Role of pyruvate dehydrogenase kinase 4 in regulating PDH activation during acute muscle contraction

The oxidation of carbohydrates in mammals is regulated by the pyruvate dehydrogenase (PDH) complex, which is covalently regulated by four PDH kinases (PDK1-4) and two PDH phosphatases (PDP1-2) unique to the PDH complex. To investigate the role that PDK4 plays in regulating PDH activation (PDHa) during muscle contraction, mouse extensor digitorum muscle was removed from wild type (WT) and PDK4-knockout (PDK4-KO) mice after a 24?h fast and stimulated for 3?min either at 10?Hz (low-intensity contraction), 40?Hz (moderate-intensity contraction), or allowed to rest. Force was recorded and muscle PDHa activity and metabolite concentrations were measured. PDHa activity was ～2.5-fold higher at rest in PDK4-KO mice than WT mice (P?=?0.009) and ～2-fold higher in PDK4-KO mice at both 10?Hz (P?相似文献   

Metabolic aspects of low carbohydrate diets and exercise

Following a low carbohydrate diet, there is a shift towards more fat and less carbohydrate oxidation to provide energy to skeletal muscle, both at rest and during exercise. This review summarizes recent work on human skeletal muscle carbohydrate and fat metabolic adaptations to a low carbohydrate diet, focusing mainly on pyruvate dehydrogenase and pyruvate dehydrogenase kinase, and how these changes relate to the capacity for carbohydrate oxidation during exercise.     

Suppression of glycogen consumption during acute exercise by dietary branched-chain amino acids in rats

The effects of a diet supplemented with branched-chain amino acids (BCAA; 4.8% or 6.2%) on BCAA catabolism and glycogen metabolism in rats were examined. Rats were fed a BCAA diet or control diet for 4 wk and part of the rats were subjected to exercise training during the experimental period. Feeding the BCAA diet increased serum BCAA concentrations and activity of the hepatic branched-chain alpha-keto acid dehydrogenase complex, the rate-limiting enzyme in the catabolism of BCAA, suggesting that dietary BCAA promotes BCAA catabolism. Although the serum glucose concentration and glycogen contents in the liver and gastrocnemius muscle of rested rats were not significantly affected by feeding of the BCAA diet, those in rats exhausted by acute exercise were 2-4-fold higher in rats fed the BCAA diet than in rats fed the control diet. The activity of pyruvate dehydrogenase complex in the liver and gastrocnemius muscle after acute exercise showed reverse trends; the complex activities (especially in liver) tended to be less in the BCAA diet group than in the control diet group. These results suggest that dietary BCAA spares glycogen stores in liver and skeletal muscle during exercise and that the decrease in pyruvate dehydrogenase complex activity in these tissues by dietary BCAA is involved in the mechanisms.     

Effects of high-fat diet and physical activity on pyruvate dehydrogenase kinase-4 in mouse skeletal muscle

Background

The expression of PDK4 is elevated by diabetes, fasting and other conditions associated with the switch from the utilization of glucose to fatty acids as an energy source. It is previously shown that peroxisome proliferator-activated receptor ?? coactivator 1?? (PGC-1??), a master regulator of energy metabolism, coactivates in cell lines pyruvate dehydrogenase kinase-4 (PDK4) gene expression via the estrogen-related receptor ?? (ERR??). We investigated the effects of long-term high-fat diet and physical activity on the expression of PDK4, PGC-1?? and ERR?? and the amount and function of mitochondria in skeletal muscle.

Methods

Insulin resistance was induced by a high-fat (HF) diet for 19?weeks in C57BL/6?J mice, which were either sedentary or with access to running wheels. The skeletal muscle expression levels of PDK4, PGC-1?? and ERR?? were measured and the quality and quantity of mitochondrial function was assessed.

Results

The HF mice were more insulin-resistant than the low-fat (LF) -fed mice. Upregulation of PDK4 and ERR?? mRNA and protein levels were seen after the HF diet, and when combined with running even more profound effects on the mRNA expression levels were observed. Chronic HF feeding and voluntary running did not have significant effects on PGC-1?? mRNA or protein levels. No remarkable difference was found in the amount or function of mitochondria.

Conclusions

Our results support the view that insulin resistance is not mediated by the decreased qualitative or quantitative properties of mitochondria. Instead, the role of PDK4 should be contemplated as a possible contributor to high-fat diet-induced insulin resistance.     

