本文選題：肝臟 + 氧化應(yīng)激��； 參考：《中國人民解放軍軍事醫(yī)學(xué)科學(xué)院》2006年博士論文

【摘要】：肝臟作為人體的代謝中心，其旺盛的代謝活動必然會伴隨ROS(reactive oxygen species)的大量生成，ROS的大量產(chǎn)生和／或抗氧化防御機制的不足將引起氧化應(yīng)激，另外，酒精、藥物、肥胖引起的過量自由脂肪酸代謝等都極易使肝細(xì)胞產(chǎn)生氧化應(yīng)激，最終引發(fā)肝損傷及肝病。因此，各種氧化應(yīng)激反應(yīng)信號通路及氧化還原調(diào)控蛋白在保護肝臟免受氧化損傷中發(fā)揮重要的作用，但這些蛋白在肝臟中的作用及其作用機制尚無系統(tǒng)性研究。 規(guī)�；鞍踪|(zhì)相互作用研究是揭示蛋白質(zhì)的作用機制，發(fā)現(xiàn)新的蛋白質(zhì)功能、新的調(diào)控途徑以及不同調(diào)控途徑間內(nèi)在聯(lián)系等的重要手段。本文以肝臟氧化應(yīng)激和氧化還原調(diào)控關(guān)鍵蛋白為誘餌，采用高通量酵母雙雜交技術(shù)篩選人肝臟cDNA文庫，從蛋白質(zhì)相互作用角度出發(fā)，對肝臟氧化應(yīng)激反應(yīng)和氧化還原調(diào)控網(wǎng)絡(luò)進行初步研究。 本文成功進行了27個誘餌的文庫篩選，獲得528個候選陽性克隆，除去非編碼序列和不符合讀框的序列，得到223條讀框正確的獵物序列，相對于97對、涉及20個誘餌的相互作用。除11對已知相互作用外，其余為新發(fā)現(xiàn)的相互作用，提示在肝臟氧化應(yīng)激反應(yīng)和氧化還原調(diào)控網(wǎng)絡(luò)中可能還有很多的成員未被發(fā)現(xiàn)。 為了進一步考察得到的相互作用的可靠性，，我們首先對其技術(shù)假陽性和生物學(xué)假陽性進行評價。采用酵母回轉(zhuǎn)驗證實驗對技術(shù)假陽性進行了初步評價，在隨機挑選的61對相互作用中，回轉(zhuǎn)陽性率為52.5％，而已知相互作用的回轉(zhuǎn)陽性率也僅為53.3％。表明我們的數(shù)據(jù)中存在一定的假陽性，同時在回轉(zhuǎn)驗證中存在明顯的假陰性，提示我們不能輕易地去除回轉(zhuǎn)呈陰性的相互作用。對技術(shù)假陽性的有效評價還需要借助其它實驗手段。 在生物學(xué)假陽性評價方面，我們采用多種生物信息分析方法，包括PubMed和HPRD數(shù)據(jù)庫搜索、整合轉(zhuǎn)錄組和基因敲除表型數(shù)據(jù)、GO注釋、相互作用結(jié)構(gòu)域分析、模式生物同源性比較等。發(fā)現(xiàn)我們得到的相互作用中，有22對相互作用(23％)的誘餌和獵物基因名共同出現(xiàn)在相應(yīng)文獻(xiàn)中；26對相互作用(27％)的誘餌和獵物基因表達(dá)相關(guān)；9對相互作用(9％)的誘餌和獵物有相同或相
[Abstract]:As the metabolic center of human body, the vigorous metabolic activity of liver is bound to be accompanied by ROS(reactive oxygen species-based production of Ros and / or deficiency of antioxidant defense mechanism will lead to oxidative stress, in addition, alcohol, drugs, Excessive free fatty acid metabolism caused by obesity can easily lead to oxidative stress in hepatocytes and eventually lead to liver injury and liver disease. Therefore, various oxidative stress signaling pathways and redox regulatory proteins play an important role in protecting the liver from oxidative damage. However, the role of these proteins in the liver and its mechanism have not been systematically studied. The study of large-scale protein interaction is an important means to reveal the mechanism of protein action, to discover new protein functions, new regulatory pathways and internal relations between different regulatory pathways. In this paper, the cDNA library of human liver was screened by high-throughput yeast two-hybrid technique with the key proteins of liver oxidative stress and redox regulation as bait. The oxidative stress response and redox regulatory network of liver were studied. In this paper, 27 decoys were successfully screened, and 528 candidate positive clones were obtained. After removing the non-coding sequence and the unmatched reading frame sequence, the correct prey sequences of 223 reading frames were obtained. Compared with 97 pairs, the interaction of 20 decoys was involved. In addition to 11 pairs of known interactions, the others are newly discovered, suggesting that many members may not be found in the liver oxidative stress response and redox regulatory network. In order to further investigate the reliability of the interaction, we first evaluate the technical false positive and biological false positive. The technical false positive rate was preliminarily evaluated by yeast gyration verification experiment. Among 61 pairs of interactions randomly selected, the positive rate of gyration was 52.5 and the positive rate of known interaction was only 53.3%. It indicates that there are some false positives in our data and obvious false negative in gyration verification, which indicates that we can not easily remove the interaction between gyratory negative and negative. The effective evaluation of false positive technique also needs other experimental means. In the aspect of biological false positive evaluation, we used a variety of biological information analysis methods, including PubMed and HPRD database search, integration of transcription and gene knockout phenotypic data go annotation, interaction domain analysis, model biological homology comparison, and so on. It is found that there are 22 pairs of interaction bait and prey gene name in the corresponding literature.) the bait and prey gene expression are related to the gene expression of prey.
【學(xué)位授予單位】：中國人民解放軍軍事醫(yī)學(xué)科學(xué)院
【學(xué)位級別】：博士
【學(xué)位授予年份】：2006
【分類號】：R341

【引證文獻(xiàn)】

相關(guān)博士學(xué)位論文 前1條

1 周穎;PI3K/Akt通路重要分子的相互作用篩選及初步研究[D];中國人民解放軍軍事醫(yī)學(xué)科學(xué)院;2007年

本文編號：1815445