本文關(guān)鍵詞： 白念珠菌 海藻糖 法尼醇 GC-MS HPLC-MS 代謝 含量測定 基因轉(zhuǎn)錄　出處：《第二軍醫(yī)大學(xué)》2010年碩士論文　論文類型：學(xué)位論文

【摘要】：代謝組學(xué)發(fā)展方興未艾,目前代謝組學(xué)的研究已經(jīng)滲透到微生物學(xué)、植物學(xué)、疾病診斷等各大鄰域。本研究就是借助代謝組學(xué)的技術(shù)手段,以白念珠菌為研究對象,研究其能量代謝物的變化及初步研究其產(chǎn)生變化的原因。 白念珠菌是一種雙相型真菌,在人體中,無癥狀時常表現(xiàn)為酵母細(xì)胞型；侵犯組織和出現(xiàn)癥狀時常表現(xiàn)菌絲型。白念珠菌感染通常是深部感染,致死率比較高。在臨床治療過程中發(fā)現(xiàn)白念珠菌易形成被膜,產(chǎn)生耐藥性,給治療帶來很大阻力。 本論文運(yùn)用代謝指紋分析方法,以GC-MS為分析手段研究白念珠菌的代謝物,區(qū)別浮游型和被膜型白念珠菌,并通過化學(xué)計量學(xué)方法對圖譜進(jìn)行分析,推出兩者可能的差異代謝物。 法尼醇(Farnesol)是抑制白念珠菌由酵母型向菌絲型轉(zhuǎn)變的一種感應(yīng)分子,它由白念珠菌產(chǎn)生并排到細(xì)胞外,研究表明在培養(yǎng)基中外加感應(yīng)分子法尼醇可以抑制白念珠菌生物被膜的形成。 海藻糖(Trehalose)是細(xì)胞中的一種應(yīng)激代謝物,在細(xì)胞處于應(yīng)激時可大量合成,恢復(fù)平常狀態(tài)下又迅速降解。海藻糖含量的變化從某種程度上反映了機(jī)體對外界環(huán)境的響應(yīng)。由于糖的組分復(fù)雜,結(jié)構(gòu)相似,加之糖沒有顯色基團(tuán),難以不經(jīng)衍生就進(jìn)行光譜、色譜分析,因此,糖的分析難度較大。本研究正是建立了一種簡便快速準(zhǔn)確的測定海藻糖的方法,用來測定法尼醇干擾下被膜型白念珠菌中海藻糖的含量變化,結(jié)果發(fā)現(xiàn)法尼醇的干預(yù)確實會引起海藻糖的含量波動。 為了解釋海藻糖含量波動的原因,又從白念珠菌細(xì)胞中海藻糖的代謝循環(huán)著手研究。雖然其代謝循環(huán)過程比較簡單,但卻是由復(fù)雜的調(diào)控機(jī)制所調(diào)控。本文研究調(diào)控代謝過程的酶的基因——hxk、tps1、tps2、ath1、nth、zwf,研究它們在法尼醇作用下基因的轉(zhuǎn)錄水平的變化。研究表明,這六種基因轉(zhuǎn)錄水平差異明顯,部分解釋了海藻糖含量變化的原因。 本課題建立了海藻糖的含量測定方法,研究了白念珠菌被膜形成過程中細(xì)胞內(nèi)法尼醇含量的變化以及外加法尼醇對細(xì)胞內(nèi)海藻糖生成的影響,為白念珠菌耐藥性研究提供了參考。
[Abstract]:The development of metabonomics is in the ascendant. At present, the research of metabonomics has penetrated into the microbiology, botany, disease diagnosis and so on. This study is based on the technical means of metabonomics. The changes of energy metabolites of Candida albicans and their causes were studied. Candida albicans is a biphasic fungus. The infection of Candida albicans is usually deep infection with high mortality. In the course of clinical treatment, Candida albicans are found to be prone to the formation of capsule and drug resistance. Bring a lot of resistance to the treatment. In this paper, the metabolites of Candida albicans were studied by GC-MS, and the phytoplankton and membranous Candida albicans were distinguished, and the chromatogram was analyzed by chemometrics. A possible differential metabolite between the two is suggested. Farnesol is an inductive molecule that inhibits the transformation of Candida albicans from yeast to hyphae. It is produced by Candida albicans and discharged out of cells. The results showed that the addition of faniol could inhibit the formation of biofilm in Candida albicans. Trehalose (Trehalose) is a stress metabolite in cells, which can be synthesized in large quantities when the cells are under stress. The changes of trehalose content reflect the response of the body to the external environment to some extent. The sugar composition is complex, the structure is similar, and the sugar has no chromogenic group. It is difficult to carry out spectrum and chromatographic analysis without derivation, so the analysis of sugar is difficult. This study is to establish a simple, rapid and accurate method for the determination of trehalose. The changes of trehalose content in membranous Candida albicans under the interference of mannitol were determined. The results showed that the intervention of mannitol did cause the fluctuation of trehalose content. In order to explain the fluctuation of trehalose content, the metabolic cycle of trehalose in Candida albicans was studied. However, it is regulated by complex regulatory mechanisms. In this paper, we studied the gene of the enzyme regulating metabolic process, hxktps1, tps2ath1, nththzwf. The results showed that the transcriptional levels of these six genes were significantly different, which partly explained the change of trehalose content. In this paper, a method for the determination of trehalose was established, and the changes of intracellular mannitol content during the formation of Candida albicans membrane and the effect of addition of farnesol on intracellular trehalose production were studied. It provides a reference for the study of resistance of Candida albicans.
【學(xué)位授予單位】：第二軍醫(yī)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2010
【分類號】：R378

【引證文獻(xiàn)】

相關(guān)期刊論文 前1條

1 韋明肯;賴潔玲;詹萍;何盛斌;門杏花;;蝸牛酶高效制備白色念珠菌原生質(zhì)體的研究[J];廣東農(nóng)業(yè)科學(xué);2012年09期

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本文編號：1493678