Dietary effects on liver and muscle glycogen repletion in exhaustively exercised rats: energy composition and type of complex carbohydrates

Previous reports have indicated that administration of a glucose-citrate (G-C) drink after a bout of exhaustive exercise results in more effective glycogen repletion in liver and skeletal muscle in rats as compared with administration of glucose alone. The present studies report the effects of the energy pattern and the type of carbohydrates, dextrin or starch from rice, in diet given following the G-C drink after exercise, on further glycogen repletion in the tissues of rats. Rats were adapted to meal-feeding 3 times a day and trained with light swimming for 7 to 10 days. On the final day of experiments, rats received the G-C drink after 2 h of exhaustive swimming and were then fed on diets with different energy patterns or carbohydrate types. Results showed that a high-carbohydrate diet is more effective than a high-fat diet for further glycogen repletion in liver and skeletal muscle. In addition, dextrin was revealed to be superior to starch as a carbohydrate source in tissue glycogen repletion. As compared with the high-fat diet, the high-carbohydrate diet, however, resulted in a lower serum free fatty acid concentration 4 h after ingestion of food possibly by decreasing adipose tissue lipolysis.     

What Is the Evidence That Dietary Macronutrient Composition Influences Exercise Performance? A Narrative Review

The introduction of the needle muscle biopsy technique in the 1960s allowed muscle tissue to be sampled from exercising humans for the first time. The finding that muscle glycogen content reached low levels at exhaustion suggested that the metabolic cause of fatigue during prolonged exercise had been discovered. A special pre-exercise diet that maximized pre-exercise muscle glycogen storage also increased time to fatigue during prolonged exercise. The logical conclusion was that the athlete’s pre-exercise muscle glycogen content is the single most important acutely modifiable determinant of endurance capacity. Muscle biochemists proposed that skeletal muscle has an obligatory dependence on high rates of muscle glycogen/carbohydrate oxidation, especially during high intensity or prolonged exercise. Without this obligatory carbohydrate oxidation from muscle glycogen, optimum muscle metabolism cannot be sustained; fatigue develops and exercise performance is impaired. As plausible as this explanation may appear, it has never been proven. Here, I propose an alternate explanation. All the original studies overlooked one crucial finding, specifically that not only were muscle glycogen concentrations low at exhaustion in all trials, but hypoglycemia was also always present. Here, I provide the historical and modern evidence showing that the blood glucose concentration—reflecting the liver glycogen rather than the muscle glycogen content—is the homeostatically-regulated (protected) variable that drives the metabolic response to prolonged exercise. If this is so, nutritional interventions that enhance exercise performance, especially during prolonged exercise, will be those that assist the body in its efforts to maintain the blood glucose concentration within the normal range.     

Human metabolic response to exercise

The energy necessary to support prolonged submaximal exercise is provided by the aerobic metabolism of carbohydrate and fatty acids. Carbohydrate is stored as glycogen, a polymer of glucose, in the liver and in the skeletal muscles, whereas the fatty acids used by working muscles are mainly derived from triglycerides stored in white adipose tissue cells. The relative contributions of carbohydrate and fatty acids to muscle metabolism depend on the relative exercise intensity. The relative exercise intensity is defined as the oxygen cost of the exercise (V0₂) expressed as a percentage of the individual's maximum oxygen uptake (% V0₂ max). At exercise intensities which represent a large % V0₂ max for an individual, muscle glycogen is the main contributor to muscle metabolism. Fatigue is associated with the depletion of the limited intramuscular glycogen stores. When a carbohydrate-rich diet is consumed during recovery after exercise, the muscle glycogen stores are increased above theirpre-exercise concentrations. Thus an exercise and diet regime has been developed to exploit the glycogen supercompensa-tion phenomenon and so increase endurance capacity.     

