Six weeks of high-load resistance and low-load blood flow restricted training increase Na/K-ATPase sub-units α2 and ß1 equally, but does not alter ClC-1 abundance in untrained human skeletal muscle.

Contractile function of skeletal muscle relies on the ability of muscle fibers to trigger and propagate action potentials (APs). These electrical signals are created by transmembrane ion transport through ion channels and membrane transporter systems. In this regard, the Cl- ion channel 1 (ClC-1) and the Na+/K--ATPase (NKA) are central for maintaining ion homeostasis across the sarcolemma during intense contractile activity. Therefore, this randomized controlled trial aimed to investigate the changes in ClC-1 and specific NKA subunit isoform expression in response to six weeks (18 training sessions) of high-load resistance exercise (HLRE) and low-load blood flow restricted resistance exercise (BFRRE), respectively. HLRE was conducted as 4 sets of 12 repetitions of knee extensions performed at 70% of 1 repetition maximum (RM), while BFRRE was conducted as 4 sets of knee extensions at 30% of 1RM performed to volitional fatigue. Furthermore, the potential associations between protein expression and contractile performance were investigated. We show that muscle ClC-1 abundance was not affected by either exercise modality, whereas NKA subunit isoforms [Formula: see text]2 and [Formula: see text]1 increased equally by appx. 80-90% with BFRRE (p < 0.05) and 70-80% with HLRE (p < 0.05). No differential impact between exercise modalities was observed. At baseline, ClC-1 protein expression correlated inversely with dynamic knee extensor strength (r=-0.365, p = 0.04), whereas no correlation was observed between NKA subunit content and contractile performance at baseline. However, training-induced changes in NKA [Formula: see text]2 subunit (r = 0.603, p < 0.01) and [Formula: see text]1 subunit (r = 0.453, p < 0.05) correlated with exercise-induced changes in maximal voluntary contraction. These results suggest that the initial adaptation to resistance-based exercise does not involve changes in ClC-1 abundance in untrained skeletal muscle, and that increased content of NKA subunits may facilitate increases in maximal force production.

The Role of Plasma Extracellular Vesicles in Remote Ischemic Conditioning and Exercise-Induced Ischemic Tolerance.

Ischemic conditioning and exercise have been suggested for protecting against brain ischemia-reperfusion injury. However, the endogenous protective mechanisms stimulated by these interventions remain unclear. Here, in a comprehensive translational study, we investigated the protective role of extracellular vesicles (EVs) released after remote ischemic conditioning (RIC), blood flow restricted resistance exercise (BFRRE), or high-load resistance exercise (HLRE). Blood samples were collected from human participants before and at serial time points after intervention. RIC and BFRRE plasma EVs released early after stimulation improved viability of endothelial cells subjected to oxygen-glucose deprivation. Furthermore, post-RIC EVs accumulated in the ischemic area of a stroke mouse model, and a mean decrease in infarct volume was observed for post-RIC EVs, although not reaching statistical significance. Thus, circulating EVs induced by RIC and BFRRE can mediate protection, but the in vivo and translational effects of conditioned EVs require further experimental verification.

Non-failure blood flow restricted exercise induces similar muscle adaptations and less discomfort than failure protocols.

Low-load blood flow restricted resistance exercise (BFRE) performed to volitional failure is suggested to constitute an effective method for producing increases in muscle size and function. However, BFRE to failure may entail high levels of perceived exertion, discomfort, and/or delayed onset of muscle soreness (DOMS). The aim of the study was to compare BFRE performed to volitional failure (F-BFRE) vs non-failure BFRE (NF-BFRE) on changes in muscle size, function and perceptual responses. Fourteen young untrained males had one leg randomized to knee extension F-BFRE while the contralateral leg performed NF-BFRE. The training consisted of 22 exercise bouts over an 8-week period. Whole-muscle cross-sectional area (CSA) of quadriceps components, muscle function, and DOMS were assessed before and after the training period. Perceived exertion and discomfort were registered during each exercise bout. Both F-BFRE and NF-BFRE produced regional increases in muscle CSA in the range of: quadriceps (2.5%-3.8%), vastus lateralis (8.1%-8.5%), and rectus femoris (7.9%-25.0%). All without differences between leg. Muscle strength (6.8%-11.5%) and strength-endurance capacity (13.9%-18.6%) also increased to a similar degree in both legs. Less perceived exertion, discomfort, and DOMS were reported with NF-BFRE compared to F-BFRE. In conclusion, non-failure BFRE enables increases in muscle size and muscle function, while involving reduced perceptions of exertion, discomfort, and DOMS. Non-failure BFRE may be a more feasible approach in clinical settings.

