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题名	猕猴脑内甲醇代谢的研究
作者	郑娜
学位类别	硕士
答辩日期	2015-05
授予单位	中国科学院研究生院
授予地点	北京
导师	胡新天
关键词	猕猴 甲醇 甲醛 脑脊液
中文摘要	阿尔茨海默病(Alzheimer’s disease, AD)是一种神经退行性脑疾病。我们实验室采用口服甲醇的方法可建立新型的猕猴AD模型，在表现出了以淀粉样沉淀和tau蛋白过度磷酸化的典型AD病理特征的同时，动物还出现了记忆损伤这一特征性临床症状。这是国内外首个具有所有AD重要特征的猕猴动物模型。因为口服甲醇及代谢产物甲醛对外周组织造成损伤，机理不容易阐明，所以我们进一步通过脑内微量注射甲醇的方法，也成功建立了十分相似的猕猴AD模型。为了阐明这一新型模型的成模机理，探索甲醇进入脑内的代谢情况十分关键。另外，我们成功建立AD模型提示：甲醇对大脑有明显毒性。目前甲醇对神经系统的毒性研究主要集中在对视网膜的急性损伤，但是，甲醇对大脑本身的毒性目前还没有研究。为了阐明新型AD模型的成模机理以及甲醇对大脑产生毒性的原理，根据赫荣乔老师提出的甲醛可能诱发AD的理论，我们提出如下假说：甲醇通过脑内相关酶代谢为甲醛, 造成脑损伤，进而发展为AD。但是，目前还没有关于甲醇在非人灵长类脑内代谢过程的具体研究报道。 本实验通过向猕猴侧脑室微量注射甲醇，采用甲醛试剂盒检测脑脊液中甲醛浓度随时间的变化关系来研究甲醇在脑内的代谢过程。实验中一共使用了6只雄性猕猴，3只短期给药，3只长期连续给药8个月以上。短期给药的结果表明：在给药3 h后，脑内甲醛的含量逐渐升高，到18 h达到峰值，然后逐渐下降，最后恢复至正常的生理平衡水平。长期给药的结果表明：在给药第2个月，脑内甲醛含量没有增高，在给药第4、6、8个月，甲醛的浓度表现出逐渐升高的趋势。这是国内外首次对非人灵长类脑内甲醇代谢参数进行系统的测量。另外，为了确证试剂盒检测脑内甲醛浓度这一方法的可靠性，我们还采用高效液相色谱法对同一样品进行了测试，通过比较二者的结果证明我们的检测方法是可靠的。 总之，本文表明：不管是短期给药还是长期给药，甲醇进入脑内能够被代谢生成甲醛。该结论为脑内给甲醇建立猕猴AD模型提供有效的基础数据，并可对将来甲醛建立AD模型起指导作用。 在上面工作的基础上，下一步就应该研究脑内甲醇代谢成为甲醛的过程与猕猴学习记忆损伤过程之间的联系，因此，我训练了猕猴完成工作记忆任务。目前，训练的2只猕猴已完成1000个测试，确定了测试任务初始值，下一步训练将改变延缓时间，完成整个训练过程，为进一步研究脑内甲醇代谢过程与猕猴学习记忆损伤过程的关系提供了基础。
英文摘要	Alzheimer’s disease(AD) is a progressive neurodegenerative disease. Methanol feeding monkey model was established in previous study in our lab, the monkeys have AD’s most important characteristics, including tau hyperphosphorylation, amyloid deposits as well as the working memory impairment. This model is one of the success models which have most of AD symptoms. In the present study, another (a modified) similar methanol treat AD model was induced by lateral ventricles injection, which could facilitate exploration the metabolic procession of methanol in brain tissue and acquired the key parameters of it, and most important, with no damage to peripheral tissue but the brain, furthermore, the mechanism of this model is much easier to elaborate. Now the key point to explain the model mechanism is to examine the methanol metabolites. In another hand, our success AD model showed that methanol is toxicity to brain, at present, methanol toxicity studies mostly focused on the optical system, but, only a few on brain. This experiment is designed to elaborate the mechanism of our methanol injection monkey model and the principle of methanol toxicity to the brain. According to Professor He Rongqiao’s theory “formaldehyde may induce AD-related characteristics”, we have this hypothesis: methanol can be metabolized to formaldehyde in the brain, and then lead to the brain damage which may cause AD gradually. And there is no study on methanol metabolism in non-human primate’s brain till now. The time-dependent variations of the formaldehyde concentration in cerebrospinal fluid were detected by formaldehyde assay kit. We used six male macaques in the experiment, three were treated with methanol short-term, and the other three were treated with methanol more than eight months. Consequently, the concentration of formaldehyde started to increase gradually after 3h administration of methanol and reached to a peak about 18h later, then began to decrease, and recovered to the physiological equilibrium in normal level at last. The long-term results showed that the concentration of formaldehyde was not increased in the first two months, but exhibited an increasing trend after 4, 6, 8 months administration of methanol. High performance liquid chromatography (HPLC) was used to detect on the same sample to verify the reliability of result. In summary, the formaldehyde concentration raised in short-term and long-term observation after treated with methanol. The result suggested that methanol can generate formaldehyde in the brain. This result can provide the preliminary data base for the building of the AD model in monkeys using methanol and guide for formaldehyde leading to AD model in monkeys later. Based on the above work, I participated in macaques working memory task training. At present, two monkeys have finished 1000 trails and determined the initial value of test task, and the next step will be trained on variable delay response task. This provides an important basis for further study of relationship between methanol metabolism in brain and memory impairment in rhesus monkey.
语种	中文
内容类型	学位论文
源URL	[http://159.226.149.26:8080/handle/152453/10087]
专题	昆明动物研究所_神经系统编码
作者单位	中国科学院昆明动物研究所
推荐引用方式 GB/T 7714	郑娜. 猕猴脑内甲醇代谢的研究[D]. 北京. 中国科学院研究生院. 2015.

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