Energy utilization and repletion during endurance exercise: an historical perspective

Some of the first studies of the effects of diet on exercise performance were reported in the late 1920s. These studies established that eating a high carbohydrate diet before, and ingesting glucose during prolonged exercise delayed the onset of fatigue and enhanced performance, in part by preventing the development of hypoglycaemia. The next significant advance came with the introduction of the muscle biopsy needle which allowed muscle carbohydrate (glycogen) content to be measured for the first time. This technique focused attention on the role of muscle glycogen as possibly the more important factor explaining fatigue during prolonged exercise. Subsequent research inevitably focused on the identification of techniques that would either increase muscle glycogen storage before exercise or decrease the rate of muscle glycogen utilization during subsequent exercise, or both. Coincidentally, studies of fluid balance and of the control of gastric emptying during exercise suggested (a) that dehydration posed a serious health risk especially to marathon runners and (b) that dehydration could only be prevented during prolonged exercise if water alone was ingested. Thus the ‘state of the art’ knowledge in 1970 was that a high carbohydrate diet should be eaten before competition; that muscle glycogen depletion was the main cause of fatigue during prolonged exercise; and that water alone should be ingested in large volumes during exercise. The value of carbohydrate ingestion during exercise was systematically ignored despite the evidence from the classical studies of the 1920s. Fortunately commercial interests came to the rescue of exercise physiology in the mid-1980s. Whereas water is without commercial value, carbohydrate added to the water and ingested by millions of athletes throughout the world has attractive commercial potential. Thus research funded and orchestrated largely by the commercial sector has established that adequate fluid delivery can be provided from drinks containing carbohydrate and that the ingestion of carbohydrate during prolonged exercise can prevent the onset of premature fatigue caused by liver glycogen depletion, hypoglycaemia and a low rate of carbohydrate oxidation. Thus the ‘state of the art’ knowledge in 1990 is that there is no proven value of ingesting water alone during prolonged exercise: that liver glycogen depletion is an important and preventable cause of fatigue during prolonged exercise: and that although muscle glycogen depletion is an important contributor, the real biochemical explanation for fatigue during prolonged exercise is still not clearly established. Nevertheless, there is clear evidence that the provision of exogenous carbohydrate at high rates is essential to delay fatigue and optimize performance during prolonged exercise. It has also become apparent that high rates of both carbohydrate and fluid delivery can be achieved by the adoption of appropriate drinking techniques during exercise.     

Dietary whey protein increases liver and skeletal muscle glycogen levels in exercise-trained rats

We investigated the effect of different types of dietary protein on glycogen content in liver and skeletal muscle of exercise-trained rats. Twenty-four male Sprague-Dawley rats (approximately 100 g; n 6 per group) were divided into sedentary or exercise-trained groups with each group being fed either casein or whey protein as the source of dietary protein. Rats in the exercised groups were trained during 2 weeks using swimming exercise for 120 min/d, 6 d/week. Exercise training resulted in an increase in the skeletal muscle glycogen content. Furthermore, the whey protein group significantly increased the skeletal muscle glycogen content compared with the casein group. The increase in glycogen content in liver was significantly greater in rats fed the whey protein diet compared with those fed the casein diet. We also found that the whey protein diet increased the activity of liver glucokinase, whereas it decreased the activities of 6-phosphofructokinase and pyruvate kinase compared with the casein diet. However, hepatic total glycogen synthase activity and mRNA expression were similar with the two diets. In the skeletal muscle, whey protein decreased only 6-phosphofructokinase activity compared with casein. Total glycogen synthase activity in the skeletal muscle in the whey protein group was significantly higher than that in the casein group. The present study is the first to demonstrate that a diet based on whey protein may increase glycogen content in liver and skeletal muscle of exercise-trained rats. We also observed that whey protein regulated glycogen metabolism in these two tissues by different mechanisms.     

Regulation of glucose metabolism by altered pyruvate dehydrogenase activity. I. Potential site of insulin resistance in sepsis

Regulation of the pyruvate dehydrogenase (PDH) complex has been demonstrated to be a key mechanism in the control of carbohydrate oxidation and conservation of glucose carbon. The effect of sterile inflammation and chronic sepsis (small and large abscess) on the activity of the PDH complex was examined in liver and skeletal muscle. Sepsis altered the proportion of PDH in the active, dephosphorylated form. In hepatic tissue, sterile inflammation leads to a 2.5-fold increase in the proportion of active PDH complex compared to fed control. The same increase in the proportion of active PDH complex was observed in rats with a small septic abscess. However, when the severity of septic episode was increased, the proportion of active PDH complex decreased relative to sterile inflammation or small septic abscess animals. A different pattern in the response to sterile inflammation and sepsis on the proportion of active PDH complex was observed in skeletal muscle compared to liver. In contrast to liver, sterile inflammation did not alter the proportion of active PDH in skeletal muscle. In addition, sepsis (either small or large septic abscess) resulted in a 3-fold decrease in the proportion of active PDH relative to fed control or sterile inflammatory animals. The decrease in the proportion of active PDH complex in sepsis was associated with a corresponding increase in the skeletal muscle acetyl-CoA/CoA ratio. The mechanism responsible for lowered PDH complex activity may have been due to increased PDH kinase activity, secondary to increased skeletal muscle acetyl-CoA/CoA ratios.     

