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A method was developed to estimate the relative contributions of paracellular and transcellular pathways to the total biliary clearance of sucrose in isolated perfused rat liver. When livers were perfused with a sucrose-containing medium (1 mM), biliary sucrose concentration reached an equilibrium of 165 +/- 27 microM within 10 min, without further significant change up to 40 min. After removal of sucrose from the perfusate, the decrease of the sucrose concentration in bile was found to obey biphasic first-order kinetics, showing a rapid initial decrease (half-life 3.3 +/- 0.5 min) and then a slower decrease (half-life 29.4 +/- 5.7 min). Both phases of decrease were further characterized. Pretreating rats with the cholestatic agents alpha-naphthyl isothiocyanate (ANIT), oestradiol valerate (OV) and colchicine increased the biliary equilibrium concentration and decreased the half-life of the fast phase of the biliary sucrose elimination. The slow phase was unaffected in the livers of ANIT- and OV-treated rats. The slow phase of biliary sucrose efflux was sensitive to colchicine treatment. A close correlation was observed between the slow-phase fraction of the biliary sucrose and the corresponding sucrose content of the liver. By quantitative analysis of the efflux kinetics the relative contribution of the paracellular pathway to the biliary clearance of sucrose was estimated to be 83 +/- 2% in control livers, which increased to about 90% in livers of pretreated animals. These results are important in view of the use of sucrose in evaluating the paracellular-pathway permeability in intra- and extra-hepatic cholestasis.

Quantitative estimation of transcellular and paracellular pathways of biliary sucrose in isolated perfused rat liver