Eccentric exercise-induced muscle damage impairs muscle glycogen repletion

Five healthy untrained young male subjects were studied before, immediately after, and 10 days after a 45-min bout of eccentric exercise on a cycle ergometer (201 W). The subjects were sedentary at all other times and consumed a eucaloric meat-free diet. Needle biopsies of the vastus lateralis muscle were examined for intracellular damage and glycogen content. Immediately after exercise, muscle samples showed myofibrillar tearing and edema. At 10 days, there was myofibrillar necrosis, inflammatory cell infiltration, and no evidence of myofibrillar regeneration. Glycogen utilization during the exercise bout was 33 mmol glycosyl units/kg muscle, consistent with the metabolic intensity of 44% of maximal O2 uptake; however, the significant glycogen use by type II fibers contrasted with concentric exercise performed at this intensity. At 10 days after exercise, muscle glycogen was still depleted, in both type I and II fibers. It is possible that the alterations in muscle ultrastructures were related to the lack of repletion of muscle glycogen. Damage produced by eccentric exercise was more persistent than previously reported, indicating that more than 10 days may be necessary for recovery of muscle ultrastructure and carbohydrate reserves.     

Effects of postexercise carbohydrate-protein feedings on muscle glycogen restoration.

The purpose of this investigation was to determine the effects of postexercise eucaloric carbohydrate-protein feedings on muscle glycogen restoration after an exhaustive cycle ergometer exercise bout. Seven male collegiate cyclists [age = 25.6 +/- 1.3 yr, height = 180.9 +/- 3.2 cm, wt = 75.4 +/- 4.0 kg, peak oxygen uptake (VO(2 peak)) = 4.20 +/- 0.2 l/min] performed three trials, each separated by 1 wk: 1) 100% alpha-D-glucose [carbohydrate (CHO)], 2) 70% carbohydrate-20% protein (PRO)-10% fat, and 3) 86% carbohydrate-14% amino acid (AA). All feedings were eucaloric, based on 1.0 g. kg body wt(-1). h(-1) of CHO, and administered every 30 min during a 4-h muscle glycogen restoration period in an 18% wt/vol solution. Muscle biopsies were obtained immediately and 4 h after exercise. Blood samples were drawn immediately after the exercise bout and every 0.5 h for 4 h during the restoration period. Increases in muscle glycogen concentrations for the three feedings (CHO, CHO-PRO, CHO-AA) were 118 mmol/kg dry wt; however, no differences among the feedings were apparent. The serum glucose and insulin responses did not differ throughout the restoration period among the three feedings. These results suggest that muscle glycogen restoration does not appear to be enhanced with the addition of proteins or amino acids to an eucaloric CHO feeding after exhaustive cycle exercise.     

Enhanced Glycogen Storage of a Subcellular Hot Spot in Human Skeletal Muscle during Early Recovery from Eccentric Contractions

Unaccustomed eccentric exercise is accompanied by muscle damage and impaired glucose uptake and glycogen synthesis during subsequent recovery. Recently, it was shown that the role and regulation of glycogen in skeletal muscle are dependent on its subcellular localization, and that glycogen synthesis, as described by the product of glycogen particle size and number, is dependent on the time course of recovery after exercise and carbohydrate availability. In the present study, we investigated the subcellular distribution of glycogen in fibers with high (type I) and low (type II) mitochondrial content during post-exercise recovery from eccentric contractions. Analysis was completed on five male subjects performing an exercise bout consisting of 15 x 10 maximal eccentric contractions. Carbohydrate-rich drinks were subsequently ingested throughout a 48 h recovery period and muscle biopsies for analysis included time points 3, 24 and 48 h post exercise from the exercising leg, whereas biopsies corresponding to prior to and at 48 h after the exercise bout were collected from the non-exercising, control leg. Quantitative imaging by transmission electron microscopy revealed an early (post 3 and 24 h) enhanced storage of intramyofibrillar glycogen (defined as glycogen particles located within the myofibrils) of type I fibers, which was associated with an increase in the number of particles. In contrast, late in recovery (post 48 h), intermyofibrillar, intramyofibrillar and subsarcolemmal glycogen in both type I and II fibers were lower in the exercise leg compared with the control leg, and this was associated with a smaller size of the glycogen particles. We conclude that in the carbohydrate-supplemented state, the effect of eccentric contractions on glycogen metabolism depends on the subcellular localization, muscle fiber’s oxidative capacity, and the time course of recovery. The early enhanced storage of intramyofibrillar glycogen after the eccentric contractions may entail important implications for muscle function and fatigue resistance.     