Leucine supplementation enhances skeletal muscle recovery in rats following exercise.

This study was designed to determine the ability of leucine to enhance muscle recovery after exercise. Male rats (200 g) were divided into five groups: sedentary, food-deprived (SF); exercised, food-deprived (EF); exercised, fed a carbohydrate meal (EC); exercised, fed a leucine meal (EL); and exercised, fed a combination of carbohydrate and leucine (ECL). All meals were administered by oral gavage immediately following exercise. EC and ECL meals were isocaloric and provided 15% of daily energy intake. EL and ECL meals each provided 270 mg leucine. Rats ran on a motor-driven treadmill for 2 h at 36 m/min and were killed 1 h postexercise. Plasma glucose and insulin were measured, and the gastrocnemius and plantaris muscles were excised as a unit to determine glycogen levels and the fractional rate of skeletal muscle protein synthesis (Ks). Exercise did not alter plasma glucose or insulin. In contrast, prolonged exercise reduced muscle glycogen (-51%) and Ks (-18%). Refeeding a combination of carbohydrate and leucine increased plasma insulin relative to the EF and SF groups and produced complete recovery of muscle Ks and glycogen to values not different from those in SF rats. Feeding leucine alone restored Ks to that in the SF group without affecting plasma glucose or insulin concentrations. Feeding carbohydrate alone enhanced the rate of glycogen repletion compared to the EF group, concomitant with increases in plasma glucose and insulin. The degree of glycogen recovery correlated with plasma insulin concentrations (r = 0.58, P < 0.05). These data suggest that leucine stimulates muscle protein synthesis following exercise, independent of increased plasma insulin. This is the first demonstration that orally administered leucine stimulates recovery of skeletal muscle protein synthesis after exercise.     

NMR of glycogen in exercise

Natural-abundance 13C NMR spectroscopy is a non-invasive technique that enables in vivo assessments of muscle and/or liver glycogen concentrations. Over the last several years, 13C NMR has been developed and used to obtain information about human glycogen metabolism with diet and exercise. Since NMR is non-invasive, more data points are available over a specified time course, dramatically improving the time resolution. This improved time resolution has enabled the documentation of subtleties of muscle glycogen re-synthesis following severe glycogen depletion that were not previously observed. Muscle and liver glycogen concentrations have been tracked in several different human populations under conditions that include: (1) muscle glycogen recovery from intense localized exercise with normal insulin and with insulin suppressed; (2) muscle glycogen recovery in an insulin-resistant population; (3) muscle glycogen depletion during prolonged low-intensity exercise; (4) effect of a mixed meal on postprandial muscle and liver glycogen synthesis. The present review focuses on basic 13C NMR and gives results from selected studies.     

Effects of Thiamin Restriction on Exercise-Associated Glycogen Metabolism and AMPK Activation Level in Skeletal Muscle

This study aimed to investigate the direct influence of a decrease in the cellular thiamin level, before the onset of anorexia (one of the symptoms of thiamin deficiency) on glycogen metabolism and the AMP-activated protein kinase (AMPK) activation levels in skeletal muscle at rest and in response to exercise. Male Wistar rats were classified as the control diet (CON) group or the thiamin-deficient diet (TD) group and consumed the assigned diets for 1 week. Skeletal muscles were taken from the rats at rest, those that underwent low-intensity swimming (LIS), or high-intensity intermittent swimming (HIS) conducted immediately before dissection. There were no significant differences in food intake, locomotive activity, or body weight between groups, but thiamin pyrophosphate in the skeletal muscles of the TD group was significantly lower than that of the CON group. Muscle glycogen and lactate levels in the blood and muscle were equivalent between groups at rest and in response to exercise. The mitochondrial content was equal between groups, and AMPK in the skeletal muscles of TD rats was normally activated by LIS and HIS. In conclusion, with a lowered cellular thiamin level, the exercise-associated glycogen metabolism and AMPK activation level in skeletal muscle were normally regulated.     

