本文關(guān)鍵詞： 束縛應(yīng)激 高同型半胱氨酸血癥 代謝組學(xué) 肝臟 核磁共振 主成分分析法　出處：《華北煤炭醫(yī)學(xué)院》2009年碩士論文　論文類型：學(xué)位論文

【摘要】：目的通過束縛應(yīng)激的建模方法建立大鼠高同型半胱氨酸血癥的模型,探討大鼠高同型半胱氨酸血癥肝臟代謝模式的變化;揭示機(jī)體中同型半胱氨酸含量顯著升高引起肝臟代謝紊亂的相關(guān)作用機(jī)制;尋求代謝標(biāo)志物以及作用靶點(diǎn)。 方法選用健康雄性Wistar大鼠,體重180~200g,隨機(jī)分為對照組和應(yīng)激組。將應(yīng)激組大鼠分為5個應(yīng)激強(qiáng)度不同的時間組,分別束縛1周、2周、4周、6周和8周,建立應(yīng)激動物模型。檢測血漿中的糖皮質(zhì)激素、兒茶酚胺和同型半胱氨酸的含量。 采集模型動物的肝臟組織,采用萃取法分別提取肝臟組織中的水溶性提取物和脂溶性提取物,利用1~H核磁共振技術(shù)對樣品進(jìn)行檢測并通過主成分分析法(PCA)比較正常對照組和應(yīng)激組的大鼠肝臟組織中水溶性提取物和脂溶性提取物的代謝模式,建立穩(wěn)定的肝臟組織代謝組學(xué)的技術(shù)平臺。 通過對模型動物肝臟組織的代謝組學(xué)研究,探討鼠齡和應(yīng)激強(qiáng)度對肝臟代謝狀態(tài)的影響和代謝模式的差異。根據(jù)因子載荷圖(loading plot)和文獻(xiàn)及數(shù)據(jù)庫檢索鑒定出影響代謝模式變化的貢獻(xiàn)量大的代謝小分子。并對這些代謝小分子的代謝途徑進(jìn)行分析,探討代謝小分子在機(jī)體發(fā)生肝臟代謝紊亂時的作用機(jī)制以及與高同型半胱氨酸血癥的相關(guān)性。 結(jié)果通過束縛應(yīng)激的方法建立了高同型半胱氨酸血癥的動物模型。建立了高重復(fù)性和高分辨率的大鼠肝臟組織提取液的代謝組學(xué)的研究方法。通過比較正常和應(yīng)激后肝臟組織1~H-NMR譜圖和PCA分析,發(fā)現(xiàn)不同應(yīng)激強(qiáng)度的大鼠肝臟代謝模式明顯不同并與同鼠齡的對照組大鼠的肝臟代謝模式差異顯著,代謝成分也可明顯區(qū)分開。大鼠肝臟組織水溶性提取物中有24種代謝小分子的含量發(fā)生變化,如乳糖、β-葡萄糖、琥珀酸等;脂溶性提取物中14種代謝小分子的含量發(fā)生變化,如LipidCH3、Lipid (CH2 )n、LipidCH2CH2CO等。這些代謝成分的改變提示束縛應(yīng)激可導(dǎo)致能量代謝中糖酵解、糖異生能力、三羧酸循環(huán)的改變以及肝臟脂代謝的紊亂。其肝臟代謝紊亂可能與與線粒體、內(nèi)質(zhì)網(wǎng)功能降低、氧化磷酸化、脂肪氧化改變以及肝臟抗氧化防御體系受損有關(guān)。 結(jié)論束縛應(yīng)激導(dǎo)致大鼠高同型半胱氨酸血癥的發(fā)生;經(jīng)過1~H-NMR的檢測和PCA分析后發(fā)現(xiàn)束縛應(yīng)激致高同型半胱氨酸血癥大鼠肝臟的代謝狀態(tài)發(fā)生了改變,隨著應(yīng)激強(qiáng)度的增加大鼠肝臟的代謝表型也發(fā)生顯著性變化;經(jīng)鑒定后發(fā)現(xiàn)的對代謝表型改變貢獻(xiàn)量大的代謝小分子(氨基酸、脂溶性化學(xué)結(jié)構(gòu)片段,如谷氨酸、丙氨酸、LipidCH2C=C、LipidCH2CO)可對束縛應(yīng)激導(dǎo)致的高同型半胱氨酸血癥機(jī)體的能量代謝產(chǎn)生巨大影響。
[Abstract]:Objective to establish a rat model of hyperhomocysteinemia by restraint stress and to explore the changes of hepatic metabolic pattern of hyperhomocysteinemia in rats. To reveal the related mechanism of hepatic metabolic disorder caused by the increase of homocysteine content in the body, and to seek for metabolic markers and action targets. Methods healthy male Wistar rats, weighing 180g, were randomly divided into control group and stress group. The rats in the stress group were divided into 5 groups with different stress intensity, one week, two weeks, four weeks, six weeks and eight weeks, respectively. Stress animal model was established. The levels of glucocorticoid, catecholamine and homocysteine in plasma were determined. The liver tissue of the model animal was collected and the water-soluble and fat-soluble extracts from the liver tissue were extracted by extraction method. The metabolic patterns of water-soluble extracts and liposoluble extracts in the liver tissues of rats in normal control group and stress group were compared by using 1H NMR technique and PCAA (principal component analysis). To establish a stable technical platform of liver tissue metabolomics. By studying the metabolomics of liver tissue in model