不同生理状态下人乳酸代谢通量分析和控制分析

发布时间：2018-03-21 08:20

本文选题：生理状态　切入点：温度　出处：《天津商业大学》2012年硕士论文　论文类型：学位论文

【摘要】：乳酸盐并不简单的只是一种厌氧发酵产物,或者认为其是造成机体“酸中毒”的罪魁祸首。其实乳酸盐作为一个调节分子,可以整合和调节多条代谢途径。尽管乳酸盐本身并不具备氧化还原作用,但是它作为一种重要的代谢中间产物参与了糖酵解、生物合成和生物氧化,其在信号调节方面的作用体现在调节炎症和抗炎细胞因子,调节中心代谢途径中酶活性等方面。本研究首次对人在基础状态、运动状态、学习状态、发烧状态下乳酸盐进行了通量分析和控制分析,主要研究结果如下:本课题采集四种不同状态下志愿者的血液,将分离得到的血清分别进行代谢物变化速率和浓度,以及酶活性的测定。采用荧光-化学发光检测仪(Fluoroskan Ascent FL)来进行酶活性测定,测定的原理为酶偶联法。运用代谢通量分析和代谢控制分析的方法并结合主成分分析的方法,评价代谢反应步骤对代谢系统的控制程度。研究结果表明,通过乳酸盐通量控制分析并结合主成分分析方法,发现在最优稳态下,丙酮酸脱氢酶系、磷酸果糖激酶、烯醇化酶、丙酮酸激酶和柠檬酸合酶的正调控作用最大,其中以丙酮酸脱氢酶系CJPDHC=0.246693、烯醇化酶CJENO=0.28504、柠檬酸合酶CJCS=0.164945最为明显,几乎占到了通量控制总和的70%。同时,通量控制系数的分析结果同样给出了在通量控制中起负调控作用的四个酶,分别是CJLDH=-0.22416,CJα-KGDHC=-0.02533,CJG-6-PD=-0.27384,CJMDH=-0.21134。他们分别表示了对乳酸盐通量起负调控作用程度的大小。同时,本实验绘制了不同生理状态下人的乳酸盐通量图谱,并进行了代谢通量分析。通过对人在不同生理状态下乳酸盐进行通量控制分析,我们确定了对乳酸盐代谢通量起到关键调控作用的几种酶。由于乳酸盐与癌症之间有着千丝万缕的关系,我们由此可以筛选对乳酸盐通量起关键调控作用酶的抑制剂,用以癌症的治疗,或者癌症手术愈后保健食品的开发。
[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 伪 -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.
【学位授予单位】：天津商业大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R33

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