广东凉茶作用机制的代谢组学研究

发布时间：2018-04-30 19:50

本文选题：广东凉茶颗粒 + 代谢组学　；参考：《华南理工大学》2012年博士论文

【摘要】：广东凉茶颗粒是多味中草药的水提取物，组分复杂。据传统中医药理论认为，它具有清热解毒等功能，是凉茶的典型代表。随着人们生活水平的提高，以中草药为配方的广东凉茶受到极大关注。广东凉茶作用温和、具有多靶点作用特点，但因其组分复杂而难以按照常规方法对凉茶进行科学评价，因而，应该将广东凉茶视为一个化学物质系统，利用现代代谢组学方法来阐述这个物质系统对于特定生物机体所产生的整体影响，为建立具有良好科学性的凉茶作用评价系统奠定基础。代谢组学是借助高通量、高灵敏度的分析技术和各种信息处理技术研究各种生物样本如体液、细胞及组织中的小分子代谢物质整体对各种刺激的变化，解析各种代谢途径的变化，进而解析机体整体的生理状态，反映生物机体对所有扰动因素进行应答的终端信息，具有整体性、系统性、动态性的特点，故，，代谢组学已广泛应用于疾病、药物、营养、植物等方面的研究。由于代谢组学研究的是外源性物质对生物体代谢物组产生的整体效应，其思想具有与中医药理论相一致的观点，利用代谢组学来研究广东凉茶这类中草药作用机制，将有助于全面了解广东凉茶的作用机理。本研究选择以大鼠作为研究对象，探讨广东凉茶在其机体内所产生的代谢组学规律。主要研究内容如下：一、基于NMR的代谢组学研究方法分析不同饮用时间的大鼠血清样本，发现大鼠的代谢组学响应不一样，表明大鼠对广东凉茶的响应具有动态性特点。对于10d饮用广东凉茶的分析显现，凉茶组大鼠和对照组大鼠没有明显区别，这说明大鼠机体对广东凉茶的响应不明显，也可能是大鼠体内环境稳定所致。但是，20d的凉茶组大鼠则显现出与对照组大鼠明显的差异，大鼠饮用广东凉茶后，血糖水平下降，糖异生水平增加，能量代谢由糖代谢途径转入脂肪代谢。经过30d连续饮用广东凉茶后，凉茶组大鼠和对照组大鼠仍然明显分离，血糖血脂水平降低，但此时血清中丙酮酸水平下降，表明，广东凉茶有可能导致其代谢功能低下。二、利用GC-MS分析技术和PCA、OPLS-DA等多元数据处理方法对为期38d的凉茶饮用阶段里的大鼠尿液代谢物进行分析，发现，反复饮用广东凉茶能够引起体内代谢物的积累效应和众多代谢途径的变化，比如能量代谢、氨基酸代谢等，但是，这些代谢途径出现的变化趋势在整个响应过程中并不一致。因此，对各个取样点的凉茶组大鼠和对照组大鼠尿液代谢物组进行OPLS-DA分析，确定：凉茶组大鼠中前期的主要变化是以抗氧化水平提高、体内过氧化水平降低为主要特征；随着连续饮用时间的延长，这种效应慢慢消失，反映了大鼠机体的自适应过程；但进一步延长饮用时间，大鼠的代谢组响应呈现另外一个特征，即能量流的降低，并出现肠道菌群的紊乱。如果说前期出现的代谢组变化是系统内的平衡态的扰动，那末，在后期出现的能量流降低以及肠道微生物菌群的紊乱，可能意味着系统内的代谢物组偏离了平衡态，这种偏离平衡态的扰动表明，长期连续饮用广东凉茶对大鼠机体可能导致不利影响。三、为进一步验证广东凉茶这个复杂体系对大鼠机体的作用机制，利用拘束应激来模拟“上火”症状，采用血清和尿液代谢组学方法，描述拘束造成的“上火”模型大鼠体内的代谢通路变化规律和广东凉茶对拘束应激造成大鼠代谢紊乱的恢复作用，表明，拘束应激大鼠给予凉茶后，多数受到影响的代谢途径都得到恢复，其中包括能量代谢、肠道微生物组成的恢复以及各种涉及机体免疫力和应激能力的氨基酸代谢水平的升高。除此之外，应激大鼠在给予凉茶后，机体抗氧化能力增强，胆固醇水平降低。尽管如此，引起拘束应激大鼠中代谢显著变化的喹啉酸并没有完全恢复，表明拘束应激造成神经系统功能的障碍并未能通过广东凉茶得到完全改善。
[Abstract]:Guangdong herbal tea granule is a water extract of Chinese herbal medicine with complex components. According to the traditional Chinese medicine theory, it has the function of clearing heat and detoxification. It is a typical representative of cold tea. With the improvement of people's living standard, Guangdong herbal tea with Chinese herbal medicine as the formula is of great concern. The effect of cold tea in Guangdong is mild, but it has the characteristic of multi target. Because of its complex composition, it is difficult to scientifically evaluate the herbal tea in accordance with conventional methods. Therefore, Guangdong herbal tea should be regarded as a chemical substance system, and the modern metabonomics method is used to explain the overall effect of the material system on the specific organism, and to establish a good and scientific evaluation system for the effect of cold tea. Set the foundation.
Metabonomics is the use of high throughput, high sensitivity analysis techniques and various information processing techniques to study the changes in various stimuli in various biological samples, such as body fluids, cells and tissues, to analyze the changes in various metabolic pathways, and then to analyze the physiological state of the body, reflecting the disturbance of the organism to all disturbances. The terminal information that responds to factors has the characteristics of holistic, systematic, and dynamic. Therefore, metabonomics has been widely used in the research of diseases, drugs, nutrition, plants and so on. Because the metabolomics research is the overall effect of exogenous substances on organism metabolites, the idea has a consistent view with the theory of traditional Chinese medicine. Point of view, using metabonomics to study the action mechanism of herbal medicine such as Guangdong herbal tea will help us to understand the mechanism of Guangdong herbal tea in a comprehensive way.
In this study, rats were selected as the research objects to explore the metabolomics law of Guangdong herbal tea in its body.