animal, To explore the effect of age and stress intensity on the metabolic state of liver and the difference of metabolic model, small metabolic molecules with large contribution to the change of metabolic pattern were identified according to the factor load map loading plot), literature and database. To analyze the metabolic pathways of these metabolic small molecules, To explore the mechanism of metabolic small molecules in the occurrence of liver metabolic disorder and the correlation with hyperhomocysteinemia. Results the animal model of hyperhomocysteinemia was established by restraint stress. A high reproducibility and high resolution method of metabolomics of rat liver tissue extract was established. 1H-NMR and PCA analysis of liver tissue after stress. It was found that the liver metabolic patterns of rats with different stress intensities were significantly different from those of control rats of the same age. The contents of 24 kinds of metabolic small molecules, such as lactose, 尾 -glucose, succinic acid and so on, were changed in the water-soluble extracts of rat liver, and the contents of 14 kinds of metabolic small molecules in liposoluble extracts were changed. For example, LipidCH3, Lipid CH2, LipidCH2CH2CO and so on. The changes of these metabolic components suggest that restraint stress may lead to glycolysis, glycosylation, changes in tricarboxylic acid cycle and disorder of hepatic lipid metabolism in energy metabolism. Decreased endoplasmic reticulum function, oxidative phosphorylation, lipid oxidation and damage to the liver antioxidant defense system. Conclusion restraint stress leads to the occurrence of hyperhomocysteinemia in rats, and after 1H-NMR detection and PCA analysis, it is found that the metabolic state of liver of rats with hyperhomocysteinemia induced by restraint stress has changed. With the increase of stress intensity, the metabolic phenotype of rat liver also changed significantly, and the small metabolic molecules (amino acids, liposoluble chemical structure fragments, such as glutamic acid, glutamic acid, etc.) that contributed to the change of metabolism phenotype were found after identification. LipidCH2C) has a great effect on the energy metabolism of hyperhomocysteinemia induced by restraint stress.
【學(xué)位授予單位】：華北煤炭醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2009
【分類號】：R363

【參考文獻(xiàn)】

相關(guān)期刊論文 前5條

1 趙劍宇,顏賢忠;基于核磁共振的代謝組學(xué)研究進(jìn)展[J];國外醫(yī)學(xué).藥學(xué)分冊;2004年05期

2 唐惠儒;王玉蘭;;代謝組學(xué):一個迅速發(fā)展的新興學(xué)科(英文)[J];生物化學(xué)與生物物理進(jìn)展;2006年05期

3 楊軍,宋碩林,Jose Castro Perez,Robert S.Plumb,許國旺;代謝組學(xué)及其應(yīng)用[J];生物工程學(xué)報;2005年01期

4 唐惠儒;王玉蘭;;代謝組研究[J];生命科學(xué);2007年03期

5 許彬;王海龍;魏開華;張學(xué)敏;楊松成;;代謝組學(xué)分析技術(shù)及其在幾類重大疾病研究中的應(yīng)用[J];分析測試學(xué)報;2006年05期

，

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