DNA methylation changes during a sprint interval exercise performed under normobaric hypoxia or with blood flow restriction: A pilot study in men.

This crossover study evaluated DNA methylation changes in human salivary samples following single sprint interval training sessions performed in hypoxia, with blood flow restriction (BFR), or with gravity-induced BFR. Global DNA methylation levels were evaluated with an enzyme-linked immunosorbent assay. Methylation-sensitive restriction enzymes were used to determine the percentage methylation in a part of the promoter of the gene-inducible nitric oxide synthase (p-iNOS), as well as an enhancer (e-iNOS). Global methylation increased after exercise (p < 0.001; dz = 0.50). A tendency was observed for exercise × condition interaction (p = 0.070). Post hoc analyses revealed a significant increase in global methylation between pre- (7.2 ± 2.6%) and postexercise (10.7 ± 2.1%) with BFR (p = 0.025; dz = 0.69). Methylation of p-iNOS was unchanged (p > 0.05). Conversely, the methylation of e-iNOS increased from 0.6 ± 0.4% to 0.9 ± 0.8% after exercise (p = 0.025; dz = 0.41), independently of the condition (p > 0.05). Global methylation correlated with muscle oxygenation during exercise (r = 0.37, p = 0.042), while e-iNOS methylation showed an opposite association (r = -0.60, p = 0.025). Furthermore, p-iNOS methylation was linked to heart rate (r = 0.49, p = 0.028). Hence, a single sprint interval training increases global methylation in saliva, and adding BFR tends to increase it further. Lower muscle oxygenation is associated with augmented e-iNOS methylation. Finally, increased cardiovascular strain results in increased p-iNOS methylation.

Impact of systemic hypoxia and blood flow restriction on mechanical, cardiorespiratory, and neuromuscular responses to a multiple-set repeated sprint exercise.

Introduction: Repeated sprint cycling exercises (RSE) performed under systemic normobaric hypoxia (HYP) or with blood flow restriction (BFR) are of growing interest. To the best of our knowledge, there is no stringent consensus on the cardiorespiratory and neuromuscular responses between systemic HYP and BFR during RSE. Thus, this study assessed cardiorespiratory and neuromuscular responses to multiple sets of RSE under HYP or with BFR. Methods: According to a crossover design, fifteen men completed RSE (three sets of five 10-s sprints with 20 s of recovery) in normoxia (NOR), HYP, and with bilaterally-cuffed BFR at 45% of resting arterial occlusive pressure during sets in NOR. Power output, cardiorespiratory and neuromuscular responses were assessed. Results: Average peak and mean powers were lower in BFR (dz = 0.87 and dz = 1.23, respectively) and HYP (dz = 0.65 and dz = 1.21, respectively) compared to NOR (p < 0.001). The percentage decrement of power output was greater in BFR (dz = 0.94) and HYP (dz = 0.64) compared to NOR (p < 0.001), as well as in BFR compared to NOR (p = 0.037, dz = 0.30). The percentage decrease of maximal voluntary contraction of the knee extensors after the session was greater in BFR compared to NOR and HYP (p = 0.011, dz = 0.78 and p = 0.027, dz = 0.75, respectively). Accumulated ventilation during exercise was higher in HYP and lower in BFR (p = 0.002, dz = 0.51, and p < 0.001, dz = 0.71, respectively). Peak oxygen consumption was reduced in HYP (p < 0.001, dz = 1.47). Heart rate was lower in BFR during exercise and recovery (p < 0.001, dz = 0.82 and p = 0.012, dz = 0.43, respectively). Finally, aerobic contribution was reduced in HYP compared to NOR (p = 0.002, dz = 0.46) and BFR (p = 0.005, dz = 0.33). Discussion: Thus, this study indicates that power output during RSE is impaired in HYP and BFR and that BFR amplifies neuromuscular fatigue. In contrast, HYP did not impair neuromuscular function but enhanced the ventilatory response along with reduced oxygen consumption.