Role of submaximal exercise in promoting creatine and glycogen accumulation in human skeletal muscle.

We examined the effect of glycogen-depleting exercise on subsequent muscle total creatine (TCr) accumulation and glycogen resynthesis during postexercise periods when the diet was supplemented with carbohydrate (CHO) or creatine (Cr) + CHO. Fourteen subjects performed one-legged cycling exercise to exhaustion. Muscle biopsies were taken from the exhausted (Ex) and nonexhausted (Nex) limbs after exercise and after 6 h and 5 days of recovery, during which CHO (CHO group, n = 7) or Cr + CHO (Cr+CHO group, n = 7) supplements were ingested. Muscle TCr concentration ([TCr]) was unchanged in both groups 6 h after supplementation commenced but had increased in the Ex (P < 0.001) and Nex limbs (P < 0.05) of the Cr+CHO group after 5 days. Greater TCr accumulation was achieved in the Ex limbs (P < 0.01) of this group. Glycogen was increased above nonexercised concentrations in the Ex limbs of both groups after 5 days, with the concentration being greater in the Cr+CHO group (P = 0.06). Thus a single bout of exercise enhanced muscle Cr accumulation, and this effect was restricted to the exercised muscle. However, exercise also diminished CHO-mediated insulin release, which may have attenuated insulin-mediated muscle Cr accumulation. Ingesting Cr with CHO also augmented glycogen supercompensation in the exercised muscle.     

Similar changes of gene expression in human skeletal muscle after resistance exercise and multiple fine needle biopsies

Repeated biopsy sampling from one muscle is necessary to investigate muscular adaptation to different forms of exercise as adaptation is thought to be the result of cumulative effects of transient changes in gene expression in response to single exercise bouts. In a crossover study, we obtained four fine needle biopsies from one vastus lateralis muscle of 11 male subjects (25.9 ± 3.8 yr, 179.2 ± 4.8 cm, 76.5 ± 7.0 kg), taken before (baseline), 1, 4, and 24 h after one bout of squatting exercise performed as conventional squatting or as whole body vibration exercise. To investigate if the repeated biopsy sampling has a confounding effect on the observed changes in gene expression, four fine needle biopsies from one vastus lateralis muscle were also taken from 8 male nonexercising control subjects (24.5 ± 3.7 yr, 180.6 ± 1.2 cm, 81.2 ± 1.6 kg) at the equivalent time points. Using RT-PCR, we observed similar patterns of change in the squatting as well as in the control group for the mRNAs of interleukin 6 (IL-6), IL-6 receptor, insulin-like growth factor 1, p21, phosphofructokinase, and glucose transporter in relation to the baseline biopsy. In conclusion, multiple fine needle biopsies obtained from the same muscle region can per se influence the expression of marker genes induced by an acute bout of resistance exercise.     

Glycogen resynthesis and exercise performance with the addition of fenugreek extract (4-hydroxyisoleucine) to post-exercise carbohydrate feeding

The purpose of this study was to determine the effect of adding fenugreek extract (FG) to post-exercise carbohydrate feeding on glycogen resynthesis and subsequent exercise performance in normoglycemic male endurance athletes. A muscle biopsy sample was obtained from the vastus lateralis from subjects prior to exercise for 5 h at 50% of peak cycling power (52.1 +/- 3.3% of VO(2) peak). A second muscle biopsy sample was obtained immediately after exercise. Immediately after and 2 h after the second biopsy subjects ingested either an oral dose of dextrose (GLU) (1.8 g x kg BW(-1)) or GLU with FG containing 1.99 +/- 0.20 mg x kg(-1) 4-hydroxyisoleucine (GLU + FG) in a randomized, cross-over, double blind design. At 4 h post-exercise a third biopsy was taken and subjects received a standardised meal along with FG or a placebo capsule. At 15 h post-exercise subjects underwent their final muscle biopsy before completing a simulated 40 km cycling time trial. There was no difference in muscle glycogen at any time between GLU and GLU + FG. Additionally, 40 km time trial performance was similar for average power output (221 +/- 28 vs. 213 +/- 16 watts) and for time to completion (69.7 +/- 3.7 vs. 70.5 +/- 2.2 min) for the GLU and GLU + FG, respectively. Despite earlier data to the contrary, the present results do not support an effect of fenugreek supplementation on glycogen resynthesis, even though this may have been the result of differences in experimental protocol.     

