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.

Impact of Dietary Nitrate Supplementation on Executive Function During Hypoxic Exercise.

We investigated the effects of 4-day dietary beetroot (BR) juice supplementation on executive function assessed by color-word Stroop task (CWST) in hypoxia (fraction of inspired oxygen [FiO2] = 0.1395). Eight healthy young men performed 25-minute leg cycling exercise (target heart rate, 140 bpm) randomly with placebo (PL) juice or BR supplementation. The CWST was evaluated at resting hypoxic condition (after 30 minutes of hypoxic exposure) and at 15 minutes during exercise. During exercise, the correct response time in the CWST was significantly shortened compared with those at rest with no differences between PL and BR. The response accuracy in the CWST with BR was marginally lower than that with PL during exercise (p = 0.066). There were no significant differences in all physiological values, including pulmonary ventilation, arterial oxygen saturation, partial pressure of end-tidal carbon dioxide output, and tissue oxygenation in the left frontal lobe, assessed by near-infrared spectroscopy during exercise conditions involving PL and BR supplementation. These results suggest that moderate exercise in hypoxia partially improved executive function; however, 4-day dietary BR supplementation did not improve executive function during hypoxic exercise.

Muscle oxygenation profiles between active and inactive muscles with nitrate supplementation under hypoxic exercise.

Whether dietary nitrate supplementation improves exercise performance or not is still controversial. While redistribution of sufficient oxygen from inactive to active muscles is essential for optimal exercise performance, no study investigated the effects of nitrate supplementation on muscle oxygenation profiles between active and inactive muscles. Nine healthy males performed 25 min of submaximal (heart rate ~140 bpm; EXsub) and incremental cycling (EXmax) until exhaustion under three conditions: (A) normoxia without drink; (B) hypoxia (FiO2 = 13.95%) with placebo (PL); and (c) hypoxia with beetroot juice (BR). PL and BR were provided for 4 days. Oxygenated and deoxygenated hemoglobin (HbO2 and HHb) were measured in vastus lateralis (active) and biceps brachii (inactive) muscles, and the oxygen saturation of skeletal muscle (StO2; HbO2/total Hb) were calculated. During EXsub, BR suppressed the HHb increases in active muscles during the last 5 min of exercise. During EXmax, time to exhaustion with BR (513 ± 24 sec) was significantly longer than with PL (490 ± 39 sec, P < 0.05). In active muscles, BR suppressed the HHb increases at moderate work rates during EXmax compared to PL (P < 0.05). In addition, BR supplementation was associated with greater reductions in HbO2 and StO2 at higher work rates in inactive muscles during EXmax Collectively, these findings indicate that short-term dietary nitrate supplementation improved hypoxic exercise tolerance, perhaps, due to suppressed increases in HHb in active muscles at moderate work rates. Moreover, nitrate supplementation caused greater reductions in oxygenation in inactive muscle at higher work rates during hypoxic exercise.