Effects of short-term repeated sprint training in hypoxia or with blood flow restriction on response to exercise.

This study compared the effects of a brief repeated sprint training (RST) intervention performed with bilateral blood flow restriction (BFR) conditions in normoxia or conducted at high levels of hypoxia on response to exercise. Thirty-nine endurance-trained athletes completed six repeated sprints cycling sessions spread over 2 weeks consisting of four sets of five sprints (10-s maximal sprints with 20-s active recovery). Athletes were assigned to one of the four groups and subjected to a bilateral partial blood flow restriction (45% of arterial occlusion pressure) of the lower limbs during exercise (BFRG), during the recovery (BFRrG), exercised in a hypoxic room simulating hypoxia at FiO2 ≈ 13% (HG) or were not subjected to additional stress (CG). Peak aerobic power during an incremental test, exercise duration, maximal accumulated oxygen deficit and accumulated oxygen uptake (VO2) during a supramaximal constant-intensity test were improved thanks to RST (p < 0.05). No significant differences were observed between the groups (p > 0.05). No further effect was found on other variables including time-trial performance and parameters of the force-velocity relationship (p > 0.05). Thus, peak aerobic power, exercise duration, maximal accumulated oxygen deficit, and VO2 were improved during a supramaximal constant-intensity exercise after six RST sessions. However, combined hypoxic stress or partial BFR did not further increase peak aerobic power.

Muscle Deoxygenation Rates and Reoxygenation Modeling During a Sprint Interval Training Exercise Performed Under Different Hypoxic Conditions.

This study compared the kinetics of muscle deoxygenation and reoxygenation during a sprint interval protocol performed under four modalities: blood flow restriction at 60% of the resting femoral artery occlusive pressure (BFR), gravity-induced BFR (G-BFR), simulated hypoxia (FiO2≈13%, HYP) and normoxia (NOR). Thirteen healthy men performed each session composed of five all-out 30-s efforts interspaced with 4 min of passive recovery. Total work during the exercises was 17 ± 3.4, 15.8 ± 2.9, 16.7 ± 3.4, and 18.0 ± 3.0 kJ for BFR, G-BFR, HYP and NOR, respectively. Muscle oxygenation was continuously measured with near-infrared spectroscopy. Tissue saturation index (TSI) was modelled with a linear function at the beginning of the sprint and reoxygenation during recovery with an exponential function. Results showed that both models were adjusted to the TSI (R2 = 0.98 and 0.95, respectively). Greater deoxygenation rates were observed in NOR compared to BFR (p = 0.028). No difference was found between the conditions for the deoxygenation rates relative to sprint total work (p > 0.05). Concerning reoxygenation, the amplitude of the exponential was not different among conditions (p > 0.05). The time delay of reoxygenation was longer in BFR compared to the other conditions (p < 0.05). A longer time constant was found for G-BFR compared to the other conditions (p < 0.05), and mean response time was longer for BFR and G-BFR. Finally, sprint performance was correlated with faster reoxygenation. Hence, deoxygenation rates were not different between the conditions when expressed relatively to total sprint work. Furthermore, BFR conditions impair reoxygenation: BFR delays and G-BFR slows down reoxygenation.

Effects of Blood Flow Restriction on O2 Muscle Extraction and O2 Pulmonary Uptake Kinetics During Heavy Exercise.

