Protein metabolism in rat gastrocnemius muscle after stimulated chronic concentric exercise.

Previous results by use of a model of resistance exercise consisting of nonvoluntary electrical contraction of rat skeletal muscle have shown that significant gastrocnemius muscle enlargement was produced after 16 wk of chronic concentric resistance training with progressively increased weights but not after the same training program without weights (J. Appl. Physiol. 65: 950-954, 1988). In the present study we examined whether this differential effect on muscle mass between high- and low-resistance exercise is mediated through differential actions on muscle protein synthesis rates. In addition, we determined whether accumulation of specific mRNA quantities had a primary role in the protein synthesis response to this type of exercise. The data revealed that as little as 8 min of total contractile duration increased gastrocnemius protein synthesis rates by nearly 50%. Contrary to our hypothesis, post-exercise protein synthesis rates do not appear to be differentially regulated by the resistance imposed on the muscle during exercise but rather by the number of repetitions performed during the acute bout. This observation, the failure of high-frequency chronic training to produce gastrocnemius enlargement, and the relatively minor effects on mRNA levels collectively suggest that translational and posttranslational mechanisms, including protein degradation, may be the principal processes by which gastrocnemius protein expression is regulated in this model of stimulated concentric exercise.     

Mitochondrial creatine kinase activity alterations in skeletal muscle during long-distance running

In human gastrocnemius muscle obtained from long-distance runners, mitochondrial creatine kinase (CK) activities were significantly greater than nonrunning control skeletal muscle and significantly increased during training for and after a marathon race. Thus skeletal muscle tended to become similar to heart muscle in its mitochondrial CK composition. Total muscle CK activity was significantly different in males and females, was unaffected by marathon training and racing, and was similar to gastrocnemius muscle obtained from nonrunning controls. There was an inverse correlation between the maximum O2 uptake and the percentage increase in mitochondrial CK activity after training. These studies suggest that mitochondrial CK may play a key role in the intracellular transport of energy from mitochondrial to myofibrils in skeletal muscle during endurance exercise such as long-distance running.     

Effect of endurance training on the phospholipid content of skeletal muscles in the rat

Only few data are available on the effect of training on phospholipid metabolism in skeletal muscles. The aim of the present study was to examine the effect of 6 weeks of endurance training on the content of particular phospholipid fractions and on the incorporation of blood-borne [14C]-palmitic acid into the phospholipids in different skeletal muscles (white and red sections of the gastrocnemius, the soleus and the diaphragm) of the rat. Lipids were extracted from the muscles and separated using thin-layer chromatography into the following fractions: sphingomyelin, phosphatidylcholine, phosphatidylserine, phosphatidylinositol, phosphatidylethanolamine, cardiolipin and neutral lipids (this fraction being composed mostly of triacylglycerols). It was found that training did not affect the content of any phospholipid fraction in soleus muscle. It increased the content of sphingomyelin in white gastrocnemius muscle, cardiolipin and phosphatidylethanolamine in red gastrocnemius muscle and phosphatidylinositol in white gastrocnemius muscle and diaphragm. The total phospholipid content in red gastrocnemius muscle of the trained group was higher than in the control group. Training reduced the specific activity of sphingomyelin and cardiolipin in all muscles, phosphatidylcholine in soleus, red, and white gastrocnemius muscles, phosphatidylserine in all muscles, phosphatidylinositol in all except the soleus muscle, and phosphatidylethanolamine in hindleg muscles, but not in the diaphragm compared to the corresponding values in the sedentary group. It was concluded that endurance training affects skeletal muscle phospholipid content and the rate of incorporation of the blood-borne [14C]palmitic acid into the phospholipid moieties.     

Combined isometric, concentric, and eccentric resistance exercise prevents unloading-induced muscle atrophy in rats.