First, based on the NMR metabonomics method, the serum samples of rats with different drinking time were analyzed, and the metabolic response of rats was different, which showed that the response of rats to Guangdong cold tea was dynamic. The analysis of 10d drinking cold tea showed that there was no obvious difference between the cold tea group rats and the control rats, which indicated that the rats were different from those in the control group. The response of the body to Guangdong cold tea was not obvious, but it may also be the result of the stable environment in the rats. However, the cold tea group of 20d showed obvious difference with the control group. After drinking Guangdong cold tea, the blood sugar level decreased, the sugar ISO level increased, the energy metabolism was transferred from sugar metabolism to fat metabolism. After 30d, the rats were drinking widely. After the eastern cold tea, the rats in the cold tea group and the control group were still clearly separated and the blood glucose and blood lipid level decreased, but the level of the pyruvate in the serum decreased, indicating that the Guangdong herbal tea may lead to its low metabolic function.
Two, GC-MS analysis technology and PCA, OPLS-DA and other multivariate data processing methods were used to analyze the urine metabolites of rats in the drinking stage of cold tea for a period of 38d. It was found that repeated drinking of Guangdong cold tea could cause the accumulation effect of metabolites in the body and the changes of many metabolic pathways, such as energy metabolism, amino acid metabolism, etc. The variation trend of the metabolic pathway was not consistent throughout the response process. Therefore, the OPLS-DA analysis of the urine metabolite group in the cold tea group rats and the control group was made to determine that the main changes in the early stage of the cold tea group were that the antioxidant level was increased and the level of the peroxidation in the body decreased as the main characteristic; When the drinking time was prolonged, this effect slowly disappeared, reflecting the self-adaptive process of the rat body; but further prolonging the drinking time, the response of the metabolic group in the rat showed another characteristic, that is, the decrease of the energy flow and the disorder of the intestinal flora. At the end, the decrease of energy flow in the late stage and the disturbance of intestinal microflora may mean that the metabolites in the system deviate from the equilibrium state, and this deviation from the equilibrium state indicates that the long-term continuous drinking of Guangdong cold tea may have adverse effects on the rat body.
Three, in order to further verify the mechanism of the complex system of Guangdong herbal tea on the rat body, use restraint stress to simulate the symptoms of "upper fire", and use the methods of serum and urine metabolomics to describe the changes of metabolic pathways in the "upper fire" model rats and the metabolic turbulence caused by restraint stress in Guangdong cold tea. The effect of the disorder showed that most of the metabolic pathways were restored after the restraint stress rats were given cold tea, including energy metabolism, the recovery of the composition of intestinal microbes and the increase of various amino acid metabolism levels that involved the immunity and stress of the body. In addition, the stress rats were resistant to cold tea. In spite of this, quinoline acid, which causes significant metabolic changes in the restraint stress rats, has not been fully recovered, suggesting that the constraint of restraint stress on nervous system function has not been completely improved through Guangdong cold tea.

【学位授予单位】：华南理工大学
【学位级别】：博士
【学位授予年份】：2012
【分类号】：R151

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