Skip to main content

Advertisement

SpringerLink
Log in

Allyl isothiocyanate (AITC) inhibits pregnane X receptor (PXR) and constitutive androstane receptor (CAR) activation and protects against acetaminophen- and amiodarone-induced cytotoxicity

  • Molecular Toxicology
  • Published:
Archives of Toxicology Aims and scope Submit manuscript

Abstract

Antagonizing the action of the pregnane X receptor (PXR) may have important clinical implications for preventing inducer–drug interactions and improving therapeutic efficacy. We identified a widely distributed isothiocyanate, allyl isothiocyanate (AITC), which acts as an effective antagonist of the nuclear receptor pregnane X receptor (PXR, NR1I2) and constitutive androstane receptor (CAR, NR1I3). HepG2 cells were used to assay reporter function, mRNA levels, and protein expression. Catalytic activities of the PXR and CAR target genes, CYP3A4 and CYP2B6, respectively, were also assessed in differentiated HepaRG cells. Protective effects of AITC on rifampin-induced cytotoxicity were observed, and transient transfection assays showed that AITC was able to effectively attenuate the agonist effects of rifampin and CITCO on human PXR and CAR activity, respectively. AITC-mediated reduction in the transcriptional activity of PXR and CAR correlated well with the suppression of CYP3A4 and CYP2B6 expression in HepG2 cells, which reflected the reduced catalytic activities of both of these genes following AITC treatment in differentiated HepaRG cells. Furthermore, AITC disrupts the co-regulations of PXR with several important co-regulators. Furthermore, the antagonist effect of AITC against PXR was found in HepaRG cells upon addition of acetaminophen (APAP) and amiodarone, indicating that AITC protects cells from drug-induced cytotoxicity. Taken together, our results show that AITC inhibits the transactivation effects of PXR and CAR and reduces the expression and function of CYP3A4 and CYP2B6. Additionally, AITC reversed the cytotoxic effects of APAP and amiodarone induced by PXR ligand. Results from this study suggest that AITC could be a powerful agent for reducing potentially dangerous interactions between transcriptional inducers of CYP enzymes and therapeutic drugs.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

Abbreviations

AITC:

Allyl isothiocyanate

CAR:

Constitutive androstane receptor

CITCO:

6-(4-Chlorophenyl)imidazo[2,1-b][1,3]thiazole-5-carbaldehydeO-(3,4-dichlorobenzyl)-oxime

CYP450:

Cytochrome P450

DMEs:

Drug-metabolizing enzymes

DMSO:

Dimethylsulfoxide

DR:

Direct repeat

ER:

Everted repeat

GR:

Glucocorticoid receptor

HNF4α:

Hepatocyte nuclear factor 4α

PCN:

Pregnenolone 16α-carbonitrile

PGC-1α:

Peroxisome proliferator-activated receptor-gamma coactivator 1α

PXR:

Pregnane X receptor

RXRα:

Retinoic X receptor α

SRC-1:

Steroid receptor coactivator-1

TCPOBOP:

1,4-Bis[2-(3,5-dichloropyridyloxy)]benzene

References

  • Ahlin G, Hilgendorf C, Karlsson J, Szigyarto CA, Uhlén M, Artursson P (2009) Endogenous gene and protein expression of drug-transporting proteins in cell lines routinely used in drug discovery programs. Drug Metab Dispos 37:2275–2283

    Article  CAS  PubMed  Google Scholar 

  • Bauer B, Yang X, Hartz AM, Olson ER, Zhao R, Kalvass JC, Pollack GM, Miller DS (2006) In vivo activation of human pregnane X receptor tightens the blood-brain barrier to methadone through P-glycoprotein up-regulation. Mol Pharmacol 70:1212–1219

    Article  CAS  PubMed  Google Scholar 

  • Beckett GJ, Chapman BJ, Dyson EH, Hayes JD (1985) Plasma glutathione S-transferase measurements after paracetamol overdose: evidence for early hepatocellular damage. Gut 26:26–31

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Bertilsson G, Heidrich J, Svensson K, Asman M, Jendeberg L, Sydow-Backman M, Ohlsson R, Postlind H, Blomquist P, Berkenstam A (1998) Identification of a human nuclear receptor defines a new signaling pathway for CYP3A induction. Proc Natl Acad Sci USA 95:12208–12213