Previously, we reported that an isometric resistance training program that was effective in stimulating muscle hypertrophy in ambulatory rats could not completely prevent muscle atrophy during unloading (Haddad F, Adams GR, Bodell PW, Baldwin KM. J Appl Physiol 100: 433-441, 2006). These results indicated that preventing muscle atrophy does not appear to be simply a function of providing an anabolic stimulus. The present study was undertaken to determine if resistance training, with increased volume (3-s contractions) and incorporating both static and dynamic components, would be effective in preventing unloading-induced muscle atrophy. Rats were exposed to 5 days of muscle unloading via tail suspension. During that time one leg received electrically stimulated resistance exercise (RE) that included an isometric, concentric, and eccentric phase. The results of this study indicate that this combined-mode RE provided an anabolic stimulus sufficient to maintain the mass and myofibril content of the trained but not the contralateral medial gastrocnemius (MG) muscle. Relative to the contralateral MG, the RE stimulus increased the amount of total RNA (indicative of translational capacity) as well as the mRNA for several anabolic/myogenic markers such as insulin-like growth factor-I, myogenin, myoferlin, and procollagen III-alpha-1 and decreased that of myostatin, a negative regulator of muscle size. The combined-mode RE protocol also increased the activity of anabolic signaling intermediates such as p70S6 kinase. These results indicate that a combination of static- and dynamic-mode RE of sufficient volume provides an effective stimulus to stimulate anabolic/myogenic mechanisms to counter the initial stages of unloading-induced muscle atrophy.     

Effect of resistance exercise training on expression of Hsp70 and inflammatory cytokines in skeletal muscle and adipose tissue of STZ-induced diabetic rats

Impairment of adipose tissue and skeletal muscles accrued following type 1 diabetes is associated with protein misfolding and loss of adipose mass and skeletal muscle atrophy. Resistance training can maintain muscle mass by changing both inflammatory cytokines and stress factors in adipose tissue and skeletal muscle. The purpose of this study was to determine the effects of a 5-week ladder climbing resistance training program on the expression of Hsp70 and inflammatory cytokines in adipose tissue and fast-twitch flexor hallucis longus (FHL) and slow-twitch soleus muscles in healthy and streptozotocin-induced diabetic rats. Induction of diabetes reduced body mass, while resistance training preserved FHL muscle weight in diabetic rats without any changes in body mass. Diabetes increased Hsp70 protein content in skeletal muscles, adipose tissue, and serum. Hsp70 protein levels were decreased in normal and diabetic rats by resistance training in the FHL, but not soleus muscle. Furthermore, resistance training decreased inflammatory cytokines in FHL skeletal muscle. On the other hand, Hsp70 and inflammatory cytokine protein levels were increased by training in adipose tissue. Also, significant positive correlations between inflammatory cytokines in adipose tissue and skeletal muscles with Hsp70 protein levels were observed. In conclusion, we found that in diabetic rats, resistance training decreased inflammatory cytokines and Hsp70 protein levels in fast skeletal muscle, increased adipose tissue inflammatory cytokines and Hsp70, and preserved FHL muscle mass. These results suggest that resistance training can maintain skeletal muscle mass in diabetes by changing inflammatory cytokines and stress factors such as Hsp70 in skeletal muscle and adipose tissue.     

Effects of low-load resistance training with vascular occlusion on the mechanical properties of muscle and tendon

The present study aimed to investigate the effects of low-load resistance training with vascular occlusion on the specific tension and tendon properties by comparing with those of high-load training. Nine participants completed 12 weeks (3 days/week) of a unilateral isotonic training program on knee extensors. One leg was trained using low load (20% of 1 RM) with vascular occlusion (LLO) and other leg using high load (80% of 1 RM) without vascular occlusion (HL). Before and after training, maximal isometric knee extension torque (MVC) and muscle volume were measured. Specific tension of vastus lateralis muscle (VL) was calculated from MVC, muscle volume, and muscle architecture measurements. Stiffness of tendon-aponeurosis complex in VL was measured using ultrasonography during isometric knee extension. Both protocols significantly increased MVC and muscle volume of quadriceps femoris muscle. Specific tension of VL increased significantly 5.5% for HL, but not for LLO. The LLO protocol did not alter the stiffness of tendon-aponeurosis complex in knee extensors, while the HL protocol increased it significantly. The present study demonstrated that the specific tension and tendon properties were found to remain following low-load resistance training with vascular occlusion, whereas they increased significantly after high-load training.     