Muscle glycogen storage postexercise: effect of mode of carbohydrate administration

The primary purpose of this study was to determine whether gastric emptying limits the rate of muscle glycogen storage during the initial 4 h after exercise when a carbohydrate supplement is provided. A secondary purpose was to determine whether liquid (L) and solid (S) carbohydrate (CHO) feedings result in different rates of muscle glycogen storage after exercise. Eight subjects cycled for 2 h on three separate occasions to deplete their muscle glycogen stores. After each exercise bout they received 3 g CHO/kg body wt in L (50% glucose polymer) or S (rice/banana cake) form or by intravenous infusion (I; 20% sterile glucose). The L and S supplements were divided into two equal doses and administered immediately after and 120 min after exercise, whereas the I supplement was administered continuously during the first 235 min of the 240-min recovery period. Blood samples were drawn from an antecubital vein before exercise, during exercise, and throughout recovery. Muscle biopsies were taken from the vastus lateralis immediately after and 120 and 240 min after exercise. Blood glucose and insulin declined during exercise and increased significantly above preexercise levels during recovery in all treatments. The increase in blood glucose during the I treatment, however, was three times greater than during the L or S treatments. The average insulin response of the L treatment (61.7 +/- 4.9 microU/ml) was significantly greater than that of the S treatment (47.5 +/- 4.2 microU/ml) but not that of the I (55.3 +/- 4.5 microU/ml) treatment.(ABSTRACT TRUNCATED AT 250 WORDS)     

The effects of swimming and running regimens on skeletal muscle glycogen in the rat.

Endurance capacity and the effects of different post-exercise states on skeletal muscle glycogen have been studied in rats trained by swimming or running and in sedentary controls. Regular endurance exercise resulted in increased skeletal muscle glycogen stores. A greater depletion was observed in trained animals than in non-trained animals after a training bout or exhaustive exercise. While muscle glycogen levels did not reflect a differential training stimulus (running vs swimming), swimming as a measure of exhaustive exercise was deemed invalid because of the ability of trained swimmers to avoid stenuous exercise by an alteration of swimming pattern.     

The effect of a carbohydrate and protein supplement on resistance exercise performance, hormonal response, and muscle damage

The purpose of this study was to determine whether resistance exercise performance and postexercise muscle damage were altered when consuming a carbohydrate and protein beverage (CHO-PRO; 6.2% and 1.5% concentrations). Thirty-four male subjects (age: 21.5 +/- 1.7 years; height: 177.3 +/- 1.1 cm; weight: 77.2 +/- 2.2 kg) completed 3 sets of 8 repetitions at their 8 repetition maximum to volitional fatigue. The exercise order consisted of the high pull, leg curl, standing overhead press, leg extension, lat pull-down, leg press, and bench press. In a double-blind, posttest-only control group design, subjects consumed 355 ml of either CHO-PRO or placebo (electrolyte and artificial sweetener beverage) 30 minutes prior to exercise, 177 ml immediately prior to exercise, 177 ml halfway through the exercise bout, and 355 ml immediately following the exercise bout. There were no significant differences between groups relative to exercise performance. Cortisol was significantly elevated in the placebo group compared to the CHO-PRO group at 24 hours postexercise. Insulin was significantly elevated immediately pre-exercise, after the fourth lift, immediately postexercise, 1 hour, and 6 hours postexercise in CHO-PRO compared to the placebo group. Myoglobin levels in the placebo group approached significance halfway through the exercise bout and at 1 hour postexercise (p = 0.06 and 0.07, respectively) and were significantly elevated at 6 hours postexercise compared to the CHO-PRO group. Creatine kinase levels were significantly elevated in the placebo group at 24 hours postexercise compared to the CHO-PRO group. The CHO-PRO supplement did not improve performance during a resistance exercise bout, but appeared to reduce muscle damage, as evidenced by the responses of both myoglobin and creatine kinase. These results suggest the use of a CHO-PRO supplement during resistance training to reduce muscle damage and soreness.     