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Boelsterli UA, Lim PL (2007) Mitochondrial abnormalities—a link to idiosyncratic drug hepatotoxicity? Toxicol Appl Pharmacol 220:92–107

    Article  CAS  PubMed  Google Scholar 

  • Bollard M, Stribbling S, Mitchell S, Caldwell J (1997) The disposition of allyl isothiocyanate in the rat and mouse. Food Chem Toxicol 35:933–943

    Article  CAS  PubMed  Google Scholar 

  • Chen T (2010) Overcoming drug resistance by regulating nuclear receptors. Adv Drug Deliv Rev 62:1257–1264

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Chen T, Tompkins LM, Li L, Li H, Kim G, Zheng Y, Wang H (2010a) A single amino acid controls the functional switch of human constitutive androstane receptor (CAR) 1 to the xenobiotic-sensitive splicing variant CAR3. J Pharmacol Exp Ther 332:106–115

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Chen Y, Tang Y, Robbins GT, Nie D (2010b) Camptothecin attenuates cytochrome P450 3A4 induction by blocking the activation of human pregnane X receptor. J Pharmacol Exp Ther 334:999–1008

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Chen Y, Tang Y, Guo C, Wang J, Boral D, Nie D (2012) Nuclear receptors in the multidrug resistance through the regulation of drug-metabolizing enzymes and drug transporters. Biochem Pharmacol 83:1112–1126

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Cheng J, Ma X, Krausz KW, Idle JR, Gonzalez FJ (2009) Rifampicin-activated human pregnane X receptor and CYP3A4 induction enhance acetaminophen-induced toxicity. Drug Metab Dispos 37:1611–1621

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Cheng J, Shah YM, Gonzalez FJ (2012) Pregnane X receptor as a target for treatment of inflammatory bowel disorders. Trends Pharmacol Sci 33:323–330

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Conney AH (2003) Induction of drug-metabolizing enzymes: a path to the discovery of multiple cytochromes P450. Ann Rev Pharmacol Toxicol 43:1–30

    Article  CAS  Google Scholar 

  • Dahlin DC, Miwa GT, Lu AY, Nelson SD (1984) N-acetyl-p-benzoquinone imine: a cytochrome P-450-mediated oxidation product of acetaminophen. Proc Natl Acad Sci USA 81:1327–1331

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • EFSA (2010) Scientific Opinion on the safety of allyl isothiocyanate for the proposed uses as a food additive. EFSA J 8:1943

    Google Scholar 

  • Flanagan RJ, Storey GC, Holt DW, Farmer PB (1982) Identification and measurement of desethylamiodarone in blood plasma specimens from amiodarone-treated patients. J Pharm Pharmacol 34:638–643

    Article  CAS  PubMed  Google Scholar 

  • Gao J, Xie W (2012) Targeting xenobiotic receptors PXR and CAR for metabolic diseases. Trends Pharmacol Sci 33:552–558

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Goodwin B, Hodgson E, Liddle C (1999) The orphan human pregnane X receptor mediates the transcriptional activation of CYP3A4 by rifampicin through a distal enhancer module. Mol Pharmacol 56:1329–1339

    CAS  PubMed  Google Scholar 

  • Goodwin B, Moore LB, Stoltz CM, McKee DD, Kliewer SA (2001) Regulation of the human CYP2B6 gene by the nuclear pregnane X receptor. Mol Pharmacol 60:427–431

    CAS  PubMed  Google Scholar 

  • Guengerich FP (2008) Cytochrome P450 and chemical toxicology. Chem Res Toxicol 21:70–83

    Article  CAS  PubMed  Google Scholar 

  • Guengerich FP, MacDonald JS (2007) Applying mechanisms of chemical toxicity to predict drug safety. Chem Res Toxicol 20:344–369

    Article  CAS  PubMed  Google Scholar 

  • Guillouzo A, Corlu A, Aninat C, Glaise D, Morel F, Guguen-Guillouzo C (2007) The human hepatoma HepaRG cells: a highly differentiated model for studies of liver metabolism and toxicity of xenobiotics. Chem Biol Interact 168:66–73

    Article  CAS  PubMed  Google Scholar 

  • Guo GL, Moffit JS, Nicol CJ, Ward JM, Aleksunes LA, Slitt AL, Kliewer SA, Manautou JE, Gonzalez FJ (2004) Enhanced acetaminophen toxicity by activation of the pregnane X receptor. Toxicol Sci 82:374–380