Body position influences arterial occlusion pressure: implications for the standardization of pressure during blood flow restricted exercise.

PURPOSE: Arterial occlusion pressure (AOP) measured in a supine position is often used to set cuff pressures for blood flow restricted exercise (BFRE). However, supine AOP may not reflect seated or standing AOP, thus potentially influencing the degree of occlusion. The primary aim of the study was to investigate the effect of body position on AOP. A secondary aim was to investigate predictors of AOP using wide and narrow cuffs. METHODS: Twenty-four subjects underwent measurements of thigh circumference, skinfold and blood pressure, followed by assessments of thigh AOP in supine and seated positions with a wide and a narrow cuff, respectively, using Doppler ultrasound. RESULTS: In the supine position, AOP was 148 ± 19 and 348 ± 94 mmHg with the wide and narrow cuff, respectively. This increased to 177 ± 20 and 409 ± 101 mmHg in the seated position, with correlations between supine and seated AOP of R 2 = 0.81 and R 2 = 0.50 for the wide and narrow cuff, respectively. For both cuff widths, thigh circumference constituted the strongest predictor of AOP, with diastolic blood pressure explaining additional variance with the wide cuff. The predictive strength of these variables did not differ between body positions. CONCLUSION: Our results indicate that body position strongly influences lower limb AOP, especially with narrow cuffs, yielding very high AOP (≥ 500-600 mmHg) in some subjects. This should be taken into account in the standardization of cuff pressures used during BFRE to better control the physiological effects of BFRE.

Muscle damage and repeated bout effect following blood flow restricted exercise.

PURPOSE: Blood-flow restricted resistance exercise training (BFRE) is suggested to be effective in rehabilitation training, but more knowledge is required about its potential muscle damaging effects. Therefore, we investigated muscle-damaging effects of BFRE performed to failure and possible protective effects of previous bouts of BFRE or maximal eccentric exercise (ECC). METHODS: Seventeen healthy young men were allocated into two groups completing two exercise bouts separated by 14 days. One group performed BFRE in both exercise bouts (BB). The other group performed ECC in the first and BFRE in the second bout. BFRE was performed to failure. Indicators of muscle damage were evaluated before and after exercise. RESULTS: The first bout in the BB group led to decrements in maximum isometric torque, and increases in muscle soreness, muscle water retention, and serum muscle protein concentrations after exercise. These changes were comparable in magnitude and time course to what was observed after first bout ECC. An attenuated response was observed in the repeated exercise bout in both groups. CONCLUSION: We conclude that unaccustomed single-bout BFRE performed to failure induces significant muscle damage. Additionally, both ECC and BFRE can precondition against muscle damage induced by a subsequent bout of BFRE.

Comparative Effects of Aerobic Training and Erythropoietin on Oxygen Uptake in Untrained Humans.

Sieljacks, P, Thams, L, Nellemann, B, Larsen, MS, Vissing, K, and Christensen, B. Comparative effects of aerobic training and erythropoietin on oxygen uptake in untrained humans. J Strength Cond Res 30(8): 2307-2317, 2016-The present study examines responses to 10 weeks of aerobic training and/or erythropoiesis-stimulating agent (ESA) treatment on maximal oxygen uptake (V[Combining Dot Above]O2max). Thirty-six healthy, untrained men were randomly assigned to sedentary-placebo (n = 9), sedentary-ESA (SE) (n = 9), training-placebo (TP) (n = 10), or training-ESA (TE) (n = 8). The participants were treated subcutaneously once weekly with ESA (darbepoietin-α, week 1-3; 40 µg and week 4-10; 20 µg) or a placebo for 10 weeks. The training consisted of supervised cycling 3 times per week for 1 hour at an average of 65% of maximal watt, with a progressive overload during the intervention period. V[Combining Dot Above]O2max, wattmax, and hematological values were measured throughout the study. In addition, the total training workload and estimated energy consumption were recorded after each training session. ESA treatment increased hemoglobin (∼11 and ∼14%, p < 0.001) and hematocrit (∼12 and ∼13%, p < 0.001) in the SE and TE groups, respectively. The relative (but not absolute) increases in V[Combining Dot Above]O2max were more pronounced (p < 0.01) in TE (27 ± 6%), compared with SE (15 ± 4%) but not TP (19 ± 4%), indicating that training is superior to ESA in stimulating V[Combining Dot Above]O2max in untrained men. The increased oxygen uptake in the TE group did not result in higher absolute training workloads than in the TP group. In untrained men, training exhibits a greater stimulus for improvements in V[Combining Dot Above]O2max than ESA treatment, without pronounced additive effects, which is supported by similar average training workloads and energy consumption in TP and TE. Thus, in untrained men, training alone seems sufficient to induce improvement in V[Combining Dot Above]O2max.

