Pre-heating stress associated with acute oral leucine supplementation effects in rat gastrocnemius muscle: Implications for protein synthesis signaling pathways.

Skeletal muscle is a highly plastic tissue. The role of heat shock protein 72 (Hsp72) in heat stress-induced skeletal muscle hypertrophy has been well demonstrated; however, the precise mechanisms remain unclear. Essential amino acids, such as leucine, mainly mediate muscle protein synthesis. We investigated the effects of pre-heating and increased Hsp72 expression on the mechanistic target of rapamycin (mTOR) signaling and protein synthesis following leucine administration in rat gastrocnemius muscle. To ensure increased Hsp72 expression in both the red and white portions of the muscle, one leg of male Wistar rats (10-week-old, n = 23) was heat-stressed in 43 °C water for 30 min twice at a 48-h-interval (heat-stressed leg, HS leg). The contralateral leg served as a non-heated internal control (CT leg). After the recovery period (48 h), rats were divided into the pre-administration or oral leucine administration groups. We harvested the gastrocnemius muscle (red and white parts) prior to administration and 30 and 90 min after leucine treatment (n = 7-8 per group) and intramuscular signaling responses to leucine ingestion were determined using western blotting. Heat stress significantly upregulated the expression of Hsp72 and was not altered by leucine administration. Although the phosphorylation levels of mTOR/S6K1 and ERK were similar regardless of heating, 4E-BP1 was less phosphorylated in the HS legs than the CT legs after leucine administration in the red portion of the muscles (P < 0.05). Moreover, c-Myc expression differed significantly after leucine administration in both the red and white portions of the muscles. Our findings indicate that following oral leucine administration, pre-heating partially blunted the muscle protein synthesis signaling response in the rat gastrocnemius muscle.

Effect of a combination of astaxanthin supplementation, heat stress, and intermittent reloading on satellite cells during disuse muscle atrophy.

We examined the effect of a combination of astaxanthin (AX) supplementation, repeated heat stress, and intermittent reloading (IR) on satellite cells in unloaded rat soleus muscles. Forty-nine male Wistar rats (8-week-old) were divided into control, hind-limb unweighting (HU), IR during HU, IR with AX supplementation, IR with repeated heat stress (41.0-41.5 °C for 30 min), and IR with AX supplementation and repeated heat stress groups. After the experimental period, the antigravitational soleus muscle was analyzed using an immunohistochemical technique. Our results revealed that the combination of dietary AX supplementation and heat stress resulted in protection against disuse muscle atrophy in the soleus muscle. This protective effect may be partially due to a higher satellite cell number in the atrophied soleus muscle in the IR/AX/heat stress group compared with the numbers found in the other groups. We concluded that the combination treatment with dietary AX supplementation and repeated heat stress attenuates soleus muscle atrophy, in part by increasing the number of satellite cells.

Dietary astaxanthin supplementation attenuates disuse-induced muscle atrophy and myonuclear apoptosis in the rat soleus muscle.

Extended periods of skeletal muscle disuse results in muscle atrophy and weakness. Currently, no therapeutic treatment is available for the prevention of this problem. Nonetheless, growing evidence suggests that prevention of disuse-induced oxidative stress in inactive muscle fibers can delay inactivity-induced muscle wasting. Therefore, this study tested the hypothesis that dietary supplementation with the antioxidant astaxanthin would protect against disuse muscle atrophy, in part, by prevention of myonuclear apoptosis. Wistar rats (8 weeks old) were divided into control (CT, n = 9), hindlimb unloading (HU, n = 9), and hindlimb unloading with astaxanthin (HU + AX, n = 9) groups. Following 2 weeks of dietary supplementation, rats in the HU and HU + AX groups were exposed to unloading for 7 days. Seven-day unloading resulted in reduced soleus muscle weight and myofiber cross-sectional area (CSA) by ~30 and ~47 %, respectively. Nonetheless, relative muscle weights and CSA of the soleus muscle in the HU + AX group were significantly greater than those of the HU group. Moreover, astaxanthin prevented disuse-induced increase in the number of terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL)-positive nuclei. We conclude that astaxanthin supplementation prior to and during hindlimb unloading attenuates soleus muscle atrophy, in part, by suppressing myonuclear apoptosis.

Astaxanthin intake attenuates muscle atrophy caused by immobilization in rats.

Astaxanthin is a carotenoid pigment and has been shown to be an effective inhibitor of oxidative damage. We tested the hypothesis that astaxanthin intake would attenuate immobilization-induced muscle atrophy in rats. Male Wistar rats (14-week old) were fed for 24 days with either astaxanthin or placebo diet. After 14 days of each experimental diet intake, the hindlimb muscles of one leg were immobilized in plantar flexion position using a plaster cast. Following 10 days of immobilization, both the atrophic and the contralateral plantaris muscles were removed and analyzed to determine the level of muscle atrophy along with measurement of the protein levels of CuZn-superoxide dismutase (CuZn-SOD) and selected proteases. Compared with placebo diet animals, the degree of muscle atrophy in response to immobilization was significantly reduced in astaxanthin diet animals. Further, astaxanthin supplementation significantly prevented the immobilization-induced increase in the expression of CuZn-SOD, cathepsin L, calpain, and ubiquitin in the atrophied muscle. These results support the postulate that dietary astaxanthin intake attenuates the rate of disuse muscle atrophy by inhibiting oxidative stress and proteolysis via three major proteolytic pathways.