抗阻训练通过Akt-FoxO1通路抑制低氧诱导的骨骼肌萎缩

高原低氧环境会引起肌力下降和运动能力退化,而抗阻训练是刺激骨骼肌生长的重要手段,叉头转录因子1（fork head box protein O 1,FoxO1）在调控骨骼肌蛋白质分解通路中承担重要角色。为探究Akt-FoxO1通路是否参与抗阻训练抑制低氧诱导的骨骼肌萎缩,本研究构建低氧诱导骨骼肌萎缩的大鼠模型,并模拟海拔4 000 m低氧环境下（12.4% O2）进行抗阻训练,对比观察大鼠比目鱼肌和趾长伸肌湿重和横截面积,以及蛋白激酶B（protein kinase B,Akt）、叉头转录因子1、泛素蛋白连接酶1（muscle ring finger 1,MuRF1）的表达差异等。结果表明,低氧暴露导致大鼠趾长伸肌湿重显著下降,苏木精-伊红染色组织切片分析肌纤维横截面积、低氧环境下比目鱼肌横截面积明显下降,而低氧抗阻训练后趾长伸肌横截面积明显高于安静组。实时荧光定量PCR和蛋白质免疫印迹结果显示,低氧暴露后FoxO1和MuRF1基因表达明显上调,低氧下抗阻训练后发现,Akt基因表达明显上调而FoxO1、MuRF则明显下调。免疫荧光观察磷酸化FoxO1在细胞核内外表达情况,发现抗阻训练后FoxO1（S256）于细胞核外表达增强。上述结果表明,抗阻训练可以达到抑制低氧诱导骨骼肌萎缩的效果,Akt促进FoxO1磷酸化从而减缓骨骼肌蛋白质分解过程是抗阻训练能够抑制骨骼肌萎缩的分子机制之一。     

Effect of prolonged intermittent hypoxia and exercise training on glucose tolerance and muscle GLUT4 protein expression in rats

We compared the chronic effect of intermittent hypoxia and endurance training on the glucose tolerance and GLUT4 protein expression in rat skeletal muscle. Thirty-two Sprague-Dawley rats were matched for weight and assigned to one of the following four groups: control, endurance training, hypoxia, or hypoxia followed by endurance training. Hypoxic treatment consisted of breathing 14% O₂ for 12 h/day under normobaric conditions, and the training protocol consisted of making animals swim 2 times for 3 h/day. At the end of the 3rd week, an oral glucose tolerance test (OGTT) was performed 16 h after treatments. At the end of the 4th week, GLUT4 protein, mRNA, and glycogen storage in skeletal muscle were determined. Endurance training significantly improved OGTT results. Glycogen content and GLUT4 protein expression in the plantaris and red gastrocnemius, but not in the soleus or white gastrocnemius muscles, were also elevated. Chronic intermittent hypoxia also improved OGTT results, but did not alter GLUT4 protein expression. Additionally, hypoxia followed by exercise training produced significant increases in GLUT4 protein and mRNA in a greater number of muscles compared to endurance training alone. Both exercise training and hypoxia significantly reduced body mass, and an additive effect of both treatments was found. In conclusion, chronic intermittent hypoxia improved glucose tolerance in the absence of increased GLUT4 protein expression. This treatment facilitated the exercise training effect on muscle GLUT4 expression and glycogen storage. These new findings open the possibility of utilizing intermittent hypoxia, with or without exercise training, for the prevention and clinical treatment of type 2 diabetes or insulin resistance.     

Effect of creatine supplementation during high resistance training on mass,strength, and fatigue resistance in rat skeletal muscle

The aim of this study was to utilize a rodent model of resistance exercise to compare training with creatine supplementation with training alone. We tested the hypothesis that creatine supplementation during high resistance training would result in greater increases in muscle mass, contractile force, and superior resistance to fatigue compared with training alone. Two groups of rats underwent training of the tibialis anterior muscle (TA) for 4 weeks without creatine (NCr group) or with creatine (0.5 g.kg(-1).d(-1)) (CrT group). The relative loads in each animal were held constant during the training protocol. Training resulted in comparable significant increases in muscle contractile force in both the NCr and CrT groups. Creatine supplementation did not result in a significant increase in fatigue resistance and resulted in a significant decrease in postfatigue recovery compared with training alone. Training resulted in a significant increase in muscle dry weight in both groups, whereas muscle wet weight gains in the CrT group were double the gains in the NCr group. The data from this study suggest that for creatine to have a beneficial effect on muscle strength and mass beyond training alone, the workloads need to be adjusted. That is, any potential benefit of creatine to enable a greater lifting volume during resistance training needs to be incorporated into the training regime for creatine to be effective.     