Regional differences in blood flow and oxygen consumption in resting muscle and their relationship during recovery from exhaustive exercise.

This investigation evaluated regional differences in blood flow and oxygen consumption and their relationship in exercised muscle during recovery from exhaustive exercise. Five healthy men performed exhaustive one-legged cycling exercise. Positron emission tomography was used to measure blood flow, oxygen uptake, and oxygen extraction in the quadriceps femoris muscle before and after exercise. Regions of interest included five areas of the muscle (two proximal, one central, and two distal), which were evenly spaced across the muscle. Before exercise, blood flow and oxygen consumption decreased significantly (P < 0.05) in the direction from the proximal to the distal portions; blood flow declined from 2.0 +/- 0.5 to 1.4 +/- 0.3 ml x 100 g-1 x min-1, and oxygen consumption decreased from 0.21 +/- 0.04 to 0.17 +/- 0.02 ml.100 g-1x min-1. In contrast, these gradients in blood flow and oxygen consumption diminished during recovery after exercise. Consequently, there was a positive relationship between changes in blood flow and oxygen consumption in an exercised muscle during recovery after exercise (r = 0.963, P < 0.01). These changes became larger in the direction from proximal to distal portions: blood flow increased from 2.9 +/- 0.7 to 3.9 +/- 0.8 and oxygen consumption from 1.4 +/- 0.1 to 1.8 +/- 0.4 times resting values. These results suggest that hemodynamic variables are heterogeneous within a muscle both at rest and during recovery from exercise and that there is a systematic difference in these variables in the direction from proximal to distal regions within the quadriceps femoris muscle.     

Creatine supplementation does not affect human skeletal muscle glycogen content in the absence of prior exercise.

Due to the current lack of clarity, we examined whether 5 days of dietary creatine (Cr) supplementation per se can influence the glycogen content of human skeletal muscle. Six healthy male volunteers participated in the study, reporting to the laboratory on four occasions to exercise to the point of volitional exhaustion, each after 3 days of a controlled normal habitual dietary intake. After a familiarization visit, participants cycled to exhaustion in the absence of any supplementation (N), and then 2 wk later again they cycled to exhaustion after 5 days of supplementation with simple sugars (CHO). Finally, after a further 2 wk, they again cycled to exhaustion after 5 days of Cr supplementation. Muscle samples were taken at rest before exercise, at the time point of exhaustion in visit 1, and at subsequent visit time of exhaustion. There was a treatment effect on muscle total Cr content in Cr compared with N and CHO supplementation (P < 0.01). Resting muscle glycogen content was elevated above N following CHO (P < 0.05) but not after Cr. At exhaustion following N, glycogen content was no different from CHO and Cr measured at the same time point during exercise. Cr supplementation under conditions of controlled habitual dietary intake had no effect on muscle glycogen content at rest or after exhaustive exercise. We suggest that any Cr-associated increases in muscle glycogen storage are the result of an interaction between Cr supplementation and other mediators of muscle glycogen storage.     

Impaired muscle glycogen resynthesis after eccentric exercise

Eight men performed 10 sets of 10 eccentric contractions of the knee extensor muscles with one leg [eccentrically exercised leg (EL)]. The weight used for this exercise was 120% of the maximal extension strength. After 30 min of rest the subjects performed two-legged cycling [concentrically exercised leg (CL)] at 74% of maximal O2 uptake for 1 h. In the 3 days after this exercise four subjects consumed diets containing 4.25 g CHO/kg body wt, and the remainder were fed 8.5 g CHO/kg. All subjects experienced severe muscle soreness and edema in the quadriceps muscles of the eccentrically exercised leg. Mean (+/- SE) resting serum creatine kinase increased from a preexercise level of 57 +/- 3 to 6,988 +/- 1,913 U/l on the 3rd day of recovery. The glycogen content (mmol/kg dry wt) in the vastus lateralis of CL muscles averaged 90, 395, and 592 mmol/kg dry wt at 0, 24, and 72 h of recovery. The EL muscle, on the other hand, averaged 168, 329, and 435 mmol/kg dry wt at these same intervals. Subjects receiving 8.5 g CHO/kg stored significantly more glycogen than those who were fed 4.3 g CHO/kg. In both groups, however, significantly less glycogen was stored in the EL than in the CL.     

