【作者】 芦林林；

【导师】 武滨；

【作者基本信息】 山西大学 ， 药物化学， 2012， 硕士

【摘要】 选题依据：抑郁症是一大类心理障碍,一般统称为“情绪障碍”。随着人们的生活压力越来越大,抑郁症的发病几率也越来越高。世界卫生组织已将其与癌症、艾滋病共同列为二十一世纪三大黑死病。出自于宋代《太平惠民和剂局方》的逍遥散在治疗抑郁症方面已经得到了现代临床效果的验证。但是,对于逍遥散抗抑郁的有效部位仍不明确。代谢组学作为一门新兴科学,在医学领域已经得到了广泛的应用,它采用核磁共振(NMR)、气相色谱-质谱联用(GC-MS)等现代仪器分析手段,分析生物样本如血液及尿液组织中内源性代谢物的变化,系统地分析机体生理病理状态的变化,而这种系统性与中医学的整体观相吻合。目的：验证逍遥散抗抑郁有效部位及研究有效部位抗抑郁的作用机理。方法：通过一系列的分离流程,得到逍遥散的石油醚部位(XY-A)、过大孔吸附树脂得30%、60%、95%乙醇洗脱部位(XY-B、 XY-C和XY-D)、药渣水提部位(XY-E)。通过复制慢性不可抑制轻度应激(CUMS)模型,选用雄性成年SD大鼠分别设模型组(MS)、空白组(NS)、石油醚组(XY-A)、30%乙醇洗脱组(XY-B)、60%乙醇洗脱组(XY-C)、95%乙醇洗脱组(XY-D)、药渣水提组(XY-E)、逍遥散复方组(XYS)8个组,给药组每日造模前给药。造模第21天采用代谢笼法收集尿液,并于造模结束后第22天收集血液,采用NMR、 GC-MS分析技术对所得生物样本进行测定,并采用SIMCA-P软件进行多元统计分析,对逍遥散抗抑郁有效部位及其抗抑郁作用机理进行分析。结果：(1)代谢组学分析结果：两种分析技术所得数据经多元统计分析均显示正常对照组和模型组能明显分开,表明造模成功,CUMS刺激引起了模型组代谢轮廓的变化。同时结合相应的载荷图和独立样本t检验,找到了引起差异的潜在生物标志物。正常对照组、模型组和逍遥散各部位组多元统计分析结果显示了逍遥散不同部位组对CUMS引起代谢物变化的不同的干预作用,其中逍遥散石油醚部位组(XY-A)与其他各部位组相比,离正常对照组最近,离模型组最远,表明其有明显的抗抑郁效果,效果最佳,且优于逍遥散复方组。逍遥散不同部位组可不同程度地调节潜在生物标志物趋向于正常对照组,而XY-A部位组可调节大部分的潜在生物标志物的水平趋向于正常对照组。(2)逍遥散有效部位抗抑郁作用机理研究：逍遥散石油醚部位组减弱或取消了外界刺激因素对机体的作用,使CUMS大鼠的代谢轮廓向正常对照组回归,其作用机理可能与琥珀酸、乳酸和甘氨酸等潜在生物标志物有关,从而调节了能量代谢、神经系统等生化代谢途径。结论：本文成功地复制了慢性不可预知应激模型并通过对采集的生物样本(尿液、血液)的代谢组学分析,确定了逍遥散抗抑郁的有效部位和有效部位的抗抑郁作用机理。从逍遥散分离得到的五个不同极性部位,石油醚部位(XY-A)的抗抑郁效果最好,且优于逍遥散复方,这将为逍遥散的药效物质基础研究提供新的思路,为抗抑郁新药的开发和研究提供一定的基础。更多还原

【Abstract】 RATIONALES:Depression is a serious mental illness, which is called as the "emotional disorder". With the increasing pressure of people’s lives, the incidence of depression is the chance of getting higher and higher. It has been thank as the three black plague of the twenty-first century along cancer and AIDS by World Health Organization. Xiaoyaosan is from "Taiping huimin heji jufang" of Song Dynasty in the treatment of depression has been the validation of the clinical effects. However, the antidepressant effective fractions of the Xiaoyaosan is not clear. As an emerging science, metabonomics has been widely used in the medical field. The analytical methods like nuclear magnetic resonance (NMR), gas chromatography-mass spectrometry (GC-MS) and so on were used to analysis the change of endogenous metabolites from biological samples such as blood and urine and to analysis the body’s physiological and pathological changes. The systemic feature is consistent with the principle of TCM.OBJECTIVE:To explore the active fraction of Xiaoyaosan in producing antidepressant effect and to achieve the antidepressant mechanisms of the active fraction of Xiaoyaosan.METHODS:Xiaoyaosan through a series of separation processes, petroleum ether fraction (XY-A),30%ethanol fraction (XY-B),60%ethanol fraction (XY-C),95%ethanol fraction (XY-D) and water fraction (XY-E) were obtained. Through copying the chronic unpredictable stress model, the healthy adult male Sprague-Dawley (SD) rats were divided into seven groups:control group, chronic unpredictable stress model group, petroleum ether fraction group,30%ethanol fraction group,60%ethanol fraction group,95%ethanol fraction group and water fraction group. Urine samples were collected in21days. Plasma samples were collected on22day. The NMR and GC-MS analysis techniques were applied to determine the biological samples. The SIMCA-P software for the multivariate analysis was used to analysis the active fraction of Xiaoyaosan in producing antidepressant effect and the antidepressant mechanisms of the active fraction of Xiaoyaosan.RESULTS:(1) The results of metabolic studying:The data obtained from the two analysis techniques through multivariate statistical analysis showed the control group and model group can be clearly separated, which showed the model was successful. With the corresponding loading plot and independent sample T test, the potential biomarkers were found. The multivariate statistical analysis results for control group, model group and different fractions of Xiaoyaosan show that the different intervention effect of different fractions of Xiaoyaosan to the change of metabolites. Compared with the other fractions of Xiaoyaosan, the petroleum ether fraction of Xiaoyaosan was much closer to the control group and was the farthest away from the model group, indicating its significant antidepressant effect. The different fractions of Xiaoyaosan can regulate potential biomarkers tend to the normal control group, while the petroleum ether fraction of Xiaoyaosan can regulate most potential biomarker levels tend to control group.(2) Antidepressant mechanism research of active fraction of Xiaoyaosan:the petroleum ether fraction of Xiaoyaosan abate or cancel the stimulus factors on the function of organism, so that the metabolic spectrum of CUMS rats tend to the control group. Its mechanism of action may be related to potential biomarkers likes succinic acid, lactic acid and glycine. They can regulate the aspect of energy metabolism, nerve system and so on.CONCLUSION:The article copied the chronic unpredictable stress model successfully and analysis the collection of biological samples (urine, plasma) through metabolomics. The results identified the active fraction of Xiaoyaosan in producing antidepression and antidepressant mechanism of active fraction of Xiaoyaosan. The antidepressant effect of petroleum ether fraction is best in five different polar fractions of Xiaoyaosan, This will provide the new ideas for the study of material basis for the efficacy of Xiaoyaosan and provide the basis for the new drug research.更多还原