Diet, Exercise, And Endurance

Subjects given a high carbohydrate diet consistently performed better during a prolonged period of heavy exercise than subjects on a "mixed" diet, with the initial level of muscle glycogen determining the quality of performance. It is suggested that high fat     

Dietary whey protein downregulates fatty acid synthesis in the liver, but upregulates it in skeletal muscle of exercise-trained rats

OBJECTIVE: This study compared the effects of casein and whey protein as the source of dietary protein on the activity of lipogenic enzymes and mRNA levels in the liver and skeletal muscle of exercise-trained rats. METHODS: Twenty-eight male Sprague-Dawley rats were randomly assigned to one of four groups (n = 7/group). Rats were assigned to sedentary or exercise-trained groups and were fed the casein or whey protein diet. Rats in the exercise groups were trained for 2 wk using a swimming exercise for 120 min/d and 6 d/wk. RESULTS: A significant decrease in the activity of the hepatic lipogenic enzymes, glucose-6-phosphate dehydrogenase, malic enzyme, adenosine triphosphate citrate lyase, acetyl-coenzyme A carboxylase, and fatty acid synthase (FASN) was observed in rats fed whey protein compared with animals fed casein. Compared with the casein diet, the whey protein diet also lowered mRNA expression of these enzymes, except for FASN. In contrast to the findings in liver, whey protein, as compared with casein, increased skeletal muscle FASN activity and mRNA. Further, exercise training resulted in increased skeletal muscle glucose-6-phosphate dehydrogenase and FASN activity and adenosine triphosphate citrate lyase, acetyl-coenzyme A carboxylase-1, and FASN mRNA expression. CONCLUSIONS: Exercise training or whey protein may play an important role in suppressing hepatic fatty acid synthesis, thereby decreasing accumulation of body fat and stimulating the skeletal muscle to increase energy substrate as fat during prolonged exercise.     

Triacylglycerol infusion improves exercise endurance in patients with mitochondrial myopathy due to complex I deficiency

BACKGROUND: A high-fat diet has been recommended for the treatment of patients with mitochondrial myopathy due to complex I (NADH dehydrogenase) deficiency (CID). OBJECTIVE: This study evaluated the effects of intravenous infusion of isoenergetic amounts of triacylglycerol or glucose on substrate oxidation, glycolytic carbohydrate metabolism, and exercise endurance time and energy state of muscle in CID patients. DESIGN: Four CID patients and 15 control subjects were infused with triacylglycerol (3.7 mg x kg(-1) x min(-1)) or glucose (10 mg x kg(-1) x min(-1)) during low-intensity leg exercise. Respiratory calorimetry was used to evaluate mitochondrial substrate oxidation. The concentration and rate of appearance of plasma lactate (from dilution of [1-(13)C]lactate) were used to evaluate glycolytic carbohydrate metabolism. (31)P magnetic resonance spectroscopy was used to determine ratios of phosphocreatine to inorganic o-phosphate in forearm muscle during exercise. RESULTS: In 3 patients, leg exercise endurance time was better during the triacylglycerol infusion than during the glucose infusion. In all 4 patients, whole-body oxygen consumption rates during exercise were higher during triacylglycerol infusion than during the glucose infusion. In 3 patients, the concentration and rate of appearance of plasma lactate were lower during triacylglycerol infusion than during the glucose infusion. Ratios of phosphocreatine to inorganic o-phosphate during exercise were not significantly different between the 2 infusion studies or between the patients and control subjects. CONCLUSIONS: Triacylglycerol infusion is associated with a greater oxidation of substrates, lower rates of appearance and concentrations of plasma lactate, and greater leg exercise endurance time in myopathic CID patients than is glucose infusion. The energy state of muscle during exercise, however, was not significantly different after infusion of triacylglycerol or glucose.     

Nutrition and exercise determinants of postexercise glycogen synthesis.