This study aimed to determine the effects of three levels of blood flow restriction (BFR) on V ˙ O 2 and O 2 extraction kinetics during heavy cycling exercise transitions. Twelve healthy trained males completed two bouts of 10 min heavy intensity exercise without BFR (CON), with 40% or 50% BFR (BFR40 and BFR50, respectively). V ˙ O 2 and tissue saturation index (TSI) were continuously measured and modelled using multiexponential functions. The time constant of the V ˙ O 2 primary phase was significantly slowed in BFR40 (26.4 ± 2.0s; p < 0.001) and BFR50 (27.1 ± 2.1s; p = 0.001) compared to CON (19.0 ± 1.1s). The amplitude of the V ˙ O 2 slow component was significantly increased (p < 0.001) with BFR in a pressure-dependent manner 3.6 ± 0.7, 6.7 ± 0.9 and 9.7 ± 1.0 ml·min-1·kg-1 for CON, BFR40, and BFR50, respectively. While no acceleration of the primary component of the TSI kinetics was observed, there was an increase (p < 0.001) of the phase 3 amplitude with BFR (CON -0.8 ± 0.3% VS BFR40 -2.9 ± 0.9%, CON VS BFR50 -2.8 ± 0.8%). It may be speculated that BFR applied during cycling exercise in the heavy intensity domain shifted the working muscles to an O 2 dependent situation. The acceleration of the extraction kinetics could have reached a plateau, hence not permitting compensation for the slowdown of the blood flow kinetics, and slowing V ˙ O 2 kinetics.

Mechanical, Cardiorespiratory, and Muscular Oxygenation Responses to Sprint Interval Exercises Under Different Hypoxic Conditions in Healthy Moderately Trained Men.

Objective: The aim of this study was to determine the effects of sprint interval exercises (SIT) conducted under different conditions (hypoxia and blood flow restriction [BFR]) on mechanical, cardiorespiratory, and muscular O2 extraction responses. Methods: For this purpose, 13 healthy moderately trained men completed five bouts of 30 s all-out exercises interspaced by 4 min resting periods with lower limb bilateral BFR at 60% of the femoral artery occlusive pressure (BFR60) during the first 2 min of recovery, with gravity-induced BFR (pedaling in supine position; G-BFR), in a hypoxic chamber (FiO2≈13%; HYP) or without additional stress (NOR). Peak and average power, time to achieve peak power, rating of perceived exertion (RPE), and a fatigue index (FI) were analyzed. Gas exchanges and muscular oxygenation were measured by metabolic cart and NIRS, respectively. Heart rate (HR) and peripheral oxygen saturation (SpO2) were continuously recorded. Results: Regarding mechanical responses, peak and average power decreased after each sprint (p < 0.001) excepting between sprints four and five. Time to reach peak power increased between the three first sprints and sprint number five (p < 0.001). RPE increased throughout the exercises (p < 0.001). Of note, peak and average power, time to achieve peak power and RPE were lower in G-BFR (p < 0.001). Results also showed that SpO2 decreased in the last sprints for all the conditions and was lower for HYP (p < 0.001). In addition, Δ[O2Hb] increased in the last two sprints (p < 0.001). Concerning cardiorespiratory parameters, BFR60 application induced a decrease in gas exchange rates, which increased after its release compared to the other conditions (p < 0.001). Moreover, muscle blood concentration was higher for BFR60 (p < 0.001). Importantly, average and peak oxygen consumption and muscular oxyhemoglobin availability during sprints decreased for HYP (p < 0.001). Finally, the tissue saturation index was lower in G-BFR. Conclusions: Thus, SIT associated with G-BFR displayed lower mechanical, cardiorespiratory responses, and skeletal muscle oxygenation than the other conditions. Exercise with BFR60 promotes higher blood accumulation within working muscles, suggesting that BFR60 may additionally affect cellular stress. In addition, HYP and G-BFR induced local hypoxia with higher levels for G-BFR when considering both exercise bouts and recovery periods.

Increase in muscle power is associated with myofibrillar ATPase adaptations during resistance training.

