Abstract
The hepatic uptake clearances of 15 ligands with a wide range of permeabilities were determined in rats using two techniques: centrifugal filtration with isolated hepatocytes and the multiple indicator dilution (MID) method with isolated perfused livers. Some of the uptake clearance values were taken from the literature. Uptake clearance values obtained from isolated hepatocytes were extrapolated to that per gram liver (PS inf, cell), assuming that 1 g of liver has 1.3 × 108 cells. The values of PS inf, cell varied from approximately 0.1 to 72 (mL/min/g liver). The values of PS inf, cell were similar to those (PS inf,MID) determined by the MID method for ligands with uptake clearances below approximately 1 mL/min/g liver. However, for the ligands with larger uptake clearances, the PS inf, MID values were lower than the PS inf,cell values and appeared to reach an upper limit (approx. 15–20 mL/min/g liver). The PSinf,cell values of 1-propranolol, tetraphenylphosphonium (TPP+), and diazepam were 72, 43, and 22 mL/min/g liver, respectively, whereas their uptake clearances (PS inf,MID) determined by the MID method were 4 to 10 times lower. One of the possible mechanisms for this discrepancy is that an unstirred water layer, which may exist in Disse's space in isolated perfused livers (and probably under in vivo condition), limits the hepatic uptake rate of ligands with extremely high membrane permeabilities.
Similar content being viewed by others
REFERENCES
M. Rowland, L. Z. Benet, and G. G. Graham. Clearance concepts in pharmacokinetics. J. Pharmacokin. Biopharm. 1:123–136 (1973).
G. R. Wilkinson, and D. G. Shand. A physiological approach to hepatic drug clearance. Clin. Pharmacol. Ther. 18:377–390 (1975).
K. S. Pang, and M. Rowland. Hepatic clearance of drugs I: Theoretical considerations of a “well-stirred” model and a “parallel tube” model. Influence of hepatic blood flow, plasma and blood cell binding, and the hepatocellular enzymatic activity on hepatic drug clearance. J. Pharmacokin. Biopharm. 5:625–653 (1977).
J. R. Gillete. Factors affecting drug metabolism. Ann. N.Y. Acad. Sci. 179:43–66 (1971).
G. Levy and A. Yacobi. Effect of plasma protein binding on elimination of warfarin. J. Pharm. Sci. 63:805–806 (1974).
J. R. Gillete, and K. S. Pang. Theoretic aspects of pharmacokinetic drug interaction. Clin. Pharmacol. Ther. 22:623–639 (1977).
S. Miyauchi, Y. Sugiyama, Y. Sawada, K. Morita, T. Iga, and M. Hanano. Kinetics of hepatic transport of 4-methylumbelliferone in rats: Analysis by multiple indicator dilution method. J. Pharmacokin. Biopharm. 15:25–38 (1987).
L. Schwarz, R. Burr, M. Schwenk, E. Pfaff, and H. Greim. Uptake of taurocholic acid into isolated rat-liver cells. Eur. J. Biochem. 55:617–623 (1975).
C. A. Goresky, W. H. Ziegler, and G. G. Bach. Capillary exchange modeling. Circ. Res. 27:739–764 (1970).
W. M. Pardridge. Unidirectional influx of glutamine and other neutral amino acids into liver of fed and fasted rat in vivo. Am. J. Physiol. 232:E492–E496 (1977).
C. A. Goresky, M. Huet, and J. P. Villeneuve. Blood-exchange and blood flow in the liver. In Hepatology, W. B. Saunders, Philadelphia, 1982, pp. 32–63.
J. Reichen and G. Paumgartner. Uptake of bile acids by perfused rat liver. Am. J. Physiol. 231:734–742 (1976).
Y. Sawada, N. Itoh, Y. Sugiyama, T. Iga, and M. Hanano. Analysis of multiple indicator dilution curves for estimation of renal tubular transport parameters. Comput. Meth. Programs Biomed. 20:51–61 (1985).
S. Miyauchi, Y. Sugiyama, T. Iga, and M. Hanano. Membrane-limited hepatic transport of the conjugative metabolites of 4-methylumbelliferone in rats. J. Pharm Sci. 77:688–692 (1988).
D. Eaton and C. D. Klassen. Carrier-mediated transport of ouabain in isolated hepatocytes. J. Pharmacol. Exp. Ther. 205:480–488 (1978).
P. O. Seglen. Preparation of rat liver cells. III. Enzymatic requirement for tissue dispersion. Exp. Cell Res. 82:391–398 (1973).
S. Miyauchi, Y. Sugiyama, Y. Sawada, T. Iga, and M. Hanano. The influence of glucagon on the hepatic transport of taurocholate in the isolated rat liver perfusion: Kinetic analysis by multiple indicator dilution technique. 108th Japan Pharmaceutical Meeting, Hiroshima, p. 531.
S. Miyauchi, Y. Sugiyama, Y. Sawada, T. Iga, and M. Hanano. The hepatic of 1-propranolol determined by multiple indicator dilution method: The influence of an avid tissue binding of 1-propranolol on its hepatic transport. Xenobio. Metab. Dispos. 3:516–517 (1988).
M. Ichikawa, S. C. Tsao, T. H. Lin, S. Miyauchi, Y. Sawada, T. Iga, M. Hanano, and Y. Sugiyama. Albumin-mediated transport phenomenon” observed for ligands with high membrane permeability: Effect of the unstirred water layer in the Disse's space of rat liver. J. Hepatol. 16:38–49 (1992).
A. G. Ogston and C. F. Phelps. The partition of solute between buffer solutions containing hyaluronic acid. Biochem. J. 78:827–833 (1961).
