本文選題：生理狀態(tài)　切入點：溫度　出處：《天津商業(yè)大學》2012年碩士論文　論文類型：學位論文

【摘要】：乳酸鹽并不簡單的只是一種厭氧發(fā)酵產(chǎn)物,或者認為其是造成機體“酸中毒”的罪魁禍首。其實乳酸鹽作為一個調(diào)節(jié)分子,可以整合和調(diào)節(jié)多條代謝途徑。盡管乳酸鹽本身并不具備氧化還原作用,但是它作為一種重要的代謝中間產(chǎn)物參與了糖酵解、生物合成和生物氧化,其在信號調(diào)節(jié)方面的作用體現(xiàn)在調(diào)節(jié)炎癥和抗炎細胞因子,調(diào)節(jié)中心代謝途徑中酶活性等方面。本研究首次對人在基礎(chǔ)狀態(tài)、運動狀態(tài)、學習狀態(tài)、發(fā)燒狀態(tài)下乳酸鹽進行了通量分析和控制分析,主要研究結(jié)果如下:本課題采集四種不同狀態(tài)下志愿者的血液,將分離得到的血清分別進行代謝物變化速率和濃度,以及酶活性的測定。采用熒光-化學發(fā)光檢測儀(Fluoroskan Ascent FL)來進行酶活性測定,測定的原理為酶偶聯(lián)法。運用代謝通量分析和代謝控制分析的方法并結(jié)合主成分分析的方法,評價代謝反應(yīng)步驟對代謝系統(tǒng)的控制程度。研究結(jié)果表明,通過乳酸鹽通量控制分析并結(jié)合主成分分析方法,發(fā)現(xiàn)在最優(yōu)穩(wěn)態(tài)下,丙酮酸脫氫酶系、磷酸果糖激酶、烯醇化酶、丙酮酸激酶和檸檬酸合酶的正調(diào)控作用最大,其中以丙酮酸脫氫酶系CJPDHC=0.246693、烯醇化酶CJENO=0.28504、檸檬酸合酶CJCS=0.164945最為明顯,幾乎占到了通量控制總和的70%。同時,通量控制系數(shù)的分析結(jié)果同樣給出了在通量控制中起負調(diào)控作用的四個酶,分別是CJLDH=-0.22416,CJα-KGDHC=-0.02533,CJG-6-PD=-0.27384,CJMDH=-0.21134。他們分別表示了對乳酸鹽通量起負調(diào)控作用程度的大小。同時,本實驗繪制了不同生理狀態(tài)下人的乳酸鹽通量圖譜,并進行了代謝通量分析。通過對人在不同生理狀態(tài)下乳酸鹽進行通量控制分析,我們確定了對乳酸鹽代謝通量起到關(guān)鍵調(diào)控作用的幾種酶。由于乳酸鹽與癌癥之間有著千絲萬縷的關(guān)系,我們由此可以篩選對乳酸鹽通量起關(guān)鍵調(diào)控作用酶的抑制劑,用以癌癥的治療,或者癌癥手術(shù)愈后保健食品的開發(fā)。
[Abstract]:Lactate is not simply an anaerobic fermentation product, or is thought to be the culprit of the body's "acidosis". In fact, lactate is a regulatory molecule. Although lactate itself does not have redox effect, it is involved in glycolysis, biosynthesis and biological oxidation as an important metabolic intermediate. Its role in signal regulation lies in the regulation of inflammation and anti-inflammatory cytokines, the regulation of enzyme activity in central metabolic pathways, and so on. Flux analysis and control analysis of lactate were carried out under the condition of fever. The main results were as follows: in this study, the blood samples of volunteers in four different states were collected. The enzyme activity was determined by fluorescence chemiluminescence detector Fluoroskan Ascent FLL. The principle of determination was enzyme coupling method. The method of metabolic flux analysis, metabolic control analysis and principal component analysis was used. The results showed that the optimal steady-state stability of pyruvate dehydrogenase system, fructose phosphokinase and enolase were obtained by the analysis of lactate flux control and principal component analysis. The positive regulation of pyruvate kinase and citrate synthase was the greatest, among which pyruvate dehydrogenase was 0.246693, enolase CJENON was 0.28504, citrate synthase CJCS=0.164945 was the most obvious, almost accounted for 70% of the total flux control. The results of flux control coefficient analysis also give four enzymes that play a negative role in flux control, namely, CJLDHU -0.22416 CJ 偽 -KGDHC-0.02533 CJG-6-PDPU -0.27384 CJMDHU -0.21134. They indicate the extent of negative regulation on lactate flux. The flux map of lactate in different physiological states was plotted, and the metabolic flux was analyzed. The flux control of lactate in different physiological state was analyzed. We've identified several enzymes that play a key role in regulating lactate flux. Because of the inextricable relationship between lactate and cancer, we can screen inhibitors that play a key role in lactate flux. For cancer treatment, or for the development of health food after cancer surgery.
【學位授予單位】：天津商業(yè)大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R33

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