本文選題：微透析 + 乙酰膽堿�。� 參考：《上海交通大學(xué)》2014年碩士論文

【摘要】：在動(dòng)物體內(nèi)存在有種類繁多，功能各異的神經(jīng)遞質(zhì)。中樞神經(jīng)遞質(zhì)中的乙酰膽堿（Acetylcholine）伽馬氨基丁酸（GABA），是非常重要的生物活性成分，在調(diào)節(jié)生物體的功能上起著非常重要的作用，尤其是在生物體的對(duì)事物的認(rèn)知、記憶等諸多方面。很多證據(jù)表明，乙酰膽堿、伽馬氨基丁酸異常可能引起多種疾病，例如由于缺少引起的老年癡呆癥等。鑒于乙酰膽堿、伽馬氨基丁酸及其各自的代謝產(chǎn)物具有重要的生理學(xué)作用，，故其生理濃度變化的測(cè)量可作為疾病進(jìn)展或藥物治療的評(píng)估指標(biāo)，這一功能使得這類物質(zhì)成為藥物研發(fā)中非常重要的生化標(biāo)記物。但是在利用活體動(dòng)物直接觀測(cè)乙酰膽堿、伽馬氨基丁酸的變化較為困難，為了解決這一難題本試驗(yàn)利用微透析技術(shù)，口服給予多奈哌齊觀測(cè)其對(duì)大鼠紋狀體區(qū)神經(jīng)遞質(zhì)的影響。通過(guò)腦立體定位技術(shù)與腦微透析儀取樣技術(shù)對(duì)清醒狀態(tài)下自由活動(dòng)的大鼠取樣采用離線超高效液相色譜串聯(lián)質(zhì)譜法檢測(cè)取自大腦紋狀體的透析液內(nèi)乙酰膽堿、伽馬氨基丁酸的濃度，并實(shí)現(xiàn)了同時(shí)檢測(cè)兩種化學(xué)遞質(zhì)的定量技術(shù)。在新的藥物研發(fā)過(guò)程中，來(lái)自生物樣本的疾病體采集物中的生化標(biāo)記物常被用作診斷和藥物療效評(píng)估的有效方式。研究多奈哌齊對(duì)大鼠大腦紋狀體內(nèi)神經(jīng)遞質(zhì)乙酰膽堿伽馬氨基丁酸的變化規(guī)律，為微透析技術(shù)在醫(yī)藥研發(fā)中的工業(yè)化應(yīng)用，提供具體可靠的技術(shù)手段和參考，以便快速提高研發(fā)效率和降低研發(fā)成本。 本次實(shí)驗(yàn)建立了中樞神經(jīng)遞質(zhì)中的乙酰膽堿、伽馬氨基丁酸（超高效液相色譜串聯(lián)質(zhì)譜的測(cè)定方法。結(jié)果：線性關(guān)系、方法精密度好、準(zhǔn)確度高，符合分析要求。采用增量法和減量法考察待測(cè)濃度對(duì)探針回收率的影響。結(jié)果：微透析探針回收率基本不受濃度影響，且體內(nèi)回收率穩(wěn)定性良好。結(jié)論:應(yīng)用UPLC-MS/MS技術(shù)建立使用膽堿酯酶抑制劑測(cè)定大鼠腦微透析液中乙酰膽堿、伽馬氨基丁酸的方法迅速、靈敏、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.
【學(xué)位授予單位】：上海交通大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R965

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