Regulation of biotransformation systems and ABC transporters by benznidazole in HepG2 cells

• Verfasst von: Rigalli, Juan Pablo [VerfasserIn]
• Verfasst von: Theile, Dirk [VerfasserIn]
• Verfasst von: Weiß, Johanna [VerfasserIn]
• Titel: Regulation of biotransformation systems and ABC transporters by benznidazole in HepG2 cells
• Titelzusatz: involvement of pregnane X-receptor
• Verf.angabe: Juan P. Rigalli, Virginia G. Perdomo, Marcelo G. Luquita, Silvina S.M. Villanueva, Agostina Arias, Dirk Theile, Johanna Weiss, Aldo D. Mottino, María L. Ruiz, Viviana A. Catania
• Jahr: 2012
• Fussnoten: Published December 13, 2012 ; Gesehen am 26.06.2018
• Titel Quelle: Enthalten in: Public Library of SciencePLoS neglected tropical diseases
• Ort Quelle: Lawrence, Kan. : PLoS, 2007
• Jahr Quelle: 2012
• Band/Heft Quelle: 6(2012,12) Artikel-Nummer e1951, 10 Seiten
• ISSN Quelle: 1935-2735
• DOI: doi:10.1371/journal.pntd.0001951
• Datenträger: Online-Ressource
• Sprache: eng
• Sach-SW: Cell membranes
• Sach-SW: Drug metabolism
• Sach-SW: High performance liquid chromatography
• Sach-SW: Messenger RNA
• Sach-SW: Protein expression
• Sach-SW: Small interfering RNAs
• Sach-SW: Trypanosoma cruzi
• Sach-SW: Xenobiotic metabolism
• K10plus-PPN: 1576840271

Abstract

Background Benznidazole (BZL) is the only antichagasic drug available in most endemic countries. Its effect on the expression and activity of drug-metabolizing and transporter proteins has not been studied yet. Methodology/Principal Findings Expression and activity of P-glycoprotein (P-gp), Multidrug resistance-associated protein 2 (MRP2), Cytochrome P450 3A4 (CYP3A4), and Glutathione S-transferase (GST) were evaluated in HepG2 cells after treatment with BZL. Expression was estimated by immunoblotting and real time PCR. P-gp and MRP2 activities were estimated using model substrates rhodamine 123 and dinitrophenyl-S-glutathione (DNP-SG), respectively. CYP3A4 and GST activities were evaluated through their abilities to convert proluciferin into luciferin and 1-chloro-2,4-dinitrobenzene into DNP-SG, respectively. BZL (200 µM) increased the expression (protein and mRNA) of P-gp, MRP2, CYP3A4, and GSTπ class. A concomitant enhancement of activity was observed for all these proteins, except for CYP3A4, which exhibited a decreased activity. To elucidate if pregnane X receptor (PXR) mediates BZL response, its expression was knocked down with a specific siRNA. In this condition, the effect of BZL on P-gp, MRP2, CYP3A4, and GSTπ protein up-regulation was completely abolished. Consistent with this, BZL was able to activate PXR, as detected by reporter gene assay. Additional studies, using transporter inhibitors and P-gp-knock down cells, demonstrated that P-gp is involved in BZL extrusion. Pre-treatment of HepG2 cells with BZL increased its own efflux, as a consequence of P-gp up-regulation. Conclusions/Significance Modifications in the activity of biotransformation and transport systems by BZL may alter the pharmacokinetics and efficiency of drugs that are substrates of these systems, including BZL itself.