NEW FINDINGS: What is the central question of this study? The aim of this study was to examine the effects of resistance training on gains in the external mechanical power output developed during climbing and myofibrillar ATPase activity in rats. What is the main finding and its importance? Using rapid flow quench experiments, we show that resistance training increases both the power output and the myofibrillar ATPase activity in the flexor digitorum profundus, biceps and deltoid muscles. Data fitting reveals that these functional ameliorations are explained by an increase in the rate constant of liberation of ATP hydrolysis products and contribute to performance gains. ABSTRACT: Skeletal muscle shows a remarkable plasticity that permits functional adaptations in response to different stimulations. To date, modifications of the proportions of myosin heavy chain (MHC) isoforms and increases in fibre size are considered to be the main factors providing sarcomeric plasticity in response to exercise training. In this study, we investigated the effects of a resistance training protocol on the myofibrillar ATPase (m-ATPase) cycle, muscle performance (power output) and MHC gene expression. For this purpose, 8-week-old Wistar Han rats were subjected to 4 weeks of resistance training, with five sessions per week. Muscle samples of flexor digitorum profundus (FDP), biceps and deltoid were collected and subjected to RT-qPCR analyses and assessment of m-ATPase activity with rapid flow quench apparatus. Training led to a significant increase in muscle mass, except for the biceps, and in total mechanical power output (+135.7%, P < 0.001). A shift towards an intermediate fibre type (i.e. MHC2x-to-MHC2a isoform transition) was also observed in biceps and FDP but not in the deltoid muscle. Importantly, rapid flow quench experiments revealed an enhancement of the m-ATPase activity during contraction at maximal velocity (kF ) in the three muscles, with a more marked effect in FDP (+242%, P < 0.001). Data fitting revealed that the rate constant of liberation of ATP hydrolysis products (k3 ) appears to be the main factor influencing the increase in m-ATPase activity. In conclusion, the data showed that, in addition to classically observed changes in MHC isoform content and fibre hypertrophy, m-ATPase activity is enhanced during resistance training and might contribute significantly to performance gains.

AMP-activated protein kinase stabilizes FOXO3 in primary myotubes.

AMP-activated protein kinase (AMPK) is a critical enzyme in conditions of cellular energy deficit such as exercise, hypoxia or nutritional stress. AMPK is well known to regulate protein degradation pathways notably through FOXO-related axis. In this study, we investigated the implication of AMPK activation in FOXO3 expression and stability in skeletal muscle primary myotubes. First, time course and dose response studies revealed optimal AICAR treatment duration and dose in skeletal muscle cells. Then, experiments with cycloheximide treatment of primary myotubes highlighted that AICAR infusion extends FOXO3 protein half-life. Our results also showed that AICAR treatment or nutrient depletion increases FOXO3 expression in primary myotubes and the expression of the mitochondrial E3 ligase Mul1 involved in mitochondrial turnover (mitophagy). In AMPK KO cells, nutrient depletion failed to alter the level of some FOXO3-dependent atrophic genes, including LC3B, BNIP3, and the mitochondrial E3 ligase Mul1, but not the expression of other genes (i.e. FOXO1, Gabarapl1, MAFbx, MuRF1). In summary, our data highlight that AMPK stabilizes FOXO3 and suggest a role in the first initiation step of mitochondrial segregation in muscle cells.

Effects of intermittent hypoxic training performed at high hypoxia level on exercise performance in highly trained runners.

This study exanimated the effects of intermittent hypoxic training (IHT) conducted at a high level of hypoxia with recovery at ambient air on aerobic/anaerobic capacities at sea level and hematological variations. According to a double-blind randomized design, fifteen highly endurance-trained runners completed a 6-weeks regimented training with 3 sessions per week consisting of intermittent runs (6x work-rest ratio of 5':5') on a treadmill at 80-85% of maximal aerobic speed ([Formula: see text]). Nine athletes (hypoxic group, HG) performed the exercise bouts at FI02 = 10.6-11.4% while six athletes (normoxic group, NG) exercised at ambient air. Running time to exhaustion at a velocity corresponding to 95% [Formula: see text] significantly increased for HG while no effect was found for NG. Regarding [Formula: see text], no significant effects were found in either training group. In addition, the decline of jumping performances over a 45s-continuous maximal vertical jump test (i.e. anaerobic capacity index) tended to be lower in HG compared to NG. The levels of the studied hematological variables, including erythropoietin and hematocrit, did not significantly change for either HG or NG. These results highlight that our IHT protocol may induce additional effects on aerobic performance without compromising the anaerobic capacity index in highly-trained athletes.