The effect of pedaling frequency on glycogen depletion rates in type I and type II quadriceps muscle fibers during submaximal cycling exercise

This study was conducted to determine whether the pedaling frequency of cycling at a constant metabolic cost contributes to the pattern of fiber-type glycogen depletion. On 2 separate days, eight men cycled for 30 min at approximately 85% of individual aerobic capacity at pedaling frequencies of either 50 or 100 rev.min-1. Muscle biopsy samples (vastus lateralis) were taken immediately prior to and after exercise. Individual fibers were classified as type I (slow twitch), or type II (fast twitch), using a myosin adenosine triphosphatase stain, and their glycogen content immediately prior to and after exercise quantified via microphotometry of periodic acid-Schiff stain. The 30-min exercise bout resulted in a 46% decrease in the mean optical density (D) of type I fibers during the 50 rev.min-1 condition [0.52 (0.07) to 0.28 (0.04) D units; mean (SEM)] which was not different (P > 0.05) from the 35% decrease during the 100 rev.min-1 condition [0.48 (0.04) to 0.31 (0.05) D units]. In contrast, the mean D in type II fibers decreased 49% during the 50 rev.min-1 condition [0.53 (0.06) to 0.27 (0.04) units]. This decrease was greater (P < 0.05) than the 33% decrease observed in the 100 rev.min-1 condition [0.48 (0.04) to 0.32 (0.06) units). In conclusion, cycling at the same metabolic cost at 50 rather than 100 rev.min-1 results in greater type II fiber glycogen depletion. This is attributed to the increased muscle force required to meet the higher resistance per cycle at the lower pedal frequency.(ABSTRACT TRUNCATED AT 250 WORDS)     

Exercise-induced pyruvate dehydrogenase activation is not affected by 7 days of bed rest

To test the hypothesis that physical inactivity impairs the exercise-induced modulation of pyruvate dehydrogenase (PDH), six healthy normally physically active male subjects completed 7 days of bed rest. Before and immediately after the bed rest, the subjects completed an oral glucose tolerance test (OGTT) and a one-legged knee extensor exercise bout [45 min at 60% maximal load (W(max))] with muscle biopsies obtained from vastus lateralis before, immediately after exercise, and at 3 h of recovery. Blood samples were taken from the femoral vein and artery before and after 40 min of exercise. Glucose intake elicited a larger (P ≤ 0.05) insulin response after bed rest than before, indicating glucose intolerance. There were no differences in lactate release/uptake across the exercising muscle before and after bed rest, but glucose uptake after 40 min of exercise was larger (P ≤ 0.05) before bed rest than after. Muscle glycogen content tended to be higher (0.05< P ≤ 0.10) after bed rest than before, but muscle glycogen breakdown in response to exercise was similar before and after bed rest. PDH-E1α protein content did not change in response to bed rest or in response to the exercise intervention. Exercise increased (P ≤ 0.05) the activity of PDH in the active form (PDHa) and induced (P ≤ 0.05) dephosphorylation of PDH-E1α on Ser2?3, Ser2?? and Ser3??, with no difference before and after bed rest. In conclusion, although 7 days of bed rest induced whole body glucose intolerance, exercise-induced PDH regulation in skeletal muscle was not changed. This suggests that exercise-induced PDH regulation in skeletal muscle is maintained in glucose-intolerant (e.g., insulin resistant) individuals.     

