Characterization of phloretin-sensitive urea export from the perfused rat liver

In single pass perfused rat liver, rapid osmotic water shifts across the plasma membrane in response to hyperosmolar urea were followed by monitoring liver mass and transient concentrating or diluting effects on Na+ concentration in effluent perfusate. Sudden addition or removal of hyperosmolar urea (200mM, resulting in a step change of the perfusate osmolarity from 305 to 505 mosmol/l) had little effect on liver mass or Na+ activity in the effluent perfusate, suggesting that urea equilibrated at a rate similar to that of water across the liver plasma membrane. When, however, phloretin (0.2mM) was present, sudden addition (removal) of urea (200mM) induced within seconds a marked and transient decrease (increase) of both liver mass and effluent Na+ concentration, suggestive of transient osmotic water shifts out of/into the cells. Although to a lesser extent, comparable effects were induced when urea was added/removed in the presence of the phloretin-related phenol compounds 2,4,6-trihydroxyacetophenone (5mM) and 2,4,5-trihydroxybutyrophenone (5mM). Phloretin-induced inhibition of urea export from livers preloaded with [14C]urea was reversible, and no saturation of urea transport was found at concentrations up to 200mM. In contrast to [14C]urea transport, [3H]water transport across the plasma membrane was not affected by phloretin. The data indicate that urea export across the hepatocyte plasma membrane is almost as fast as water export. The urea transport mechanism is sensitive to phloretin and other phenol compounds, works with high capacity and is distinct from the water-transporting system. The regulation of this putative transport mechanism and its relevance for hepatic nitrogen metabolism remain to be established.