Effects of aging on vasoconstrictor and mechanical properties of rat skeletal muscle arterioles

Exercise capacity and skeletal muscle blood flow during exercise are reduced with advancing age. This reduction in blood flow capacity may be related to increased reactivity of skeletal muscle resistance vessels to vasoconstrictor stimuli. The purpose of this study was to test the hypothesis that aging results in increased vasoconstrictor responses of skeletal muscle resistance arterioles. First-order (1A) arterioles (90-220 microm) from the gastrocnemius and soleus muscles of young (4 mo) and aged (24 mo) Fischer-344 rats were isolated, cannulated, and pressurized via hydrostatic reservoirs. Vasoconstriction in response to increases in norepinephrine (NE; 1 x 10(-9)-1 x 10(-4) M) and KCl (20-100 mM) concentrations and increases in intraluminal pressure (10-130 cmH(2)O) were evaluated in the absence of flow. Responses to NE and KCl were similar in both soleus and gastrocnemius muscle arterioles from young and aged rats. In contrast, active myogenic responses to changes in intraluminal pressure were diminished in soleus and gastrocnemius arterioles from aged rats. To assess whether alterations in the mechanical properties of resistance arterioles underlie altered myogenic responsiveness, passive diameter responses to pressure and mechanical stiffness were evaluated. There was no effect of age on the structural behavior (passive pressure-diameter relationship) or stiffness of arterioles from either the soleus or gastrocnemius muscles. These results suggest that aging does not result in a nonspecific decrease in vasoconstrictor responsiveness of skeletal muscle arterioles. Rather, aging-induced adaptations of vasoreactivity of resistance arterioles appear to be limited to mechanisms that are uniquely involved in the signaling of the myogenic response.     

Effects of intensive physical rehabilitation on neuromuscular adaptations in adults with poststroke hemiparesis

Hemiparesis-disability and muscle weakness of 1 side of the body-is a common consequence of stroke. High-intensity strength training may be beneficial to regain function, but strength coaches in the field of rehabilitation need evidence-based guidelines. The purpose of this study was to evaluate the effect of intensive physical rehabilitation on neuromuscular and functional adaptations in outpatients suffering from hemiparesis after stroke. A within-subject repeated-measures design with the paretic leg as the experimental leg and the nonparetic leg as the control leg was used. Eleven outpatients with hemiparesis after stroke participated in 12 weeks of intensive physical rehabilitation comprising unilateral high-intensity strength training with near-maximal loads (4-12 repetition maximum) and body weight supported treadmill training. At baseline and 12-week follow-up, the patients went through testing consisting of isokinetic muscle strength, neuromuscular activation measured with electromyography (EMG), electrically evoked muscle twitch contractile properties, and gait performance (10-m Walk Test and 6-min Walk Test). After the 12-week conditioning program, knee extensor and flexor strength increased during all contraction modes and velocities in the paretic leg. Significant increases were observed for agonist EMG amplitude at slow concentric and slow eccentric contraction. Twitch torque increased, whereas twitch time-to-peak tension remained unchanged. By contrast, no significant changes were observed in the nonparetic control leg. Gait performance increased 52-68%. In conclusion, intensive physical rehabilitation after stroke leads to clinically relevant neuromuscular improvements, leading to increased voluntary strength during a wide range of contraction modes and velocities, and improved gait velocity. Strength training coaches working in the field of rehabilitation can use this knowledge to safely and efficiently add high-intensity strength training to existing rehabilitation paradigms.     