Erythropoietin administration alone or in combination with endurance training affects neither skeletal muscle morphology nor angiogenesis in healthy young men.

The aim was to investigate the ability of an erythropoiesis-stimulating agent (ESA), alone or in combination with endurance training, to induce changes in human skeletal muscle fibre and vascular morphology. In a comparative study, 36 healthy untrained men were randomly dispersed into the following four groups: sedentary-placebo (SP, n = 9); sedentary-ESA (SE, n = 9); training-placebo (TP, n = 10); or training-ESA (TE, n = 8). The ESA or placebo was injected once weekly. Training consisted of progressive bicycling three times per week for 10 weeks. Before and after the intervention period, muscle biopsies and magnetic resonance images were collected from the thigh muscles, blood was collected, body composition measured and endurance exercise performance evaluated. The ESA treatment (SE and TE) led to elevated haematocrit, and both ESA treatment and training (SE, TP and TE) increased maximal O2 uptake. With regard to skeletal muscle morphology, TP alone exhibited increases in whole-muscle cross-sectional area and fibre diameter of all fibre types. Also exclusively for TP was an increase in type IIa fibres and a corresponding decrease in type IIx fibres. Furthermore, an overall training effect (TP and TE) was statistically demonstrated in whole-muscle cross-sectional area, muscle fibre diameter and type IIa and type IIx fibre distribution. With regard to muscle vascular morphology, TP and TE both promoted a rise in capillary to muscle fibre ratio, with no differences between the two groups. There were no effects of ESA treatment on any of the muscle morphological parameters. Despite the haematopoietic effects of ESA, we provide novel evidence that endurance training rather than ESA treatment induces adaptational changes in angiogenesis and muscle morphology.

Whole body metabolic effects of prolonged endurance training in combination with erythropoietin treatment in humans: a randomized placebo controlled trial.

UNLABELLED: Erythropoietin (Epo) administration improves aerobic exercise capacity and insulin sensitivity in renal patients and also increases resting energy expenditure (REE). Similar effects are observed in response to endurance training. The aim was to compare the effects of endurance training with erythropoiesis-stimulating agent (ESA) treatment in healthy humans. Thirty-six healthy untrained men were randomized to 10 wk of either: 1) placebo (n = 9), 2) ESA (n = 9), 3) endurance training (n = 10), or 4) ESA and endurance training (n = 8). In a single-blinded design, ESA/placebo was injected one time weekly. Training consisted of biking for 1 h at 65% of wattmax three times per week. Measurements performed before and after the intervention were as follows: body composition, maximal oxygen uptake, insulin sensitivity, REE, and palmitate turnover. Uncoupling protein 2 (UCP2) mRNA levels were assessed in skeletal muscle. Fat mass decreased after training (P = 0.003), whereas ESA induced a small but significant increase in intrahepatic fat (P = 0.025). Serum free fatty acid (FFA) levels and palmitate turnover decreased significantly in response to training, whereas the opposite pattern was found after ESA. REE corrected for lean body mass increased in response to ESA and training, and muscle UCP2 mRNA levels increased after ESA (P = 0.035). Insulin sensitivity increased only after training (P = 0.011). IN CONCLUSION: 1) insulin sensitivity is not improved after ESA treatment despite improved exercise capacity, 2) the calorigenic effects of ESA may be related to increased UCP2 gene expression in skeletal muscle, and 3) training and ESA exert opposite effects on lipolysis under basal conditions, increased FFA levels and liver fat fraction was observed after ESA treatment.

Blood flow-restricted resistance exercise alters the surface profile, miRNA cargo and functional impact of circulating extracellular vesicles.

