This study aimed to test whether stimulation of net hepatic glucose output (NHGO) by increased concentrations of the adenosine 5'-monophosphate analogue, 5-aminoimidazole-4-carboxamide-1--D-ribosyl-5-monophospate, can be suppressed by pharmacological insulin levels. Dogs had sampling (artery, portal vein, hepatic vein) and infusion (vena cava, portal vein) catheters, and flow probes (hepatic artery, portal vein) implanted >16 days prior to a study. Protocols consisted of equilibration (-130 to -30 min), basal (-30 to 0 min) and hyperinsulinemic-euglycemic (0 to 150 min) periods. At t=0 min, somatostatin was infused and glucagon was replaced via the portal vein. An intraportal hyperinsulinemic infusion was initiated at either 2 (INS2) or 5 (INS5) mU^.kg^-1.min^-1. At t=60 min, a 1 mg^.kg^-1.min^-1 portal venous 5-aminoimidazole-4-carboxamide-1--D-ribofuranoside (AICAR) infusion was initiated. Arterial insulin rose ~9-fold and ~27-fold in INS2 and INS5. Glucagon, catecholamines, and cortisol did not change throughout the study. NHGO was completely suppressed prior to t=60 min. Intraportal AICAR stimulated NHGO by 1.9 ± 0.5 and 2.0 ± 0.5 mg^.kg^-1.min^-1in INS2 and INS5, respectively. AICAR stimulated tracer-determined endogenous glucose production similarly in both groups. Intraportal AICAR infusion significantly increased hepatic acetyl-CoA-carboxylase (ACC, Ser⁷⁹) phosphorylation in INS2. Hepatic ACC (Ser⁷⁹) phosphorylation, however, was not increased in INS5. Thus, intraportal AICAR infusion renders hepatic glucose output insensitive to pharmacological insulin. The effectiveness of AICAR in countering the suppressive effect of pharmacological insulin on NHGO occurs even though AICAR-stimulated ACC phosphorylation is completely blocked.