In skeletal muscle cells, GLUT1 is responsible for a large portion of basal uptake of glucose and dehydroascorbic acid, both of which play roles in antioxidant defense. We hypothesized that conditions that would decrease GLUT1-mediated transport would cause increased reactive oxygen species (ROS) levels in L6 myoblasts, while conditions that would increase GLUT1-mediated transport would result in decreased ROS levels. We found that the GLUT1 inhibitors fasentin and phloretin increased the ROS levels induced by antimycin A and the superoxide generator pyrogallol. However, indinavir, which inhibits GLUT4 but not GLUT1, had no effect on ROS levels. Ataxia telangiectasia mutated (ATM) inhibitors and activators, previously shown to inhibit and augment GLUT1-mediated transport, increased and decreased ROS levels, respectively. Mutation of an ATM target site on GLUT1 (GLUT1-S490A) increased ROS levels and prevented the ROS-lowering effect of the ATM activator doxorubicin. In contrast, expression of GLUT1-S490D lowered ROS levels during challenge with pyrogallol, prevented an increase in ROS when ATM was inhibited, and prevented the pyrogallol-induced decrease in insulin signaling and insulin-stimulated glucose transport. Taken together, the data suggest that GLUT1 plays a role in regulation of ROS and could contribute to maintenance of insulin action in the presence of ROS.
Elevated reactive oxygen species (ROS) levels are associated with metabolic abnormalities such as insulin resistance. Glucose transporter 1 (GLUT1) transports glucose and dehydroascorbic acid, both of which play roles in antioxidant defense. We hypothesized that conditions that would increase GLUT1 activity would result in decreased ROS levels and conditions that would decrease GLUT1 activity would increase ROS levels. . L6 myoblasts loaded with H2‐DCFDA, a fluorescent ROS probe, were treated under various conditions to alter GLUT1 activity, and fluorescence readings were taken under basal conditions or after increases in ROS were induced by exposure to pyrogallol or antimycin A. GLUT1 inhibitors, fasentin and phloretin, increased induced ROS levels. Ataxia telangiectasia mutated inhibitors and activators, previously shown to inhibit and augment GLUT1 activity, increased and decreased basal and induced ROS levels, respectively. GLUT1‐S490A, previously shown to decrease GLUT1 activity, enhanced basal and induced ROS levels. GLUT1‐S490D, previously shown to increase GLUT1‐mediated transport, lowered basal ROS levels. Thus, the data suggest that GLUT1 plays a role in regulation of ROS levels.
Grant Funding Source: Supported by: William Townsend Porter Predoctoral Fellowship from the American Physiological Society
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