Ischemic exercise conducted as low-load blood flow restricted resistance exercise (BFRE) can lead to muscle remodelling and promote muscle growth, possibly through activation of muscle precursor cells. Cell activation can be triggered by blood borne extracellular vesicles (EVs) as these nano-sized particles are involved in long distance signalling. In this study, EVs isolated from plasma of healthy human subjects performing a single bout of BFRE were investigated for their change in EV surface profiles and miRNA cargos as well as their impact on skeletal muscle precursor cell proliferation. We found that after BFRE, five EV surface markers and 12 miRNAs were significantly altered. Furthermore, target prediction and functional enrichment analysis of the miRNAs revealed several target genes that are associated to biological pathways involved in skeletal muscle protein turnover. Interestingly, EVs from BFRE plasma increased the proliferation of muscle precursor cells. In addition, alterations in surface markers and miRNAs indicated that the combination of exercise and ischemic conditioning during BFRE can stimulate blood cells to release EVs. These results support that BFRE promotes EV release to engage in muscle remodelling and/or growth processes.

Effect of Blood Flow Restricted Resistance Exercise and Remote Ischemic Conditioning on Functional Capacity and Myocellular Adaptations in Patients With Heart Failure.

BACKGROUND: Patients with congestive heart failure (CHF) have impaired functional capacity and inferior quality of life. The clinical manifestations are associated with structural and functional impairments in skeletal muscle, emphasizing a need for feasible rehabilitation strategies beyond optimal anticongestive medical treatment. We investigated whether low-load blood flow restricted resistance exercise (BFRRE) or remote ischemic conditioning (RIC) could improve functional capacity and quality of life in patients with CHF and stimulate skeletal muscle myofibrillar and mitochondrial adaptations. METHODS: We randomized 36 patients with CHF to BFRRE, RIC, or nontreatment control. BFRRE and RIC were performed 3× per week for 6 weeks. Before and after intervention, muscle biopsies, tests of functional capacity, and quality of life assessments were performed. Deuterium oxide was administered throughout the intervention to measure cumulative RNA and subfraction protein synthesis. Changes in muscle fiber morphology and mitochondrial respiratory function were also assessed. RESULTS: BFRRE improved 6-minute walk test by 39.0 m (CI, 7.0-71.1, P=0.019) compared with control. BFRRE increased maximum isometric strength by 29.7 Nm (CI, 10.8-48.6, P=0.003) compared with control. BFRRE improved quality of life by 5.4 points (CI, -0.04 to 10.9; P=0.052) compared with control. BFRRE increased mitochondrial function by 19.1 pmol/s per milligram (CI, 7.3-30.8; P=0.002) compared with control. RIC did not produce similar changes. CONCLUSIONS: Our results demonstrate that BFRRE, but not RIC, improves functional capacity, quality of life, and muscle mitochondrial function. Our findings have clinical implications for rehabilitation of patients with CHF and provide new insights on the myopathy accompanying CHF. CLINICAL TRIAL REGISTRATION: URL: https://www.clinicaltrials.gov. Unique identifier: NCT03380663.

Six Weeks of Low-Load Blood Flow Restricted and High-Load Resistance Exercise Training Produce Similar Increases in Cumulative Myofibrillar Protein Synthesis and Ribosomal Biogenesis in Healthy Males.

Purpose: High-load resistance exercise contributes to maintenance of muscle mass, muscle protein quality, and contractile function by stimulation of muscle protein synthesis (MPS), hypertrophy, and strength gains. However, high loading may not be feasible in several clinical populations. Low-load blood flow restricted resistance exercise (BFRRE) may provide an alternative approach. However, the long-term protein synthetic response to BFRRE is unknown and the myocellular adaptations to prolonged BFRRE are not well described. Methods: To investigate this, 34 healthy young subjects were randomized to 6 weeks of low-load BFRRE, HLRE, or non-exercise control (CON). Deuterium oxide (D2O) was orally administered throughout the intervention period. Muscle biopsies from m. vastus lateralis were collected before and after the 6-week intervention period to assess long-term myofibrillar MPS and RNA synthesis as well as muscle fiber-type-specific cross-sectional area (CSA), satellite cell content, and myonuclei content. Muscle biopsies were also collected in the immediate hours following single-bout exercise to assess signaling for muscle protein degradation. Isometric and dynamic quadriceps muscle strength was evaluated before and after the intervention. Results: Myofibrillar MPS was higher in BFRRE (1.34%/day, p < 0.01) and HLRE (1.12%/day, p < 0.05) compared to CON (0.96%/day) with no significant differences between exercise groups. Muscle RNA synthesis was higher in BFRRE (0.65%/day, p < 0.001) and HLRE (0.55%/day, p < 0.01) compared to CON (0.38%/day) and both training groups increased RNA content, indicating ribosomal biogenesis in response to exercise. BFRRE and HLRE both activated muscle degradation signaling. Muscle strength increased 6-10% in BFRRE (p < 0.05) and 13-23% in HLRE (p < 0.01). Dynamic muscle strength increased to a greater extent in HLRE (p < 0.05). No changes in type I and type II muscle fiber-type-specific CSA, satellite cell content, or myonuclei content were observed. Conclusions: These results demonstrate that BFRRE increases long-term muscle protein turnover, ribosomal biogenesis, and muscle strength to a similar degree as HLRE. These findings emphasize the potential application of low-load BFRRE to stimulate muscle protein turnover and increase muscle function in clinical populations where high loading is untenable.

