Effects of bile duct ligation and dietary change on hepatic carbohydrate metabolism in rats were studied by analyzing the levels of fructose 2,6-bisphosphate (fructose-2,6-P2) and other metabolites. There was no significant difference in the levels of fructose-2,6-P2 and glycolytic intermediates between rats with and without bile duct ligation when rats were in a fed state. However, the levels of fructose-2,6-P2, hexose monophosphates, and triose phosphates increased and the level of plasma glucose decreased in rats with bile duct ligation in a starved state. The crossover analysis of gluconeogenic intermediates in rats with bile duct ligation showed accumulations of hexose monophosphates together with fructose-2,6-P2 in spite of the starved state of rats, indicating an impaired conversion of glucose 6-phosphate to glucose. Despite the activation of the fructose 6-phosphate-fructose 1,6-bisphosphate futile cycle as indicated by these findings, the energy charge levels were not reduced. The block at the level of glucose production may explain the relative predominance of insulin effect on glycolysis.