文拉法辛作用于星形胶质细胞基于~1HNMR的代谢组学研究

发布时间：2018-04-29 11:06

本文选题：文拉法辛 + 代谢组学　；参考：《重庆医科大学》2017年硕士论文

【摘要】：研究背景抑郁症(Major depressive disorder,MDD)是一种常见的可致残的严重精神性疾病,抑郁症与其他一些慢性疾病相比(像糖尿病,高血压,关节炎等疾病)具有高患病率并对全球人类的健康状况有着巨大的威胁。然而,目前抑郁症的发病机制以及相关诊断相对主要根据患者的临床表现,而确切诊断则存在很大困难,文拉法辛是5-羟色胺和去甲肾上腺素再摄取抑制剂(serotonin-norepinephrine reuptake inhibitors SNRIs),多巴胺再摄取的抑制作用较弱。但是,目前关于文拉法辛抗抑郁治疗的具体细胞与分子层面的作用机制并不是十分明确。代谢组学被认为是一种有效且新颖的方法被应用于对文拉法辛药物机理的研究中。IPA(Ingenuity Pathways Analysis)可在生命科学研究的核心中建模并分析理解复杂生物和化学系统,帮助发掘数据的价值。星形胶质细胞(Astrocytes,Ast)是处于中枢神经系统中(Central nervous system,CNS)的一种的细胞。开展文拉法辛在星形胶质细胞上的代谢组学研究将给抑郁症的治疗带来新的思路,提供更多的证据与科研线索。目的文拉法辛作为抗抑郁的新药物,据越来越多的研究表明,星形胶质细胞和文拉法辛的抗抑郁作用有着紧密关联。我们通过将文拉法辛作用于原代星形胶质细胞并进行代谢组学与生物信息学分析,深入探讨文拉法辛对星形胶质细胞的作用机制或可为抑郁症的治疗提供一些新的研究思路和研究证据。方法1.星形胶质细胞原代培养2.用CCK-8进行细胞增殖实验检测来选择文拉法辛处理的星形胶质细胞的最优浓度(最优浓度为10μM)。3.将文拉法辛处理组与正常组对照组进行~1HNMR代谢组学分析,再采用多元统计分析方法,如最小二乘法分析(Partial least squares-discriminate analysis,PLS-DA),正交偏最小二乘法判别分析(Orthogonal partial least-squares discriminant analysis,OPLS-DA)等,最后进行IPA生物消息学分析。结果1.构建高效成熟稳定的星形胶质细胞的原代培养方法。2.在进行多元统计学分析后显示,加药组与正常对照组中共有31种代谢物指标有着明显的差异,这些代谢物主要参与能量代谢,脂质代谢,氨基酸代谢。3.通过生物信息学分析文拉法辛加药组与氟西汀加药组的差异代谢物,可得出两组药物的代谢图谱的异同点。结论基于~1HNMR代谢组学方法探讨文拉法辛对星形胶质细胞作用后所引起的一系列内源性代谢通路变化与其中主要的代谢差异物,并通过生物信息学分析结果表明共31种代谢物与能量代谢,氨基酸代谢,脂质代谢通路有关。为文拉法辛对抑郁症的治疗机制提供更深的理解并提供更多的实验依据。
[Abstract]:Background Major depressive disorder (MDD) is a common, disabling, severe psychiatric disorder compared with other chronic diseases (such as diabetes, hypertension, hypertension). Arthritis and other diseases have a high prevalence and pose a great threat to human health worldwide. However, at present, the pathogenesis and related diagnosis of depression are mainly based on the clinical manifestations of the patients, but the exact diagnosis is very difficult. Venlafaxine is a serotonin-norepinephrine reuptake inhibitors SNR Isa inhibitor of serotonin-norepinephrine and norepinephrine, and the inhibitory effect of dopamine reuptake is weak. However, the cellular and molecular mechanisms of venlafaxine antidepressant therapy are not clear. Metabonomics is considered to be an effective and novel method for studying the mechanism of venlafaxine drugs. IPA-Ingenuity Pathways Analysis can model and analyze complex biological and chemical systems at the core of life sciences research, and help to discover the value of data. Astrocytes steroid is a cell in the central nervous system (CNS). The research on metabonomics of venlafaxine on astrocytes will bring new ideas to the treatment of depression and provide more evidence and scientific clues. Objective venlafaxine is a new antidepressant. More and more studies have shown that astrocytes and venlafaxine are closely related to antidepressant effects. We used venlafaxine to act on primary astrocytes and to analyze metabonomics and bioinformatics. Further study on the mechanism of venlafaxine on astrocytes may provide some new ideas and evidences for the treatment of depression. Method 1. Astrocytes were cultured in primary culture. The optimal concentration of the astrocytes treated with venlafaxine was determined by CCK-8 cell proliferation assay (the optimal concentration was 10 渭 MU. 3). The control group and the control group were treated with venlafaxine and analyzed by 1H NMR metabolomics. The methods of multivariate statistical analysis, such as partial least squares-discriminate analysis PLS-DAA, orthogonal partial least-squares discriminant analysis, OPLS-DAA, etc., were used. Finally, IPA bioinformatics analysis was carried out. Result 1. The primary culture method of constructing high efficient mature and stable astrocytes. 2. After multivariate statistical analysis, there were significant differences in 31 metabolites between the admixture group and the normal control group. These metabolites were mainly involved in energy metabolism, lipid metabolism and amino acid metabolism. By bioinformatics analysis of the different metabolites between venlafaxine group and fluoxetine group, the similarities and differences of metabolic maps between the two groups were obtained. Conclusion A series of endogenous metabolic pathways induced by venlafaxine on astrocytes and the main metabolic differences were investigated based on 1H NMR metabolomics. The results of bioinformatics analysis showed that 31 metabolites were related to energy metabolism, amino acid metabolism and lipid metabolism pathway. To provide a deeper understanding of the treatment mechanism of depression and provide more experimental evidence for venlafaxine.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R749.4

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