High Throughput Procedure for Comparative Analysis of In Vivo Cardiac Glucose or Amino Acids Use in Cardiovascular Pathologies and Pharmacological Treatments
Open Access
- 30 July 2021
- journal article
- research article
- Published by MDPI AG in Metabolites
- Vol. 11 (8), 497
- https://doi.org/10.3390/metabo11080497
Abstract
The heart is characterized by the prominent flexibility of its energy metabolism and is able to use diverse carbon substrates, including carbohydrates and amino acids. Cardiac substrate preference could have a major impact on the progress of cardiac pathologies. However, the majority of methods to investigate changes in substrates’ use in cardiac metabolism in vivo are complex and not suitable for high throughput testing necessary to understand and reverse these pathologies. Thus, this study aimed to develop a simple method that would allow for the analysis of cardiac metabolic substrate use. The developed methods involved the subcutaneous injection of stable 13C isotopomers of glucose, valine, or leucine with mass spectrometric analysis for the investigation of its entry into cardiac metabolic pathways that were deducted from 13C alanine and glutamate enrichments in heart extracts. The procedures were validated by confirming the known effects of treatments that modify glucose, free fatty acids, and amino acid metabolism. Furthermore, we studied changes in the energy metabolism of CD73 knock-out mice to demonstrate the potential of our methods in experimental research. The methods created allowed for fast estimation of cardiac glucose and amino acid use in mice and had the potential for high-throughput analysis of changes in pathology and after pharmacological treatments.Funding Information
- National Science Center in Poland (2016/23/B/NZ4/03877 and 2015/17/N/NZ4/028410)
This publication has 51 references indexed in Scilit:
- Targeting fatty acid and carbohydrate oxidation — A novel therapeutic intervention in the ischemic and failing heartBiochimica et Biophysica Acta (BBA) - Molecular Cell Research, 2011
- Incidence rates of heart failure, stroke, and acute myocardial infarction among Type 2 diabetic patients using insulin glargine and other insulinPharmacoepidemiology and Drug Safety, 2009
- Ecto-5′-Nucleotidase Deficiency Exacerbates Pressure-Overload–Induced Left Ventricular Hypertrophy and DysfunctionHypertension, 2008
- The Failing Heart — An Engine Out of FuelThe New England Journal of Medicine, 2007
- Effects of metabolic modulation by trimetazidine on left ventricular function and phosphocreatine/adenosine triphosphate ratio in patients with heart failureEuropean Heart Journal, 2006
- Effects of chronic administration of clenbuterol on function and metabolism of adult rat cardiac muscleAmerican Journal of Physiology-Heart and Circulatory Physiology, 2005
- Electrophysiological Effects of Ranolazine, a Novel Antianginal Agent With Antiarrhythmic PropertiesJournal of the American College of Cardiology, 2004
- Matching coronary blood flow to myocardial oxygen consumptionJournal of Applied Physiology, 2004
- 13C Metabolic Flux AnalysisMetabolic Engineering, 2001
- Metabolism of the heart in health and disease. Part IAmerican Heart Journal, 1968