Lifelong Football Training: Effects on Autophagy and Healthy Longevity Promotion

Abstract

Aging is a physiological process characterized by a progressive decline of biological functions and an increase in destructive processes in cells and organs. Physical activity and exercise positively affects the expression of skeletal muscle markers involved in longevity pathways. Recently, a new mechanism, autophagy, was introduced to the adaptations induced by acute and chronic exercise as responsible of positive metabolic modification and health-longevity promotion. However, the molecular mechanisms regulating autophagy in response to physical activity and exercise are sparsely described. We investigated the long-term adaptations resulting from lifelong recreational football training on the expression of skeletal muscle markers involved in autophagy signaling. We demonstrated that lifelong football training increased the expression of messengers: RAD23A, HSPB6, RAB1B, TRAP1, SIRT2, and HSBPB1, involved in the auto-lysosomal and proteasome-mediated protein degradation machinery; of RPL1, RPL4, RPL36, MRLP37, involved in cellular growth and differentiation processes; of the Bcl-2, HSP70, HSP90, PSMD13, and of the ATG5-ATG12 protein complex, involved in proteasome promotion and autophagy processes in muscle samples from lifelong trained subjects compared to age-matched untrained controls. In conclusion, our results indicated that lifelong football training positively influence exercise-induced autophagy processes and protein quality control in skeletal muscle, thus promoting healthy aging.

Keywords: autophagy; cardiovascular capacity; football; lifelong training; longevity.

Figure 1

Clustering output of DEGs in skeletal muscle of Veteran Football Players (VPG) compared to untrained (CG) subjects. In Red are up-regulated and in green down-regulated DEGs. A list of transcript identified, fold-change and gene identification is shows in Supplementary Table S1.

Figure 2

Molecular network generated by the Ingenuity software from the differentially expressed genes in the skeletal muscle from VPG vs CG subjects. The main functional pathways given by Ingenuity for this molecular network are: (A) “Nucleic Acid, amino acid, and protein Metabolism.” Highlights in this network are the RAD23A, HSPB6, HSB1, Rab1B, TRAP1, SIRT2 genes that are all correlated with increase in auto-lysosomal and proteasome-mediated protein degradation machinery and maintenance of protein quality control. (B) “Organ Morphology; Nervous System Development; and Function.” Highlights in this network are ribosomal proteins: RPLP1, RPL4, RPL36, MRLP37 that are involved in cellular growth and proliferation pathways. In red up-regulated and in green down-regulated messenger expression is shown.

Figure 3

Quantitative expression analysis of representative messengers evidenced by IPA analysis in the skeletal muscle from CG and VPG subjects. Quantitative analysis expression (RTqPCR) of RPLP1, RPL4, RPL36, MRPL37, RAB1B, SIRT2, TRAP1, RAD23A, HSPB6, and HSPB1 messenger expression was determined in skeletal muscle biopsies from 15 CG (black bars) to15 VPG subjects (gray bars). An arbitrary value of 1 was assigned to the expression of each messenger in CG. Data represent the means (±SEM) of three different experiments; data were compared using one-way ANOVA and differences were considered significant at ^∗p < 0.05 and ^∗∗p < 0.01 compared to CG.

Figure 4

Effects of lifelong football training on the expression of key markers involved in autophagy. Protein expression levels of HSP70, HSP90, and ATG5-ATG12 complex, involved in autophagy process, of anti-apoptotic Bcl-2, and of PSMD13 subunit of the 19S regulator complex were analyzed by Western Blotting in muscle biopsies from 15 controls (CG, black bars) to15 veterans subjects (VPG, gray bars). GAPDH served as loading control. Representative blots are reported for each protein analyzed. Data were expressed as percentage of CG expression. Comparison between groups was determined by ANOVA and represent the means (±SEM) of three different experiments. Differences were considered significant at ^∗p < 0.05 vs CG.