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Interplay between MRP Inhibition and Metabolism of MRP Inhibitors: The Case of Curcumin

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Author: Wortelboer, H.M. · Usta, M. · Velde, A.E. van der · Boersma, M.G. · Spenkelink, B. · Zanden, J.J. van · Rietjens, I.M.C.M. · Bladeren, P.J. van · Cnubben, N.H.P.
Type:article
Date:2003
Source:Chemical Research in Toxicology, 12, 16, 1642-1651
Identifier: 237512
doi: doi:10.1021/tx034101x
Keywords: Biology · Physiological Sciences · curcumin · glutathione · glutathionylcurcumin · multidrug resistance protein 1 · multidrug resistance protein 2 · protein inhibitor · unclassified drug · animal cell · article · controlled study · drug effect · IC 50 · inhibition kinetics · insect cell · intracellular transport · membrane vesicle · metabolism · monolayer culture · nonhuman · protein expression · protein transport · Animals · ATP-Binding Cassette Transporters · Baculoviridae · Cell Line · Curcumin · Cyclosporine · Dinitrochlorobenzene · Dose-Response Relationship, Drug · Drug Resistance, Multiple · Ethacrynic Acid · Fluoresceins · Glutathione · Glutathione Transferase · Humans · P-Glycoprotein · P-Glycoproteins · Propionates · Quinolines · Recombinant Proteins · Spodoptera · Animalia · Apus apus · Insecta

Abstract

The multidrug resistance proteins MRP1 and MRP2 are efflux transporters with broad substrate specificity, including glutathione, glucuronide, and sulfate conjugates. In the present study, the interaction of the dietary polyphenol curcumin with MRP1 and MRP2 and the interplay between curcumin-dependent MRP inhibition and its glutathione-dependent metabolism were investigated using two transport model systems. In isolated membrane vesicles of MRP1- and MRP2-expressing Sf9 cells, curcumin clearly inhibited both MRP1- and MRP2-mediated transport with IC50 values of 15 and 5 μM, respectively. In intact monolayers of MRP1 overexpressing Madin-Darby canine kidney (MDCKII-MRP1) cells, curcumin also inhibited MRPl-mediated activity, although with a 3-fold higher IC50 value than the one observed in the vesicle model. Interestingly, MRP2-mediated activity was hardly inhibited in intact monolayers of MRP2-overexpressing MDCKII (MDCKII-MRP2) cells upon exposure to curcumin, whereas the IC50 value in the vesicle incubations was 5 μM. The difference in extent of inhibition of the MRPs by curcumin in isolated vesicles as compared to intact cells, observed especially for MRP2, was shown to be due to a swift metabolism of curcumin to two glutathione conjugates in the MDCKII cells. Formation of both glutathione conjugates was about six times higher in the MDCKII-MRP2 cells as compared with the MDCKII-MRP1 cells, a phenomenon that could be ascribed to the significantly lower glutathione levels in the cell line. The efflux of both conjugates, identified in the present study as monoglutathionyl curcumin conjugates, was demonstrated to be mediated by both MRP1 and MRP2. From dose-response curves with Sf9 membrane vesicles, glutathionylcurcumin conjugates appeared to be less potent inhibitors of MRP1 and MRP2 than their parent compound curcumin. In conclusion, curcumin clearly inhibits both MRP1- and MRP2-mediated transport, but the glutathione-dependent metabolism of curcumin plays a crucial role in the ultimate level of inhibition of MRP-mediated transport that can be achieved in a cellular system. This complex interplay between MRP inhibition and metabolism of MRP inhibitors, the latter affecting the ultimate potential of a compound for cellular MRP inhibition, may exist not only for a compound like curcumin but also for many other MRP inhibitors presently or previously developed on the basis of vesicle studies.