Chronic Training Induces Metabolic and Proteomic Response in Male and Female Basketball Players: Salivary Modifications during In-Season Training Programs

Abstract

The aim of this study was to characterize the salivary proteome and metabolome of highly trained female and male young basketball players, highlighting common and different traits. A total of 20 male and female basketball players (10 female and 10 male) and 20 sedentary control subjects (10 female and 10 male) were included in the study. The athletes exercised at least five times per week for 2 h per day. Saliva samples were collected mid-season, between 9:00 and 11:00 a.m. and away from sport competition. The proteome and metabolome were analyzed by using 2DE and GC-MS techniques, respectively. A computerized 2DE gel image analysis revealed 43 spots that varied in intensity among groups. Between these spots, 10 (23.2%) were differentially expressed among male athletes and controls, 22 (51.2%) between female basketball players and controls, 11 spots (25.6%) between male and female athletes, and 13 spots (30.2%) between male and female controls. Among the proteins identified were Immunoglobulin, Alpha-Amylase, and Dermcidin, which are inflammation-related proteins. In addition, several amino acids, such as glutamic acid, lysine, ornithine, glycine, tyrosine, threonine, and valine, were increased in trained athletes. In this study, we highlight that saliva is a useful biofluid to assess athlete performance and confirm that the adaptation of men and women to exercise has some common features, but also some different sex-specific behaviors, including differential amino acid utilization and expression of inflammation-related proteins, which need to be further investigated. Moreover, in the future, it will be interesting to examine the influence of sport-type on these differences.

Keywords: basketball; metabolomics; physical exercise; proteomics; saliva; sex; sport; training.

Figure 1

Proteomic profile of Basketball players and controls. (a) Representative 2DE images of silver-stained gels of saliva proteins run on NL pH 3–10 IP strip and in 9–16% polyacrylamide linear gradient. Circles and numbers indicate statistically differentially abundant proteins between the four groups analyzed, as reported in Table 2. (b) Multivariate analysis of the 2DE gel images results using Principal Components Analysis (PCA) performed by Progenesis SameSpots 4.0 software (Nonlinear Dynamic, UK).

Figure 2

Validation of proteomic results. Histograms and representative immunoblot images of (a) Immunoglobulin A and (b) Alpha-Amylase in BM (basketball male group), CM (control male group), BF (basketball female group), and CF (control female group). Normalization of immunoblot was performed on Coomassie-stained PVDF membrane. The statistical analysis was carried out by two-tailed t-test, using Graphpad Prism 6 (* p < 0.05; ** p < 0.01). (c) Representative image of 2DE silver-stain spots, (d) 2DE Western blot for Dermcidin, and (e) Cystatin A.

Figure 3

Plasma metabolomic profile of female and male basketball players. Histogram representation of saliva metabolites whose relative abundance is statistically different (p < 0.05) between (a) male basketball athletes and controls; (b) female basketball athletes and controls; (c) female and male basketball athletes; and (d) female and male controls (CM, control male group; BF, basketball female group; and CF, control female group). Statistical analysis was performed by two-way ANOVA (Tukey’s multiple comparisons test), using GraphPad Prism 6. Representation of the metabolic pathways involved in (e) male and (f) female in-season chronic training, using the differentially abundant metabolites reported in Table 4. For analysis, we used MetaboAnalyst 4.0, setting a p-value < 0.05 and a false discovery rate (FDR) < 0.1 to select the involved pathways.