Effects of Electrostimulation with Blood Flow Restriction on Muscle Size and Strength.

PURPOSE: Low-load voluntary exercise can induce muscle hypertrophy and strength gain in working muscles when combined with blood flow restriction (BFR). However, it is unknown whether such hypertrophy and strength gain can be induced by involuntary muscle contractions triggered via low-intensity neuromuscular electrical stimulation (NMES) combined with BFR. The purpose of this article was to investigate whether low-intensity NMES combined with BFR (NMES-BFR) could elicit muscle hypertrophy and strength gain in the quadriceps. METHODS: Eight untrained young male participants (mean ± SE; age, 26.2 ± 0.7 yr; height, 1.74 ± 0.02 m; body weight, 71.4 ± 4.8 kg) were subjected to 23 min of unilateral low-intensity (5%-10% of maximal voluntary contraction) NMES twice per day (5 d·wk⁻¹) for 2 wk: one leg received NMES-BFR and the other leg received NMES alone. Quadriceps muscle thickness and isometric and isokinetic strength were measured before and every week throughout the training and detraining periods. RESULTS: In NMES-BFR legs, muscle thickness increased after 2 wk of training (+3.9%) and decreased after 2 wk of detraining (-3.0%). NMES-BFR training also increased maximal knee extension strength in isometric (+14.2%) and isokinetic (+7.0% at 90°·s⁻¹ and +8.3% at 180°·s⁻¹) voluntary contractions. In addition, maximal isometric strength decreased (-6.8%), whereas no large fall (-1.9% at 90°·s⁻¹ and -0.6% at 180°·s⁻¹) in isokinetic maximal strength was evident after 2 wk of detraining. In legs that received NMES alone, no prominent change was observed; there was a negligible effect on isometric strength. CONCLUSION: Low-intensity NMES-BFR induces muscle hypertrophy and strength gain in untrained young male participants.

Microwave hyperthermia treatment increases heat shock proteins in human skeletal muscle.

OBJECTIVE: To test the hypothesis that microwave hyperthermia treatment (MHT) increases heat shock proteins (HSPs) in the human vastus lateralis muscle. METHODS: Four untrained healthy male volunteers participated in this study. The lateral side of the thigh of one leg (heated leg) was heated with a microwave generator (2.5 GHz, 150 W) for 20 min. At 1 day after the MHT, a muscle sample was taken from the heated leg. A control sample was taken from the unheated leg on another day of the MHT. For both legs, HSP90, HSP72 and HSP27 levels were compared. RESULTS: The HSP90, HSP72 and HSP27 levels in heated legs were significantly higher than those in control legs (p<0.05). CONCLUSIONS: Application of MHT can increase the levels of several HSPs in human skeletal muscle.

Changes in muscle temperature induced by 434 MHz microwave hyperthermia.

OBJECTIVE: To investigate the changes in temperature of human muscle during microwave hyperthermia. METHODS: Skin surface and muscle temperatures were measured in 11 healthy adult men (mean (SD) age 24.3 (2.2) years; height 174.2 (6.1) cm; weight 70.0 (5.3) kg) during a 30 min exposure of the thigh to 434 MHz microwave hyperthermia. Skin temperature was maintained at the pilot temperature of 40 degrees C, and the temperature of the water in the bolus was 38 degrees C. The peak power output was set at 60 W and controlled automatically to maintain the pilot temperature. The temperature was measured in the vastus lateralis muscle at an average muscle depth of 2.0 (0.2) cm, using a 23 G Teflon-shielded thermocouple. Biopsy specimens were obtained for light microscopy from three subjects. A muscle-equivalent phantom was used to evaluate the vertical heating pattern. RESULTS: Both skin and muscle temperatures increased from baseline, and muscle temperature was higher than skin temperature (skin temperature 39.2 (0.5) degrees C, temperature rise 5.0 (1.5) degrees C; muscle temperature 43.7 (0.8) degrees C, temperature rise 8.9 (1.4) degrees C). At the end of the hyperthermia treatment, muscle temperature decreased to 39.8 (0.9) degrees C, but was still 4.8 (1.5) degrees C higher than the baseline. No signs of muscle damage were observed on the basis of the blood creatine kinase activity and histological sections. CONCLUSIONS: The results show that the 434 MHz microwave hyperthermia treatment increased and maintained muscle temperature locally by 6.3-11.4 degrees C without muscle damage. These findings suggest that the microwave hyperthermia system provides effective and safe treatment.