Skeletal muscle autophagy remains responsive to hyperinsulinemia and hyperglycemia at higher plasma insulin concentrations in insulin-resistant mice.

Skeletal muscle autophagy is suppressed by insulin, but it is not clear if such suppression is altered with insulin resistance. We investigated if the inhibitory action of insulin on autophagy remains intact despite insulin resistance to glucose metabolism. C57BL/6J mice consumed either a low-fat (10% fat) diet as control or high-fat (60% fat) diet for 12 weeks to induce insulin resistance. Following a 5-hour fast, mice underwent either hyperinsulinemic-euglycemic, hyperinsulinemic-hyperglycemic, or saline infusion to test the effect of insulin on autophagy markers in the quadriceps muscle (n = 8-10 per diet and clamp condition). Mice were anesthetized by sodium pentobarbital for tissue collection after 2 h of infusion. Despite the high-fat group having lower insulin-stimulated glucose uptake, both low-fat and high-fat groups had similar autophagosome abundance during hyperinsulinemic conditions. The lipidation of microtubule-associated proteins 1A/1B light chain 3B (LC3II/LC3I) was decreased in hyperinsulinemia versus saline control (P < 0.01) in low-fat (-54%) and high-fat groups (-47%), demonstrating similar suppression of autophagy between diet groups. Mitochondrial-associated LC3II was greater in the high-fat compared to the low-fat group (P = 0.045) across clamp conditions, suggesting a greater localization of autophagosomes with mitochondria. L6 myotubes were treated with insulin and rapamycin to determine the role of mechanistic target of rapamycin complex-1 (mTORC1) in insulin-mediated suppression of autophagy. Inhibition of mTORC1 blunted the decline of LC3II/LC3I with insulin by 40%, suggesting mTORC1 partially mediates the insulin action to suppress autophagy. Collectively, autophagy remained responsive to the suppressive effects of insulin in otherwise insulin-resistant and obese mice.