Effect of exercise and training on phospholemman phosphorylation in human skeletal muscle

Phospholemman (PLM, FXYD1) is a partner protein and regulator of the Na(+)-K(+)-ATPase (Na(+)-K(+) pump). We explored the impact of acute and short-term training exercise on PLM physiology in human skeletal muscle. A group of moderately trained males (n = 8) performed a 1-h acute bout of exercise by utilizing a one-legged cycling protocol. Muscle biopsies were taken from vastus lateralis at 0 and 63 min (non-exercised leg) and 30 and 60 min (exercised leg). In a group of sedentary males (n = 9), we determined the effect of a 10-day intense aerobic cycle training on Na(+)-K(+)-ATPase subunit expression, PLM phosphorylation, and total PLM expression as well as PLM phosphorylation in response to acute exercise (1 h at ～72% Vo(2peak)). Biopsies were taken at rest, immediately following, and 3 h after an acute exercise bout before and at the conclusion of the 10-day training study. PLM phosphorylation was increased both at Ser(63) and Ser(68) immediately after acute exercise (75%, P < 0.05, and 30%, P < 0.05, respectively). Short-term training had no adaptive effect on PLM phosphorylation at Ser(63) and Ser(68), nor was the total amount of PLM altered posttraining. The protein expressions of α(1)-, α(2)-,and β(1)-subunits of Na(+)-K(+)-ATPase were increased after training (113%, P < 0.05, 49%, P < 0.05, and 27%, P < 0.05, respectively). Whereas an acute bout of exercise increased the phosphorylation of PKCα/βII on Thr(638/641) pre- and posttraining, phosphorylation of PKCζ/λ on Thr(403/410) was increased in response to acute exercise only after the 10-day training. In conclusion, we show that only acute exercise, and not short-term training, increases phosphorylation of PLM on Ser(63) and Ser(68), and data from one-legged cycling indicate that this effect of exercise on PLM phosphorylation is not due to systemic factors. Our results provide evidence that phosphorylation of PLM may play a role in the acute regulation of the Na(+)-K(+)-ATPase response to exercise.     

Low muscle and liver glycogen contents in dogs treated with thyroid hormones

Muscle biopsies for glycogen determinations were taken from dogs before (controls) and after prolonged treatment with thyroid hormones (T4 or T3). The glycogen content in quadriceps femoris was measured before exercise, immediately after its cessation, and during 24h of post-exercise recovery. The effect of thyroxine treatment on the liver glycogen content both at rest and following physical effort was also studied. A marked decrease in the muscle glycogen content determined at rest was found both in T4 and T3-treated dogs in comparison with controls. Physical exercise diminished the muscle glycogen store to similar values in control and thyroid hormone-treated dogs, but the rate of the muscle glycogen utilization during exercise was lower in the latter. The rate of the post-exercise muscle glycogen synthesis was considerably inhibited in thyroid hormone-treated dogs, but 1 hr glucose infusion, applied immediately after cessation of exercise, accelerated the rate of glycogen re-synthesis, so it was close to that in controls without infusion. Thyroxine treatment also affected the liver glycogen store. Both at rest and after physical exercise significantly lower liver glycogen contents were found in T4-treated dogs than in controls.     

A single bout of exercise activates matrix metalloproteinase in human skeletal muscle.

The aims of this study were 1) to characterize changes in matrix metalloproteinase (MMP), endostatin, and vascular endothelial growth factor (VEGF)-A expression in skeletal muscle in response to a single bout of exercise in humans; and 2) to determine if any exchange of endostatin and VEGF-A between circulation and the exercising leg is associated with a change in the tissue expression or plasma concentration of these factors. Ten healthy males performed 65 min of cycle exercise, and muscle biopsies were obtained from the vastus lateralis muscle at rest and immediately and 120 min after exercise. In the muscle biopsies, measurements of mRNA expression levels of MMP-2, MMP-9, MMP-14, and tissue inhibitor of metalloproteinase; VEGF and endostatin protein levels; and MMP activities were performed. Femoral arterial and venous concentrations of VEGF-A and endostatin were determined before, during, and 120 min after exercise. A single bout of exercise increased MMP-9 mRNA and activated MMP-9 protein in skeletal muscle. No measurable increase of endostatin was observed in the skeletal muscle or in plasma following exercise. A concurrent increase in skeletal muscle VEGF-A mRNA and protein levels was induced by exercise, with no signs of peripheral uptake from the circulation. However, a decrease in plasma VEGF-A concentration occurred following exercise. Thus 1) a single bout of exercise activated the MMP system without any resulting change in tissue endostatin protein levels, and 2) the increased VEGF-A protein levels are due to changes in the skeletal muscle tissue itself. Other mechanisms are responsible for the observed exercise-induced decrease in VEGF-A in plasma.