Proceedings of The Physiological Society
University of Oxford (2011) Proc Physiol Soc 23, PC219
Metabolomics by NMR of mice blood and urine after an exhaustive exercise at individual oxidative velocity
L. Le Moyec1, M. Triba2, L. Mille-Hamard1, C. Breuneval1, V. Billat1
1. UBIAE U902 INSERM, University of Evry Val d'essonne, Evry, France. 2. CSPBAT UMR7244, University Paris 13, Bobigny, France.
Metabolomic associates a quantitative analysis method of metabolites contained in biological samples to a multivariate statistical analysis and allows either to establish metabolic profiles as in pathology (obesity, muscular dystrophy, diabetes), or to identify biological markers. The objective of this study is the investigation by 1H NMR spectroscopy of metabolome in mice having performed an exhaustive exercise either at the critical velocity (Vc) which delineate the solicitation of the aerobic metabolism or the peak velocity (Vp). Mice were divided into 3 groups: 10 "control" mice, 7 Vc mice exercising at Vc and 13 Vp mice exercising at Vp, 2h before sacrifice. The total of the exercise was implemented for each mouse. Before sacrifice, blood retrobulbar sampling was performed from anesthetized mice (with an intra-peritoneal injection of pentobarbital (70mg.kg-1) and killed by cervical dislocation). Blood was deproteinized with trichloroacetic acid. The urine was collected, either in the cage before sacrifice, or directly from the bladder after sacrifice. The 1D proton spectra of blood or urine samples were obtained using a Varian spectrometer at 500 MHz. The spectra were divided into 0.001 ppm bins. Their intensities were analyzed by partial least square analysis (PLS) using the Simca-P + software according to their group or to the duration of exercise. Multivariate analysis of blood sample spectra could discriminate the control group from both groups Vc and Vp taken together. The spectra could not be discriminated according to the duration of the exercise. For urine samples, a significant model PLS model could be calculated when comparing the control mice with both groups, Vc and Vpeak. The best model was a PLS model obtained according to the exercise duration with all mice. Taking into account the spectral region having the highest effect on the calculated components of the models, glucose and dimethylamine are characterizing the control mice in blood samples while in urine samples, lactate allantoin, citrate, hippurate and carnitine were the most discriminating metabolites. Metabolic profiles assessed with NMR are highly dependent on the exercise. These results show that urine samples were more informative than blood samples and that the duration of the exercise is a more important parameter to influence the metabolomic status than the velocity of the exercise.
Where applicable, experiments conform with Society ethical requirements