本文選題：廣東涼茶顆粒 + 代謝組學(xué)��； 參考：《華南理工大學(xué)》2012年博士論文

【摘要】：廣東涼茶顆粒是多味中草藥的水提取物，組分復(fù)雜。據(jù)傳統(tǒng)中醫(yī)藥理論認(rèn)為，它具有清熱解毒等功能，是涼茶的典型代表。隨著人們生活水平的提高，以中草藥為配方的廣東涼茶受到極大關(guān)注。廣東涼茶作用溫和、具有多靶點(diǎn)作用特點(diǎn)，但因其組分復(fù)雜而難以按照常規(guī)方法對(duì)涼茶進(jìn)行科學(xué)評(píng)價(jià)，因而，應(yīng)該將廣東涼茶視為一個(gè)化學(xué)物質(zhì)系統(tǒng)，利用現(xiàn)代代謝組學(xué)方法來闡述這個(gè)物質(zhì)系統(tǒng)對(duì)于特定生物機(jī)體所產(chǎn)生的整體影響，為建立具有良好科學(xué)性的涼茶作用評(píng)價(jià)系統(tǒng)奠定基礎(chǔ)。 代謝組學(xué)是借助高通量、高靈敏度的分析技術(shù)和各種信息處理技術(shù)研究各種生物樣本如體液、細(xì)胞及組織中的小分子代謝物質(zhì)整體對(duì)各種刺激的變化，解析各種代謝途徑的變化，進(jìn)而解析機(jī)體整體的生理狀態(tài)，反映生物機(jī)體對(duì)所有擾動(dòng)因素進(jìn)行應(yīng)答的終端信息，具有整體性、系統(tǒng)性、動(dòng)態(tài)性的特點(diǎn)，故，，代謝組學(xué)已廣泛應(yīng)用于疾病、藥物、營養(yǎng)、植物等方面的研究。由于代謝組學(xué)研究的是外源性物質(zhì)對(duì)生物體代謝物組產(chǎn)生的整體效應(yīng)，其思想具有與中醫(yī)藥理論相一致的觀點(diǎn)，利用代謝組學(xué)來研究廣東涼茶這類中草藥作用機(jī)制，將有助于全面了解廣東涼茶的作用機(jī)理。 本研究選擇以大鼠作為研究對(duì)象，探討廣東涼茶在其機(jī)體內(nèi)所產(chǎn)生的代謝組學(xué)規(guī)律。主要研究?jī)?nèi)容如下： 一、基于NMR的代謝組學(xué)研究方法分析不同飲用時(shí)間的大鼠血清樣本，發(fā)現(xiàn)大鼠的代謝組學(xué)響應(yīng)不一樣，表明大鼠對(duì)廣東涼茶的響應(yīng)具有動(dòng)態(tài)性特點(diǎn)。對(duì)于10d飲用廣東涼茶的分析顯現(xiàn)，涼茶組大鼠和對(duì)照組大鼠沒有明顯區(qū)別，這說明大鼠機(jī)體對(duì)廣東涼茶的響應(yīng)不明顯，也可能是大鼠體內(nèi)環(huán)境穩(wěn)定所致。但是，20d的涼茶組大鼠則顯現(xiàn)出與對(duì)照組大鼠明顯的差異，大鼠飲用廣東涼茶后，血糖水平下降，糖異生水平增加，能量代謝由糖代謝途徑轉(zhuǎn)入脂肪代謝。經(jīng)過30d連續(xù)飲用廣東涼茶后，涼茶組大鼠和對(duì)照組大鼠仍然明顯分離，血糖血脂水平降低，但此時(shí)血清中丙酮酸水平下降，表明，廣東涼茶有可能導(dǎo)致其代謝功能低下。 二、利用GC-MS分析技術(shù)和PCA、OPLS-DA等多元數(shù)據(jù)處理方法對(duì)為期38d的涼茶飲用階段里的大鼠尿液代謝物進(jìn)行分析，發(fā)現(xiàn)，反復(fù)飲用廣東涼茶能夠引起體內(nèi)代謝物的積累效應(yīng)和眾多代謝途徑的變化，比如能量代謝、氨基酸代謝等，但是，這些代謝途徑出現(xiàn)的變化趨勢(shì)在整個(gè)響應(yīng)過程中并不一致。因此，對(duì)各個(gè)取樣點(diǎn)的涼茶組大鼠和對(duì)照組大鼠尿液代謝物組進(jìn)行OPLS-DA分析，確定：涼茶組大鼠中前期的主要變化是以抗氧化水平提高、體內(nèi)過氧化水平降低為主要特征；隨著連續(xù)飲用時(shí)間的延長(zhǎng)，這種效應(yīng)慢慢消失，反映了大鼠機(jī)體的自適應(yīng)過程；但進(jìn)一步延長(zhǎng)飲用時(shí)間，大鼠的代謝組響應(yīng)呈現(xiàn)另外一個(gè)特征，即能量流的降低，并出現(xiàn)腸道菌群的紊亂。如果說前期出現(xiàn)的代謝組變化是系統(tǒng)內(nèi)的平衡態(tài)的擾動(dòng)，那末，在后期出現(xiàn)的能量流降低以及腸道微生物菌群的紊亂，可能意味著系統(tǒng)內(nèi)的代謝物組偏離了平衡態(tài)，這種偏離平衡態(tài)的擾動(dòng)表明，長(zhǎng)期連續(xù)飲用廣東涼茶對(duì)大鼠機(jī)體可能導(dǎo)致不利影響。 三、為進(jìn)一步驗(yàn)證廣東涼茶這個(gè)復(fù)雜體系對(duì)大鼠機(jī)體的作用機(jī)制，利用拘束應(yīng)激來模擬“上火”癥狀，采用血清和尿液代謝組學(xué)方法，描述拘束造成的“上火”模型大鼠體內(nèi)的代謝通路變化規(guī)律和廣東涼茶對(duì)拘束應(yīng)激造成大鼠代謝紊亂的恢復(fù)作用，表明，拘束應(yīng)激大鼠給予涼茶后，多數(shù)受到影響的代謝途徑都得到恢復(fù)，其中包括能量代謝、腸道微生物組成的恢復(fù)以及各種涉及機(jī)體免疫力和應(yīng)激能力的氨基酸代謝水平的升高。除此之外，應(yīng)激大鼠在給予涼茶后，機(jī)體抗氧化能力增強(qiáng)，膽固醇水平降低。盡管如此，引起拘束應(yīng)激大鼠中代謝顯著變化的喹啉酸并沒有完全恢復(fù)，表明拘束應(yīng)激造成神經(jīng)系統(tǒng)功能的障礙并未能通過廣東涼茶得到完全改善。
[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.

【學(xué)位授予單位】：華南理工大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2012
【分類號(hào)】：R151

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