Local hindlimb antioxidant infusion does not affect muscle glucose uptake during in situ contractions in rat

Local hindlimb antioxidant infusion does not affect muscle glucose uptake during in situ contractions in rat. J Appl Physiol 108: 1275-1283, 2010. First published March 4, 2010; doi:10.1152/japplphysiol.01335.2009.-There is evidence that reactive oxygen species (ROS) contribute to the regulation of skeletal muscle glucose uptake during highly fatiguing ex vivo contraction conditions via AMP-activated protein kinase (AMPK). In this study we investigated the role of ROS in the regulation of glucose uptake and AMPK signaling during low-moderate intensity in situ hindlimb muscle contractions in rats, which is a more physiological protocol and preparation. Male hooded Wistar rats were anesthetized, and then N-acetylcysteine (NAC) was infused into the epigastric artery (125 mgkg1h1) of one hindlimb (contracted leg) for 15 min before this leg was electrically stimulated (0.1-ms impulse at 2 Hz and 35 V) to contract at a low-moderate intensity for 15 min. The contralateral leg did not receive stimulation or local NAC infusion (rest leg). NAC infusion increased (P0.05) plasma cysteine and cystine (by 360- and 1.4-fold, respectively) and muscle cysteine (by 1.5-fold, P0.001). Although contraction did not significantly alter muscle tyrosine nitration, reduced (GSH) or oxidized glutathione (GSSG) content, S-glutathionylation of protein bands at 250 and 150 kDa was increased (P0.05) 1.7-fold by contraction, and this increase was prevented by NAC. Contraction increased (P0.05) skeletal muscle glucose uptake 20-fold, AMPK phosphorylation 6-fold, ACC| phosphorylation 10-fold, and p38 MAPK phosphorylation 60-fold, and the muscle fatigued by 30% during contraction and NAC infusion had no significant effect on any of these responses. This was despite NAC preventing increases in S-glutathionylation with contraction. In conclusion, unlike during highly fatiguing ex vivo contractions, local NAC infusion during in situ low-moderate intensity hindlimb contractions in rats, a more physiological preparation, does not attenuate increases in skeletal muscle glucose uptake or AMPK signaling.