Milène Catoire

Identification of human exercise-induced myokines using secretome analysis.

Endurance exercise is associated with significant improvements in cardio-metabolic risk parameters. A role for myokines has been hypothesized, yet limited information is available about myokines induced by acute endurance exercise in humans. Therefore, the aim of the study was to identify novel exercise-induced myokines in humans. To this end, we carried out a 1 h one-legged acute endurance exercise intervention in 12 male subjects and a 12 wk exercise training intervention in 18 male subjects. Muscle biopsies were taken before and after acute exercise or exercise training and were subjected to microarray-based analysis of secreted proteins (secretome). For acute exercise, secretome analysis resulted in a list of 86 putative myokines, which was reduced to 29 by applying a fold-change cut-off of 1.5. Based on that shortlist, a selection of putative myokines was measured in the plasma by ELISA or multiplex assay. From that selection, CX3CL1 (fractalkine) and CCL2 (MCP-1) increased at both mRNA and plasma levels. From the known myokines, only IL-6 and FGF 21 changed at the mRNA level, whereas none of the known myokines changed at the plasma level. Secretome analysis of exercise training intervention resulted in a list of 69 putative myokines. Comparing putative myokines altered by acute exercise and exercise training revealed a limited overlap of only 13 genes. In conclusion, this study identified CX3CL1 and CCL2 as myokines that were induced by acute exercise at the gene expression and plasma level and that may be involved in communication between skeletal muscle and other organs.

Pronounced Effects of Acute Endurance Exercise on Gene Expression in Resting and Exercising Human Skeletal Muscle

Regular physical activity positively influences whole body energy metabolism and substrate handling in exercising muscle. While it is recognized that the effects of exercise extend beyond exercising muscle, it is unclear to what extent exercise impacts non-exercising muscles. Here we investigated the effects of an acute endurance exercise bouts on gene expression in exercising and non-exercising human muscle. To that end, 12 male subjects aged 44–56 performed one hour of one-legged cycling at 50% Wmax. Muscle biopsies were taken from the exercising and non-exercising leg before and immediately after exercise and analyzed by microarray. One-legged cycling raised plasma lactate, free fatty acids, cortisol, noradrenalin, and adrenalin levels. Surprisingly, acute endurance exercise not only caused pronounced gene expression changes in exercising muscle but also in non-exercising muscle. In the exercising leg the three most highly induced genes were all part of the NR4A family. Remarkably, many genes induced in non-exercising muscle were PPAR targets or related to PPAR signalling, including PDK4, ANGPTL4 and SLC22A5. Pathway analysis confirmed this finding. In conclusion, our data indicate that acute endurance exercise elicits pronounced changes in gene expression in non-exercising muscle, which are likely mediated by changes in circulating factors such as free fatty acids. The study points to a major influence of exercise beyond the contracting muscle.

Fatty acid-inducible ANGPTL4 governs lipid metabolic response to exercise.

Significance Physical exercise causes profound changes in energy metabolism in humans. In this study we show that resting skeletal muscle has a crucial role in the metabolic response to acute exercise. During endurance exercise, selective induction of the protein angiopoietin-like 4 (ANGPTL4) in nonexercising muscle reduces local fatty acid uptake, presumably to prevent fat overload, while directing fatty acids to the active skeletal muscle as fuel. Our data thus suggest that nonexercising muscle has a key role in governing lipid homeostasis during exercise. Physical activity increases energy metabolism in exercising muscle. Whether acute exercise elicits metabolic changes in nonexercising muscles remains unclear. We show that one of the few genes that is more highly induced in nonexercising muscle than in exercising human muscle during acute exercise encodes angiopoietin-like 4 (ANGPTL4), an inhibitor of lipoprotein lipase-mediated plasma triglyceride clearance. Using a combination of human, animal, and in vitro data, we show that induction of ANGPTL4 in nonexercising muscle is mediated by elevated plasma free fatty acids via peroxisome proliferator-activated receptor-δ, presumably leading to reduced local uptake of plasma triglyceride-derived fatty acids and their sparing for use by exercising muscle. In contrast, the induction of ANGPTL4 in exercising muscle likely is counteracted via AMP-activated protein kinase (AMPK)-mediated down-regulation, promoting the use of plasma triglycerides as fuel for active muscles. Our data suggest that nonexercising muscle and the local regulation of ANGPTL4 via AMPK and free fatty acids have key roles in governing lipid homeostasis during exercise.