During the initial hours of recovery from prolonged exhaustive lower body exercise, muscle glycogen synthesis occurs at rates approximating 1-2 mmol.kg-1 wet wt.hr-1 if no carbohydrate is consumed. When carbohydrate is consumed during the recovery, the maximal rate of glycogen synthesis approximates 7-10 mmol.kg-1 wet wt.hr-1. The rate of post-exercise glycogen synthesis is lower if the magnitude of glycogen degradation is small, if less than 0.7 gm glucose.kg-1 body wt.hr-1 is ingested, when the recovery is active, and when the carbohydrate feeding is delayed. The rate of postexercise glycogen synthesis is not reduced during the initial hours (< 4) after eccentric exercise. For studies evaluating muscle glycogen synthesis in excess of 12 hours of recovery, average rates of glycogen synthesis are below 4 mmol.kg-1 wet wt.hr-1. Glycogen synthesis is known to be impaired for time periods in excess of 24 hours following exercise causing eccentric muscle damage. Following intense exercise resulting in high concentrations of muscle lactate, muscle glycogen synthesis occurs at between 15-25 mmol.kg-1 wet wt.hr-1. These synthesis rates occur without ingested carbohydrate during the recovery period and are maintained when a low intensity active recovery is performed.     

High-fat diet versus habitual diet prior to carbohydrate loading: effects of exercise metabolism and cycling performance

We examined the effects of a high-fat diet (HFD-CHO) versus a habitual diet, prior to carbohydrate (CHO)-loading on fuel metabolism and cycling time-trial (TT) performance. Five endurance-trained cyclists participated in two 14-day randomized cross-over trials during which subjects consumed either a HFD (> 65% MJ from fat) or their habitual diet (CTL) (30 +/- 5% MJ from fat) for 10 day, before ingesting a high-CHO diet (CHO-loading, CHO > 70% MJ) for 3 days. Trials consisted of a 150-min cycle at 70% of peak oxygen uptake (VáO2peak), followed immediately by a 20-km TT. One hour before each trial, cyclists ingested 400 ml of a 3.44% medium-chain triacylglycerol (MCT) solution, and during the trial, ingested 600 ml/hour of a 10% 14C-glucose + 3.44% MCT solution. The dietary treatments did not alter the subjects' weight, body fat, or lipid profile. There were also no changes in circulating glucose, lactate, free fatty acid (FFA), and b-hydroxybutyrate concentrations during exercise. However, mean serum glycerol concentrations were significantly higher (p < .01) in the HFD-CHO trial. The HFD-CHO diet increased total fat oxidation and reduced total CHO oxidation but did not alter plasma glucose oxidation during exercise. By contrast, the estimated rates of muscle glycogen and lactate oxidation were lower after the HFD-CHO diet. The HFD-CHO treatment was also associated with improved TT times (29.5 +/- 2.9 min vs. 30.9 +/- 3.4 min for HFD-CHO and CTL-CHO, p <.05). High-fat feeding for 10 days prior to CHO-loading was associated with an increased reliance on fat, a decreased reliance on muscle glycogen, and improved time trial performance after prolonged exercise.     

Effect of Different Carbohydrate Intakes within 24 Hours after Glycogen Depletion on Muscle Glycogen Recovery in Japanese Endurance Athletes

Daily muscle glycogen recovery after training is important for athletes. Few studies have reported a continuous change in muscle glycogen for 24 h. We aimed to investigate the changes in carbohydrate intake amount on muscle glycogen recovery for 24 h after exercise using ¹³C-magnetic resonance spectroscopy (¹³C-MRS). In this randomized crossover study, eight male participants underwent prolonged high-intensity exercise, and then consumed one of the three carbohydrate meals (5 g/kg body mass (BM)/d, 7 g/kg BM/d, or 10 g/kg BM/d). Glycogen content of thigh muscle was measured using ¹³C-MRS before, immediately after, and 4 h, 12 h and 24 h after exercise. Muscle glycogen concentration decreased to 29.9 ± 15.9% by exercise. Muscle glycogen recovery 4–12 h after exercise for the 5 g/kg group was significantly lower compared to those for 7 g/kg and 10 g/kg groups (p < 0.05). Muscle glycogen concentration after 24 h recovered to the pre-exercise levels for 7 g/kg and 10 g/kg groups; however, there was a significant difference for the 5 g/kg group (p < 0.05). These results suggest that carbohydrate intake of 5 g/kg BM/d is insufficient for Japanese athletes to recover muscle glycogen stores 24 h after completing a long-term high-intensity exercise.