-氨基丁酸的影响

发布时间：2018-04-10 13:10

本文选题：微透析 + 乙酰胆碱　；参考：《上海交通大学》2014年硕士论文

【摘要】：在动物体内存在有种类繁多，功能各异的神经递质。中枢神经递质中的乙酰胆碱（Acetylcholine）伽马氨基丁酸（GABA），是非常重要的生物活性成分，在调节生物体的功能上起着非常重要的作用，尤其是在生物体的对事物的认知、记忆等诸多方面。很多证据表明，乙酰胆碱、伽马氨基丁酸异常可能引起多种疾病，例如由于缺少引起的老年痴呆症等。鉴于乙酰胆碱、伽马氨基丁酸及其各自的代谢产物具有重要的生理学作用，，故其生理浓度变化的测量可作为疾病进展或药物治疗的评估指标，这一功能使得这类物质成为药物研发中非常重要的生化标记物。但是在利用活体动物直接观测乙酰胆碱、伽马氨基丁酸的变化较为困难，为了解决这一难题本试验利用微透析技术，口服给予多奈哌齐观测其对大鼠纹状体区神经递质的影响。通过脑立体定位技术与脑微透析仪取样技术对清醒状态下自由活动的大鼠取样采用离线超高效液相色谱串联质谱法检测取自大脑纹状体的透析液内乙酰胆碱、伽马氨基丁酸的浓度，并实现了同时检测两种化学递质的定量技术。在新的药物研发过程中，来自生物样本的疾病体采集物中的生化标记物常被用作诊断和药物疗效评估的有效方式。研究多奈哌齐对大鼠大脑纹状体内神经递质乙酰胆碱伽马氨基丁酸的变化规律，为微透析技术在医药研发中的工业化应用，提供具体可靠的技术手段和参考，以便快速提高研发效率和降低研发成本。本次实验建立了中枢神经递质中的乙酰胆碱、伽马氨基丁酸（超高效液相色谱串联质谱的测定方法。结果：线性关系、方法精密度好、准确度高，符合分析要求。采用增量法和减量法考察待测浓度对探针回收率的影响。结果：微透析探针回收率基本不受浓度影响，且体内回收率稳定性良好。结论:应用UPLC-MS/MS技术建立使用胆碱酯酶抑制剂测定大鼠脑微透析液中乙酰胆碱、伽马氨基丁酸的方法迅速、灵敏、可靠。
[Abstract]:There are many kinds of neurotransmitters in animals with different functions.The acetylcholine acetylcholine gamma-aminobutyric acid (GABA) in central neurotransmitters is a very important bioactive component, which plays a very important role in regulating the functions of organisms, especially in the cognition and memory of things.There is much evidence that abnormal acetylcholine and gamma-aminobutyric acid may lead to many diseases, such as Alzheimer's disease due to lack.Since acetylcholine, gamma-aminobutyric acid and their respective metabolites play an important physiological role, the measurement of their physiological concentrations can be used as an indicator of disease progression or drug therapy,This function makes these substances a very important biochemical marker in drug development.However, it is difficult to observe the changes of acetylcholine and gamma-aminobutyric acid in living animals. In order to solve this problem, the effect of oral administration of Donepezil on the neurotransmitters in the striatum of rats was observed by microdialysis.Acetylcholine (acetylcholine) in dialysate from the striatum of brain was detected by off-line ultra-high performance liquid chromatography-tandem mass spectrometry (HPLC-MS).The concentration of gamma-aminobutyric acid and the quantitative technique for simultaneous detection of two chemical transmitters were realized.In the process of new drug development, biochemical markers in disease samples from biological samples are often used as an effective way to diagnose and evaluate the efficacy of drugs.To study the effect of Donepezil on acetylcholine gamma-aminobutyric acid (GABA), a neurotransmitter in rat brain striatum, and to provide specific and reliable technical means and references for the industrialized application of microdialysis technology in medical research and development.In order to quickly improve R & D efficiency and reduce R & D costs.A method for the determination of acetylcholine and gamma-aminobutyric acid in neurotransmitters was established.Results: linear relationship, good precision, high accuracy, accord with the requirement of analysis.The effect of the measured concentration on the probe recovery was investigated by incremental method and decrement method.Results: the recovery rate of microdialysis probe was not affected by the concentration, and the recovery was stable in vivo.Conclusion: a rapid, sensitive and reliable method for the determination of acetylcholine and gamma-aminobutyric acid in rat brain microdialysis solution using cholinesterase inhibitor was established by UPLC-MS/MS technique.
【学位授予单位】：上海交通大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R965

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