Skeletal muscle signaling, metabolism, and performance during sprint exercise in severe acute hypoxia after the ingestion of antioxidants

The aim of this study was to determine if reactive oxygen species (ROS) could play a role in blunting Thr(172)-AMP-activated protein kinase (AMPK)alpha-phosphorylation in human skeletal muscle after sprint exercise in hypoxia and to elucidate the potential signaling mechanisms responsible for this response. Nine volunteers performed a single 30-s sprint (Wingate test) in two occasions while breathing hypoxic gas (PIO2 = 75 mmHg): one after the ingestion of placebo and another following the intake of antioxidants (alpha-lipoic acid, vitamin C, and vitamin E), with a randomized double-blind design. Vastus lateralis muscle biopsies were obtained before, immediately after, and 30- and 120-min post-sprint. Compared with the control condition, the ingestion of antioxidants resulted in lower plasma carbonylated proteins, attenuated elevation of the AMP-to-ATP molar ratio, and reduced glycolytic rate (P < 0.05) without significant effects on performance or V-O2. The ingestion of antioxidants did not alter the basal muscle signaling. Thr(172)-AMPK alpha and Thr(184/187)- transforming growth factor-beta-activated kinase 1 (TAK1) phosphorylation were not increased after the sprint regardless of the ingestion of antioxidants. Thr(286)-CaMKII phosphorylation was increased after the sprint, but this response was blunted by the antioxidants. Ser(485)-AMPK-1/Ser(491)-AMPK alpha 2 phosphorylation increased immediately after the sprints coincident with increased Akt phosphorylation. In summary, antioxidants attenuate the glycolytic response to sprint exercise in severe acute hypoxia and modify the muscle signaling response to exercise. Ser(485)-AMPK alpha 1/Ser(491)-AMPK alpha 2 phosphorylation, a known mechanism of Thr(172)-AMPK alpha phosphorylation inhibition, is increased immediately after sprint exercise in hypoxia, probably by a mechanism independent of ROS. NEW & NOTEWORTHY The glycolytic rate is increased during sprint exercise in severe acute hypoxia. This study showed that the ingestion of antioxidants before sprint exercise in severe acute hypoxia reduced the glycolytic rate and attenuated the increases of the AMP-to-ATP and the reduction of the NAD(+) -to-NADH.H+ ratios. This resulted in a modified muscle signaling response with a blunted Thr(286)-CaMKII but similar AMP-activated protein kinase phosphorylation responses in the sprints preceded by the ingestion of antioxidants.