C. A. Wiederhielm and L. L. Black. Osmotic interaction of plasma proteins with interstitial macromolecules. Am. J. Physiol. 231:638–645 (1976).
E. L. Forker and B. A. Luxon. Effects of unstirred Disse fluid, nonequilibrium binding, and surface-mediated dissociation on hepatic removal of albumin-bound organic anions. Am. J. Physiol. 248:G709–G717 (1985).
L. Bass and S. M. Pond. Hepatic uptake of albumin-bound ligands. In A. Pecile and A. Rescigno (eds.), Pharmacokinetics: Mathematical and Statistical Approaches to Metabolism and Distribution of Chemicals & Drugs, Plenum, London, 1988, pp. 241–265.
P. H. Barry and J. M. Diamond. Effects of unstirred layers on membrane phenomena. Physiol. Rev. 64:763–872 (1984).
R. A. B. Holland, H. Shibata, P. Sheid, and J. Piiper. Rates of uptake and release of oxygen by red blood cells and calculation of the thickness and resistance of an unstirred water layer (abstr.). Proc. Aust. Physiol. Pharmacol. Soc. 13:5 (1982).
S. I. Rubinow. Introduction to Mathematical Biology, Wiley, New York, 1975, p. 209.
A. Blouin, R. O. Bolender, and E. R. Wiebel. Distribution of organelles and membrane between hepatocytes and non-hepatocytes in the rat parenchyma. J. Cell Biol. 72:411–455 (1977).
R. J. Stock, E. V. Cilento and R. S. Mcuskey. A quantitative study of fluorescein isothiocyanate-dextran transport in the microcirculation of the isolated perfused rat liver. Hepatology 9:75–82 (1989).
J. S. Schultz and W. Armstrong. Permeability of interstitial space of muscle (rat diaphragm) to solutes of different molecular weight. J. Pharm. Sci. 67:696–700 (1978).
M. S. Anwer, R. Kroker, and D. Hegner. Cholic acid uptake into isolated hepatocytes. Hoppe-Seyler's Z. Physiol. Chem. 357:1477–1486 (1976).
J. B. Hoek, D. G. Nicholls, and J. R. Williamson. Determination of the mitochondria proton motive force in isolated hepatocytes. J. Biol. Chem. 255:1458–1464 (1980).
S. C. Tsao. Ph.D. thesis, University of Tokyo, Tokyo, 1985.
J. L. Boyer. New concepts of mechanism of hepatocytes bile formation. Physiol. Rev. 60:303–326 (1980).
R. R. Weibel, W. Staubli, H. R. Gnagi, and F. H. Hess. Correlated morphometric and biochemical studies of the liver cell I. Morphometric model, stereologic methods and normal morphometric data for rat liver. J. Cell Biol. 42:68–91 (1969).
M. Wachsein. Enzymatic histochemistry of the liver. Gastroenterology 37:525–537 (1959).
J. J. Gumucio and D. L. Miller. Functional implications of liver cell heterogeneity. Gastroenterology 80:393–403 (1981).
H.-J. Burger, R. Gebhardt, C. Mayer, and D. Mecke. Different capacities for amino acid transport in periportal and perivenous hepatocytes isolated by digitonin/collagenase perfusion. Hepatology 9:22–28 (1989).
L. Bass, S. Keiding, K. Winkler, and N. Tygstrup. Enzymatic elimination of substrates flowing through the intact liver. J. Theor. Biol. 61:393–409 (1976).
A. B. R. Thomson and J. M. Dietschy. Derivation of the equation that describe the effect of unstirred water layers on the kinetic parameters of active transport processes in the intestine. J. Theor. Biol. 64:277–294 (1977).
I. Komiya, J. K. Park, A. Kamani, N. F. Ho, and W. I. Higuchi. Quantitative mechanistic studies in simultaneous fluid flow and intestinal absorption using steroids as model solutes. Int. J. Pharm. 4:249–262 (1980).
Y. Sugiyama, Y. Sawada, T. Iga, and M. Hanano. Reconstruction of in vivo metabolism from in vitro data. In Proceedings of the 2nd International Meeting of the International Society for the Study of Xenobiotics (Kobe, Japan), Taylor & Francis, Philadelphia, 1988, pp. 225–235.
Y. Sugiyama, H. Sato, S. Yanai, D. C. Kim, S. Miyauchi, Y. Sawada, T. Iga, and M. Hanano. Receptor-mediated hepatic clearance of peptide hormones. In D. D. Breimer, D. T. A. Crommelin, and K. K. Midha (eds.), Topics in Pharmaceutical Sciences 1989, Amsterdam Medical Press, Amsterdam, 1989, pp. 429–443.
U. Gartner, R. J. Stockert, W. G. Levine, and A. W. Wolkoff. Effect of nafenopin on the uptake of bilirubin and sulfobro-mophthalein by isolated perfused rat liver. Gastroenterology 83:1163–1169 (1982).
M. S. Anwer, R. Kroker, and D. Hegner. Effect of albumin on bile acid uptake by isolated rat hepatocytes. Is there a common bile acid carrier? Biochem. Biophys. Res. Commun. 73:63–71 (1976).
T. Mizuma, T. Horie, and S. Awazu. The effect of albumin on the uptake of bromosulfophthalein by isolated rat hepatocytes. J. Pharmacobio-Dyn. 8:90–94 (1985).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Miyauchi, S., Sawada, Y., Iga, T. et al. Comparison of the Hepatic Uptake Clearances of Fifteen Drugs with a Wide Range of Membrane Permeabilities in Isolated Rat Hepatocytes and Perfused Rat Livers. Pharm Res 10, 434–440 (1993). https://doi.org/10.1023/A:1018952709120
Issue Date:
DOI: https://doi.org/10.1023/A:1018952709120