Resveratrol in human hepatoma HepG2 cells: metabolism and inducibility of detoxifying enzymes.
trans-Resveratrol is a polyphenol present in several plant species. Its chemopreventive properties against several diseases have been largely documented. To validate a model for the study of the factors influencing its biological fate at the hepatic level, the metabolism and the efflux of resveratrol were studied in the human hepatoblastoma cell line, HepG2. Comparative high-performance liquid chromatography analysis of cell culture media before and after deconjugation showed that resveratrol was rapidly conjugated; at the concentration of 10 microM, it was entirely metabolized at 8 h of incubation. Two main resveratrol metabolites, monosulfate and disulfate, were identified by atmospheric pressure chemical ionization-mass spectrometry, thanks to their quasi-molecular ion and their characteristic fragmentation. To correlate with the auto-induction of resveratrol metabolism evidenced in HepG2 cells after a pretreatment for 48 h with 10 microM resveratrol, the inducibility of phase II enzymes by resveratrol was studied by real-time quantitative reverse transcriptase-polymerase chain reaction and flow cytometry. Observed, in particular, were an increase in mRNA expression levels of three metabolizing enzymes, two isoforms of UDP-glucuronosyltransferases, UGT1A1 and UGT2B7 (5-fold increased), and a sulfotransferase, ST1E1, in cells pretreated for 24 h with 10 microM resveratrol. These results were correlated with an increase in protein expression, especially after 48 h of treatment. On the other hand, the intracellular resveratrol retention in cells treated with MK571 (3-[[3-[2-(7-chloroquinolin-2-yl)vinyl]phenyl]-(2-dimethylcarbamoylethylsulfanyl)methylsulfanyl] propionic acid), a multidrug resistance-associated protein inhibitor, strongly suggests the involvement of this ABC transporter family in the efflux of resveratrol conjugates from human liver.
Lançon
Allan
A
Institut National de la Santé et de la Recherche Médicale, Unité Mixte de Recherche 866 (Lipides, Nutrition et Cancer, équipe Biochimie Métabolique et Nutritionnelle), Université de Bourgogne, Faculté des Sciences Gabriel, Dijon, France.
Hanet
Nathalie
N
Jannin
Brigitte
B
Delmas
Dominique
D
Heydel
Jean-Marie
JM
Lizard
Gérard
G
Chagnon
Marie-Christine
MC
Artur
Yves
Y
Latruffe
Norbert
N
D000972:Antineoplastic Agents, Phytogenic / Q000378:metabolism / Q000493:pharmacokinetics / Q000494:pharmacology; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D045744:Cell Line, Tumor; D002851:Chromatography, High Pressure Liquid; D004790:Enzyme Induction / Q000187:drug effects; D004798:Enzymes / Q000235:genetics* / Q000378:metabolism; D005434:Flow Cytometry; D014453:Glucuronosyltransferase / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D013058:Mass Spectrometry; D050217:Metabolic Detoxication, Phase II; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D011422:Propionates / Q000494:pharmacology; D011804:Quinolines / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D000077185:Resveratrol; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013267:Stilbenes / Q000378:metabolism* / Q000493:pharmacokinetics* / Q000494:pharmacology; D015238:Sulfotransferases / Q000235:genetics / Q000378:metabolism; D013997:Time Factors; D014316:Tritium; D000099148:UGT1A1 Enzyme
D000972:Antineoplastic Agents, Phytogenic; D004798:Enzymes; D014453:Glucuronosyltransferase; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D011422:Propionates; D011804:Quinolines; D012333:RNA, Messenger; D000077185:Resveratrol; D013267:Stilbenes; D015238:Sulfotransferases; D014316:Tritium; D000099148:UGT1A1 Enzyme; C059141:verlukast; C491474:UGT2B7 protein, human; C020122:estrone sulfotransferase