We examined the role of the hepatic vagus nerve in hepatic and peripheral glucose metabolism. To assess endogenous glucose production (EGP), hepatic uptake of first-pass glucose infused intraportally (HGU), and the metabolic clearance rate of glucose (MCR), rats were subjected to hepatic vagotomy (HV, n = 7) or sham operation (SH, n = 8), after 10 days, they were then subjected to a euglycemic-hyperinsulinemic clamp together with a portal glucose load in the 24-hour fasting state. Metabolic parameters were determined by the dual-tracer method using stable isotopes. During the experiment, [6,6-²H₂]glucose was continuously infused into the peripheral vein. to maintain euglycemia (4.5 mmol/L), insulin (54 pmol · kg⁻¹ · min⁻¹) and glucose were infused peripherally after the 90-minute tracer equilibration and 30-minute basal periods, and glucose containing 5% enriched [U-¹³C]glucose was infused intraportally (50 μmol · kg⁻¹ · min⁻¹) for 120 minutes (clamp period). EGP was significantly higher in HV rats versus SH rats during the basal period (64.3 ± 7.6 v 43.6 ± 5.3 μmol · kg⁻¹ · min⁻¹, P < .005) and was comparable to EGP in SH rats during the clamp period (9.3 ± 21.5 v 1.1 ± 11.7 μmol · kg⁻¹ · min⁻¹). HGU was reduced in HV rats compared with SH rats during portal glucose infusion (5.9 ± 2.4 v 10.1 ± 3.2 μmol · kg⁻¹ · min⁻¹). The MCR in HV rat was significantly higher than in SH rats in the basal period (11.0 ± 2.0 v 7.9 ± 0.8 mL · kg⁻¹ · min⁻¹, P < 0.01 and was comparable to the MCR in SH rats during the clamp eriod (41.9 ± 10.0 and 36.6 ± 5.7 mL · kg⁻¹ · min⁻¹. We conclude that innervation of the hepatic vagus nerve is important for the regulation of hepatic glucose production in the postabsorptive state and HGU in the postprandial state.