Carvalho Rui A, Jarak Ivana
Department of Life Sciences, Faculty of Sciences and Technology, University of Coimbra, Coimbra, Portugal.
Centre for Functional Ecology, University of Coimbra, Coimbra, Portugal.
Methods Mol Biol. 2018;1782:229-247. doi: 10.1007/978-1-4939-7831-1_13.
Metabolic reprogramming has been associated to a plethora of diseases, and there has been increased demand for methodologies able to determine the metabolic alterations that characterize the pathological states and help developing metabolically centered therapies. In this chapter, methodologies for monitoring TCA cycle turnover and its interaction with pyruvate cycling and anaplerotic reactions will be presented. These methodologies are based in the application of stable C isotope "tracers"/substrates and C-NMR isotopomer analysis of metabolic intermediates. These methodologies can be applied at several organizational levels, ranging from isolated organelles and organs to whole organisms/humans. For the sake of simplicity, only very simple and well-defined models will be presented, including isolated heart mitochondria and isolated perfused hearts and livers.
代谢重编程与多种疾病相关，对能够确定表征病理状态的代谢改变并有助于开发以代谢为中心的疗法的方法的需求也在增加。在本章中，将介绍监测三羧酸循环周转及其与丙酮酸循环和回补反应相互作用的方法。这些方法基于稳定碳同位素“示踪剂”/底物的应用以及代谢中间体的碳-核磁共振同位素异构体分析。这些方法可应用于从分离的细胞器和器官到整个生物体/人类的多个组织水平。为简单起见，仅介绍非常简单且定义明确的模型，包括分离的心脏线粒体以及分离的灌注心脏和肝脏。
