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Metabolism and Disposition of Resveratrol in the Isolated Perfused Rat Liver: Role of Mrp2 in the Biliary Excretion of Glucuronides

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ABSTRACT

In this study, the hepatic metabolism and transport system for resveratrol was examined in isolated perfused livers from Wistar and Mrp2‐deficient TR rats. Based on extensive metabolism to six glucuronides and sulfates (M1–M6), the hepatic extraction ratio and clearance of resveratrol was very high in Wistar and TR rats (E: 0.998 vs. 0.999; Cl: 34.9 mL/min vs. 36.0 mL/min). However, biliary excretion and efflux of conjugates differs greatly in TR rats. While cumulative biliary excretion of the glucuronides M1, M2, M3, and M5 dropped dramatically to 0–6%, their efflux into perfusate increased by 3.6‐, 1.8‐, 2.5‐, and 1.5‐fold. In contrast, biliary secretion of the sulfates M4 and M6 was partially maintained in the Mrp2‐deficient rats (61% and 39%) with a concomitant decline of their efflux into perfusate by 33.2% and 78.1%. This indicates that Mrp2 exclusively mediates the biliary excretion of resveratrol glucuronides but only partly that of sulfates. Cumulative secretion of unconjugated resveratrol into bile of TR rats was only reduced by 40%, and into perfusate by 19%, suggesting only a minor role of Mrp2 in resveratrol elimination. In summary, resveratrol was dose‐dependently metabolized to several conjugates whereby the canalicular transporter Mrp2 selectively mediated the biliary excretion of glucuronides. © 2007 Wiley‐Liss, Inc. and the American Pharmacists Association J Pharm Sci 97:1615–1628, 2008

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INTRODUCTION

Resveratrol (trans‐3,5,4′‐trihydroxystilbene) is a polyphenolic phytoalexin found in grapes, fruits, and root extracts of Polygonum cuspidatum, an important constituent of Chinese folk medicine used to fight liver, skin, and circulatory diseases.1, 2 A great number of studies carried out in cellular and animal models have already demonstrated that resveratrol regulates many biological functions as it modulates lipid and lipoprotein metabolism,3, 4 inhibits platelet aggregation,5., 6., 7. alters

Materials

trans‐Resveratrol, sulfatase type V from Limpets (Patella vulgata), and β‐glucuronidase type B‐3 from bovine liver were obtained from Sigma (Munich, Germany). 14C‐Taurocholic acid was from NEN Life Science Products (Perkin‐Elmer, Wellesley, MA), bromosulfophthalein from Merck (Darmstadt, Germany). Methanol and water were of HPLC grade (Merck). All other chemicals and solvents were of analytical grade and used without further purification.

Liver Perfusion

The study was approved by the Animal Experimentation

Metabolism and Disposition of Resveratrol in Wistar Rats

Application of resveratrol (5–40 µM) to the liver of Wistar rats moderately stimulated bile flow between 6% and 30%. Choleresis reached its peak values at 10–15 min after addition of resveratrol to the perfusion medium (mean maximum values: 0.84 ± 0.09; 0.91 ± 0.11; 1.1 ± 0.07; 1.2 ± 0.13 µg/g liver/min, respectively), indicating moderate solvent drag by the concentrated excretion of resveratrol and its metabolites into bile. Afterwards, bile flow slowly declined reaching initial values after 90 

DISCUSSION

In the present study, hepatic biotransformation and hepatobiliary transport of resveratrol and its metabolites was investigated by perfusing isolated rat livers of male Wistar rats with increasing concentrations of resveratrol (5–40 µM) which were chosen based on daily intakes of resveratrol as beverage (red wine) or as dietary supplement (5–25 mg/day). During 90 min the perfused liver extracted more than 99% of applied resveratrol with steady‐state concentrations attained within 5 min after

Acknowledgements

This study was supported by a grant from the Jubiläumsfonds der Österreichischen Nationalbank 9894 (W.J.) and by the Fonds zur Förderung der Wissenschaftlichen Forschung des Bürgermeisters der Bundeshauptstadt Wien (2296; T.S.).

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