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Salivary Metabolome and Soccer Match: Challenges for Understanding Exercise induced Changes - PubMed

  • ️Tue Jan 01 2019

Salivary Metabolome and Soccer Match: Challenges for Understanding Exercise induced Changes

Erica Pitti et al. Metabolites. 2019.

Abstract

Saliva samples of seventeen soccer players were analyzed by nuclear magnetic resonance before and after an official match. Two different ways of normalizing data are discussed, using total proteins and total metabolite concentrations. Changes in markers related to energy, hydration status, amino acids and other compounds were found. The limits and advantages of using saliva to define the systemic responses to exercise are examined, both in terms of data normalization and interpretation, and the time that the effect lasts in this biofluid, which is shorter to that commonly observed in blood. The heterogeneous nature and different timing of the exercise developed by players also plays an important role in the metabolic changes that can be measured. Our work focuses mainly on three different aspects: The effect that time sampling has on the observed effect, the type of normalization that is necessary to perform in order to cope with changes in water content, and the metabolic response that can be observed using saliva.

Keywords: NMR; energy metabolism; exercise; hydration status; metabolomics; saliva; sport metabolomics.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1

Nuclear magnetic resonance (NMR) spectrum of a saliva sample with the assignment of major metabolites.

Figure 2
Figure 2

The average values of after/before match ratios referring to (A) total proteins (TPWS) and (B) total observed metabolite concentrations (TOMC).

Figure 3
Figure 3

The ratio between post- and pre-game samples for creatinine, urea nitrogen (SUN), and the ratio SUN/creatinine for the game group and the normally hydrated (NH) group. The last includes both no-game and exit groups. ** p < 0.01. p values were obtained between pre- and post-samples of both game and NH groups. The last comprises both the exit and no-game groups (see text).

Figure 4
Figure 4

Principal component analysis (PCA) analysis of players belonging to the game group. (A) The score plot of pre- (blue) and post- samples (red) for field players. The lines connect the pre- and post-samples for each subject. Goalkeeper samples are depicted in green R2X = 0.895, Q2 = 0.699. (B) and (C) The score and loading plots of the game group excluding the goalkeeper. R2X = 0.907, Q2 = 0.715.

Figure 5
Figure 5

(A) Score plot and (C) loading plot for the PCA analysis of pre (blue) and post-samples (red) of the game group (n = 7) using TPWS normalization. The lines connect samples belonging to the same subject (B) Score plot and (D) loading plot for the PCA analysis of pre (blue) and post-samples (red) of the game group (n = 7) using TOMC-normalized data. Abbreviations: Arg, arginine, Gly, glycine, His, Histidine, Ile, isoleucine, O-PE, o-phosphoethanolamine, Phe, phenylalanine, Thr, threonine, Tyr, tyrosine, Val, valine.

Figure 6
Figure 6

Volcano plots of changes in salivary metabolome for the game group (n = 7) using data normalized using TPWS (A) and TOMC (B). Red lines indicate the two threshold of significance (p = 0.01 and p = 0.05). The fold change, FC, was calculated as log2[post]/[pre].

Figure 7
Figure 7

The concentration changes between pre- (left) and post-samples (right) in energy-related metabolites in saliva using TPWS (A) or TOMC (B) data normalization. In A metabolite concentrations are expressed as µM/mg of total proteins, in B as µM/total metabolic output. The boxes denote interquartile range, lines denotes median, whiskers denotes 5th and 95th percentile. * p < 0.05; ** p < 0.01.

Figure 8
Figure 8

Concentration changes between pre- (left) and post- samples (right) in amino acids and related metabolites in saliva using TPWS (A) or TOMC (B) data normalization. In A metabolite concentrations are expressed as µM/mg of total proteins, in B as µM/total metabolic output. The boxes denote interquartile range, lines denotes median, whiskers denotes 5th and 95th percentile. * p < 0.05; ** p < 0.01.

Figure 8
Figure 8

Concentration changes between pre- (left) and post- samples (right) in amino acids and related metabolites in saliva using TPWS (A) or TOMC (B) data normalization. In A metabolite concentrations are expressed as µM/mg of total proteins, in B as µM/total metabolic output. The boxes denote interquartile range, lines denotes median, whiskers denotes 5th and 95th percentile. * p < 0.05; ** p < 0.01.

Figure 9
Figure 9

Other metabolites showing significant changes between pre- (left) and post-samples (right) when normalizing data by TPWS (A) or TOMC (B). In A metabolite concentrations are expressed as µM/mg of total proteins, in B as µM/total metabolic output. The boxes denote interquartile range, lines denotes median, whiskers denotes 5th and 95th percentile. * p < 0.05; ** p < 0.01.

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