Regulation of ULK1 Expression and Autophagy by STAT1.

Autophagy involves the lysosomal degradation of cytoplasmic contents for regeneration of anabolic substrates during nutritional or inflammatory stress. Its initiation occurs rapidly after inactivation of the protein kinase mammalian target of rapamycin (mTOR) (or mechanistic target of rapamycin), leading to dephosphorylation of Unc-51-like kinase 1 (ULK1) and autophagosome formation. Recent studies indicate that mTOR can, in parallel, regulate the activity of stress transcription factors, including signal transducer and activator of transcription-1 (STAT1). The current study addresses the role of STAT1 as a transcriptional suppressor of autophagy genes and autophagic activity. We show that STAT1-deficient human fibrosarcoma cells exhibited enhanced autophagic flux as well as its induction by pharmacological inhibition of mTOR. Consistent with enhanced autophagy initiation, ULK1 mRNA and protein levels were increased in STAT1-deficient cells. By chromatin immunoprecipitation, STAT1 bound a putative regulatory sequence in the ULK1 5'-flanking region, the mutation of which increased ULK1 promoter activity, and rendered it unresponsive to mTOR inhibition. Consistent with an anti-apoptotic effect of autophagy, rapamycin-induced apoptosis and cytotoxicity were blocked in STAT1-deficient cells but restored in cells simultaneously exposed to the autophagy inhibitor ammonium chloride. In vivo, skeletal muscle ULK1 mRNA and protein levels as well as autophagic flux were significantly enhanced in STAT1-deficient mice. These results demonstrate a novel mechanism by which STAT1 negatively regulates ULK1 expression and autophagy.

Brain Damage and Motor Cortex Impairment in Chronic Obstructive Pulmonary Disease: Implication of Nonrapid Eye Movement Sleep Desaturation.

STUDY OBJECTIVES: Nonrapid eye movement (NREM) sleep desaturation may cause neuronal damage due to the withdrawal of cerebrovascular reactivity. The current study (1) assessed the prevalence of NREM sleep desaturation in nonhypoxemic patients with chronic obstructive pulmonary disease (COPD) and (2) compared a biological marker of cerebral lesion and neuromuscular function in patients with and without NREM sleep desaturation. METHODS: One hundred fifteen patients with COPD (Global Initiative for Chronic Obstructive Lung Disease [GOLD] grades 2 and 3), resting PaO2 of 60-80 mmHg, aged between 40 and 80 y, and without sleep apnea (apnea-hypopnea index < 15) had polysomnographic sleep recordings. In addition, twenty-nine patients (substudy) were assessed i) for brain impairment by serum S100B (biological marker of cerebral lesion), and ii) for neuromuscular function via motor cortex activation and excitability and maximal voluntary quadriceps strength measurement. RESULTS: A total of 51.3% patients (n = 59) had NREM sleep desaturation (NREMDes). Serum S100B was higher in the NREMDes patients of the substudy (n = 14): 45.1 [Q1: 37.7, Q3: 62.8] versus 32.9 [Q1: 25.7, Q3: 39.5] pg.ml(-1) (P = 0.028). Motor cortex activation and excitability were lower in NREMDes patients (both P = 0.03), but muscle strength was comparable between groups (P = 0.58). CONCLUSIONS: Over half the nonhypoxemic COPD patients exhibited NREM sleep desaturation associated with higher values of the cerebral lesion biomarker and lower neural drive reaching the quadriceps during maximal voluntary contraction. The lack of muscle strength differences between groups suggests a compensatory mechanism(s). Altogether, the results are consistent with an involvement of NREM sleep desaturation in COPD brain impairment. CLINICAL TRIAL REGISTRATION: The study was registered at www.clinicaltrials.gov as NCT01679782.