Skeletal Muscle Mitochondrial Protein Synthesis and Respiration Increase With Low-Load Blood Flow Restricted as Well as High-Load Resistance Training.

Purpose: It is well established that high-load resistance exercise (HLRE) can stimulate myofibrillar accretion. Additionally, recent studies suggest that HLRE can also stimulate mitochondrial biogenesis and respiratory function. However, in several clinical situations, the use of resistance exercise with high loading may not constitute a viable approach. Low-load blood flow restricted resistance exercise (BFRRE) has emerged as a time-effective low-load alternative to stimulate myofibrillar accretion. It is unknown if BFRRE can also stimulate mitochondrial biogenesis and respiratory function. If so, BFRRE could provide a feasible strategy to stimulate muscle metabolic health. Methods: To study this, 34 healthy previously untrained individuals (24 ± 3 years) participated in BFRRE, HLRE, or non-exercise control intervention (CON) 3 times per week for 6 weeks. Skeletal muscle biopsies were collected; (1) before and after the 6-week intervention period to assess mitochondrial biogenesis and respiratory function and; (2) during recovery from single-bout exercise to assess myocellular signaling events involved in transcriptional regulation of mitochondrial biogenesis. During the 6-week intervention period, deuterium oxide (D2O) was continuously administered to the participants to label newly synthesized skeletal muscle mitochondrial proteins. Mitochondrial respiratory function was assessed in permeabilized muscle fibers with high-resolution respirometry. Mitochondrial content was assessed with a citrate synthase activity assay. Myocellular signaling was assessed with immunoblotting. Results: Mitochondrial protein synthesis rate was higher with BFRRE (1.19%/day) and HLRE (1.15%/day) compared to CON (0.92%/day) (P < 0.05) but similar between exercise groups. Mitochondrial respiratory function increased to similar degree with both exercise regimens and did not change with CON. For instance, coupled respiration supported by convergent electron flow from complex I and II increased 38% with BFRRE and 24% with HLRE (P < 0.01). Training did not alter citrate synthase activity compared to CON. BFRRE and HLRE elicited similar myocellular signaling responses. Conclusion: These results support recent findings that resistance exercise can stimulate mitochondrial biogenesis and respiratory function to support healthy skeletal muscle and whole-body metabolism. Intriquingly, BFRRE produces similar mitochondrial adaptations at a markedly lower load, which entail great clinical perspective for populations in whom exercise with high loading is untenable.

Ten weeks of aerobic training does not result in persistent changes in VLDL triglyceride turnover or oxidation in healthy men.

OBJECTIVE: Very low density lipoprotein triglyceride (VLDL-TG) and free fatty acids (FFA) constitute a substantial proportion of human energy supply both at rest and during exercise. Exercise acutely decreases VLDL-TG concentration, and VLDL-TG clearance is increased after an exercise bout. However, the effects of long-term training are not clear. DESIGN: The aim was to investigate long-term effects of training by direct assessments of VLDL-TG and palmitate kinetics and oxidation in healthy lean men (n=9) at rest, before and after a 10-week training program, compared with a non-training control group (n=9). METHODS: VLDL-TG kinetics were assessed by a primed constant infusion of [1-14C]VLDL-TG, and VLDL-TG oxidation by specific activity (14CO2) in expired air. The metabolic study days were placed 60-72 h after the last exercise bout. RESULTS: Palmitate kinetics and oxidation were assessed by a 2 h constant infusion of [9,10-(3)H]palmitate. In the training group (n=9), maximal oxygen uptake increased significantly by ≈20% (P<0.05), and the insulin sensitivity (assessed by the hyperinsulinemic-euglycemic clamp) improved significantly (P<0.05). Despite these metabolic improvements, no changes were observed in VLDL-TG secretion, clearance, or oxidation or in palmitate kinetics. CONCLUSION: We conclude that 10 weeks of exercise training did not induce changes in VLDL-TG and palmitate kinetics in healthy lean men.