Metformin, but not exercise training, increases insulin responsiveness in skeletal muscle of Sprague-Dawley rats

We assessed the effects of combined metformin treatment and exercise training on body composition, on insulin concentration following glucose loading, on insulin-stimulated glucose transport in skeletal muscle, and on muscle glycogen content. Male Sprague-Dawley rats were treated for 35 days with or without metformin (320 mg/kg/day) and/or treadmill exercise training (20 min at 20 m/min, 5 days/wk). Because metformin reduces food intake, pair-fed controls were included. Metformin, training, and pair-feeding all decreased food intake, body weight, and insulin concentration following glucose loading. Metformin and training reduced intra-abdominal fat, but pair feeding did not. In isolated strips derived from soleus, epitrochlearis and extensor carpi ulnaris muscles, metformin increased insulin-stimulated transport of [3H]-2-deoxyglucose by 90%, 89% and 125%, respectively (P < 0.02) and training increased [3H]-2-deoxyglucose transport in the extensor carpi ulnaris muscle only (66%, P < 0.05). Pair-feeding did not alter [3H]-2-deoxyglucose transport. Training increased gastrocnemius muscle glycogen by 100% (P < 0.001). Metformin and pair-feeding did not alter muscle glycogen. We conclude that metformin reverses the maturation-induced impairment of insulin responsiveness in Sprague-Dawley rats by increasing insulin-stimulated glucose transport in skeletal muscle and that this effect is not secondary to reduced food intake. We also conclude that metformin and exercise training may increase insulin sensitivity by different mechanisms, with training causing increased glucose transport only in some muscles and also causing increased muscle glycogen storage.     

Antioxidant enzyme activity is up-regulated after unilateral resistance exercise training in older adults

Cellular antioxidant capacity and oxidative stress are postulated to be critical factors in the aging process. The effects of resistance exercise training on the level of skeletal muscle oxidative stress and antioxidant capacity have not previously been examined in older adults. Muscle biopsies from both legs were obtained from the vastus lateralis muscle of 12 men 71 +/- 7 years of age. Subjects then engaged in a progressive resistance exercise-training program with only one leg for 12 weeks. After 12 weeks, the nontraining leg underwent an acute bout of exercise (exercise session identical to that of the trained leg at the same relative intensity) at the same time as the last bout of exercise in the training leg. Muscle biopsies were collected from the vastus lateralis of both legs 48 h after the final exercise bout. Electron transport chain enzyme activity was unaffected by resistance training and acute resistance exercise (p < 0.05). Training resulted in a significant increase in CuZnSOD (pre--7.2 +/- 4.2, post--12.6 +/- 5.6 U.mg protein(-1); p = 0.02) and catalase (pre--8.2 +/- 2.3, post--14.9 +/- 7.6 micromol.min(-1).mg protein(-1); p = 0.02) but not MnSOD activity, whereas acute exercise had no effect on the aforementioned antioxidant enzyme activities. Furthermore, basal muscle total protein carbonyl content did not change as a result of exercise training or acute exercise. In conclusion, unilateral resistance exercise training is effective in enhancing the skeletal muscle cellular antioxidant capacity in older adults. The potential long-term benefits of these adaptations remain to be evaluated.     

Testosterone stimulates myoglobin expression in different muscles of the mouse

The regulation of energy metabolism is one of the major functions of steroid hormones. This study was performed to explore whether testosterone can regulate the aerobic capacity of skeletal muscles via myoglobin expression. To study this, changes in testosterone level were quantified, and the level of myoglobin protein was analyzed using Western blot in mice subjected to 6 weeks of training (T) or testosterone administration (A). Both treatments significantly increased the plasma testosterone level when compared to the untrained (U) or control (C) group. Training induced a significant increase in the myoglobin content in gastrocnemius and plantaris muscles (287 and 83%, respectively). Testosterone administration increased myoglobin concentration in plantaris (183%) but not in gastrocnemius. In extensor digitorum longus muscle the protein content decreased slightly after exercise, but increased 78% after testosterone administration. In soleus and rectus femoris muscles the myoglobin content was unchanged after both treatments. The data show that testosterone and training have differential effects on the concentration of myoglobin in some, but not all muscles. This may have an influence on the aerobic capacity in mouse skeletal muscles. The data demonstrated that both testosterone administration and training induced an increase in plasma testosterone level. However, the effects of the treatments on the myoglobin concentration differ.     