The search for exercise factors in humans

Regular exercise reduces the risk for numerous chronic diseases. Exercise not only impacts the contracting skeletal muscle but also elicits systemic changes. The exact mechanisms driving the more systemic changes have yet to be resolved, but exercise factors are thought to be an important missing link. Exercise factors are proteins that are released from skeletal muscle into the circulation during exercise. They represent a subclass of myokines, which are classified as proteins secreted from skeletal muscle serving a signaling role. Here, we provide an overview of the current literature on myokines. Many studies have focused on the identification of new myokines using a variety of approaches. These studies have generated an extensive list of myokines, but so far, the functional relevance of many of these novel myokines remains unclear. Few of these myokines represent putative exercise factors. Currently, IL‐6, secreted protein acidic and rich in cysteine, angiopoietin‐like 4, chemokine (C‐X3‐C motif) ligand 1, and chemokine (C‐C motif) ligand 2 have the highest potential to serve as exercise factors because for all these factors, there is clear evidence that plasma levels increase during exercise. In our view, the future focus should be on characterizing the functional role of myokines in the acute and chronic response to exercise and explore their potential as a target for metabolic diseases.—Catoire, M., Kersten, S. The search for exercise factors in humans. FASEB J. 29, 1615‐1628 (2015). www.fasebj.org

Post-Exercise Rehydration: Effect of Consumption of Beer with Varying Alcohol Content on Fluid Balance after Mild Dehydration.

Purpose The effects of moderate beer consumption after physical activity on rehydration and fluid balance are not completely clear. Therefore, in this study, we investigated the effect of beer consumption, with varying alcohol content, on fluid balance after exercise-induced dehydration. Methods Eleven healthy males were included in this cross over study (age 24.5 ± 4.7 years, body weight 75.4 ± 3.3 kg, VO2max 58.3 ± 6.4 mL kg min−1). Subjects exercised on a cycle ergometer for 45 min at 60% of their maximal power output (Wmax) until mild dehydration (1% body mass loss). Thereafter, in random order, one of five experimental beverages was consumed, in an amount equal to 100% of their sweat loss: non-alcoholic beer (0.0%), low-alcohol beer (2.0%), full-strength beer (5.0%), an isotonic sports drink, and water. Fluid balance was assessed up till 5 h after rehydration. Results After 1 h, urine production was significantly higher for 5% beer compared to the isotonic sports drink (299 ± 143 vs. 105 ± 67 mL; p < 0.01). At the end of the 5-h observation period, net fluid balance (NFB) was negative for all conditions (p = 0.681), with the poorest fluid retention percentage for 5% beer (21% fluid retention) and the best percentage for the isotonic sports drink (42%). Non-alcoholic beer, low-alcoholic beer, and water resulted in fluid retention of 36, 36, and 34%, respectively (p = 0.460). Conclusion There was no difference in NFB between the different beverages. Only a short-lived difference between full-strength beer and the isotonic sports drink in urine output and NFB was observed after mild exercise-induced dehydration. Fluid replacement – either in the form of non-alcoholic beer, low-alcoholic beer, full-strength beer, water, or an isotonic sports drink of 100% of body mass loss was not sufficient to achieve full rehydration. The combination of a moderate amount of beer, with varying alcohol content, enough water or electrolyte- and carbohydrate beverages, and salty foods might improve rehydration, but more research is needed.

PS10 - 1. Fatty acid inducible myokine ANGPTL4 governs the lipid metabolic response to acute exercise

Acute exercise greatly increases cellular demand for energy in active skeletal muscle. During low intensity endurance exercise fatty acids are the major energy source. Fatty acids used during exercise are derived from albumin-bound and triglyceride-derived fatty acid sources.