Hindlimb casting decreases muscle mass in part by proteasome-dependent proteolysis but independent of protein synthesis

The hypothesis of the present study was that rats subjected to short-term unilateral hindlimb immobilization would incur skeletal muscle wasting and concomitant alterations in protein synthesis, controllers of translation, and indexes of protein degradation. Rats were unilaterally casted for 1, 3, or 5 days to avoid complications associated with other disuse models. In the casted limb, gastrocnemius wet weight decreased 12% after 3 days and thereafter remained constant. In contrast, the contralateral control leg displayed a steady growth rate over time. The rate of protein synthesis and translational efficiency were unchanged in the immobilized muscle at day 5. The total amount and phosphorylation state of regulators of translational initiation and elongation were unaltered. The mRNA contents of polyubiquitin and the ubiquitin ligases muscle atrophy F-box (MAFbx)/Atrogin-1 and muscle RING finger 1 (MuRF1) were elevated in immobilized muscle at all time points, with peak expression occurring at day 3. Daily injection of the type II glucocorticoid receptor antagonist RU-486 did not prevent decreases in gastrocnemius wet weight nor increases in mRNA for MAFbx/Atrogin-1 and MuRF1. However, in vivo administration of the proteasome inhibitor Velcade prevented 53% of wet weight loss associated with 3 days of immobilization. These data suggest that the loss of skeletal muscle mass in this model of disuse appears to be glucocorticoid independent, can be partially rescued with a potent proteasome inhibitor, and is associated with enhanced mRNA expression of multiple factors that contribute to ubiquitin- proteasome-dependent degradation and are likely to control the remodeling of immobilized skeletal muscle during atrophy.     

Skeletal muscle and liver glutathione homeostasis in response to training, exercise, and immobilization.

Female beagle dogs were treadmill trained 40 km/day at 5.5-6.8 km/h, 15% upgrade, 5 days/wk for 55 wk. With training, hepatic and red gastrocnemius (RG) total glutathione increased, glutathione peroxidase (GPX) and glutathione reductase (GRD) increased in all the leg muscles studied, and hepatic glutathione S-transferase (GST) activity increased. Joint immobilization (11 wk) did not affect GPX, GRD, and GST of RG, but total glutathione decreased. Male Han Wistar rats were treadmill trained 2 h/day at 2.1 km/h, 5 days/wk for 8 wk. With training, hepatic total glutathione and leg muscle GPX increased but GRD of RG decreased, perhaps because of an increased muscle flavo-protein breakdown during exhaustive training. gamma-Glutamyl transpeptidase was higher in the trained leg muscles. Exhaustive exercise decreased muscle gamma-glutamyl transpeptidase of only control leg muscle, depleted muscle (lesser extent in trained rats) and liver total glutathione of both groups, decreased GRD only in untrained RG, and increased hepatic GST. Endurance training elevated the antioxidant and detoxicant status of muscle and liver, respectively.     

Exercise training blunts microvascular rarefaction in the metabolic syndrome

Reduced skeletal muscle microvessel density (MVD) in the obese Zucker rat (OZR) model of the metabolic syndrome is a function of a chronic reduction in vascular nitric oxide (NO) bioavailability. Previous studies suggest that exercise can improve NO bioavailability and reduce chronic inflammation and that low vascular NO bioavailability may be associated with impaired angiogenic responses via increased matrix metalloproteinase (MMP)-2 and MMP-9 activity. As such, we hypothesized that chronic exercise (EX) would increase NO bioavailability in OZR and blunt microvascular rarefaction through reduced MMP activity, and potentially via altered plasma cytokine levels. Ten weeks of treadmill exercise (1 h/day, 5 days/wk, 22 m/min) reduced body mass and fasting insulin and triglyceride levels in EX-OZR vs. sedentary (SED) OZR. In EX-OZR, gastrocnemius muscle MVD was improved by 19 +/- 4%, whereas skeletal muscle arteriolar dilation and conduit arterial methacholine-induced NO release were increased. In EX-OZR, functional hyperemia was improved vs. SED-OZR, and minimum vascular resistance within perfused gastrocnemius muscle was reduced, although no change in arteriolar stiffness was identified. Western blotting and gelatin zymography demonstrated that neither expression nor activity of MMP-2 or MMP-9 was altered in skeletal muscle of EX vs. SED animals. Plasma markers of inflammation associated with angiogenesis, monocyte chemoattractant protein-1 and IL-1beta, were increased in SED-OZR and were reduced with training, whereas IL-13 was reduced in SED-OZR and increased with exercise. These data suggest that exercise-induced improvements in skeletal muscle MVD in OZR are associated with increased NO bioavailability and may stem from altered inflammatory profiles rather than MMP function.