    Article  CAS  PubMed  Google Scholar 

  • Ha HR, Bigler L, Wendt B, Maggiorini M, Follath F (2005) Identification and quantitation of novel metabolites of amiodarone in plasma of treated patients. Eur J Pharm Sci 24:271–279

    Article  CAS  PubMed  Google Scholar 

  • Hancock JF (1992) COS Cell Expression. Methods Mol Biol 8:153–158

    CAS  PubMed  Google Scholar 

  • Healan-Greenberg C, Waring JF, Kempf DJ, Blomme EA, Tirona RG, Kim RB (2008) A human immunodeficiency virus protease inhibitor is a novel functional inhibitor of human pregnane X receptor. Drug Metab Dispos 36:500–507

    Article  CAS  PubMed  Google Scholar 

  • Hosomi H, Akai S, Minami K, Yoshikawa Y, Fukami T, Nakajima M, Yokoi T (2010) An in vitro drug-induced hepatotoxicity screening system using CYP3A4-expressing and gamma-glutamylcysteine synthetase knockdown cells. Toxicol In Vitro 24:1032–1038

    Article  CAS  PubMed  Google Scholar 

  • Hu ZP, Yang XX, Chen X, Cao J, Chan E, Duan W, Huang M, Yu XQ, Wen JY, Zhou SF (2007) A mechanistic study on altered pharmacokinetics of irinotecan by St John’s wort. Curr Drug Metab 8:157–171

    Article  CAS  PubMed  Google Scholar 

  • Itoh M, Nakajima M, Higashi E, Yoshida R, Nagata K, Yamazoe Y, Yokoi T (2006) Induction of human CYP2A6 is mediated by the pregnane X receptor with peroxisome proliferator-activated receptor-gamma coactivator 1 alpha. J Pharmacol Exp Ther 319:693–702

    Article  CAS  PubMed  Google Scholar 

  • Jacobs MN, Nolan GT, Hood SR (2005) Lignans, bacteriocides and organochlorine compounds activate the human pregnane X receptor (PXR). Toxicol Appl Pharmacol 209:123–133

    Article  CAS  PubMed  Google Scholar 

  • James LP, Mayeux PR, Hinson JA (2003) Acetaminophen-induced hepatotoxicity. Drug Metab Dispos 31:1499–1506

    Article  CAS  PubMed  Google Scholar 

  • Kawamoto T, Sueyoshi T, Zelko I, Moore R, Washburn K, Negishi M (1999) Phenobarbital-responsive nuclear translocation of the receptor CAR in induction of the CYP2B gene. Mol Cell Biol 19:6318–6322

    CAS  PubMed Central  PubMed  Google Scholar 

  • Kliewer SA, Goodwin B, Willson TM (2002) The nuclear pregnane X receptor: a key regulator of xenobiotic metabolism. Endocr Rev 23:687–702

    Article  CAS  PubMed  Google Scholar 

  • Kostrubsky VE, Szakacs JG, Jeffery EH, Woods SG, Bement WJ, Wrighton SA, Sinclair PR, Sinclair JF (1997) Protection of ethanol-mediated acetaminophen hepatotoxicity by triacetyloleandomycin, a specific inhibitor of CYP3A. Ann Clin Lab Sci 27:57–62

    CAS  PubMed  Google Scholar 

  • Krausova L, Stejskalova L, Wang H, Vrzal R, Dvorak Z, Mani S, Pavek P (2011) Metformin suppresses pregnane X receptor (PXR)-regulated transactivation of CYP3A4 gene. Biochem Pharmacol 82:1180–1771

    Article  Google Scholar 

  • Krul C, Humblot C, Philippe C, Vermeulen M, van Nuenen M, Havenaar R, Rabot S (2002) Metabolism of sinigrin (2-propenyl glucosinolate) by the human colonic microflora in a dynamic in vitro large-intestinal model. Carcinogenesis 23:1009–1016

    Article  CAS  PubMed  Google Scholar 

  • Lake BG, Renwick AB, Cunninghame ME, Price RJ, Surry D, Evans DC (1998) Comparison of the effects of some CYP3A and other enzyme inducers on replicative DNA synthesis and cytochrome P450 isoforms in rat liver. Toxicology 131:9–20

    Article  CAS  PubMed  Google Scholar 

  • Lehmann JM, McKee DD, Watson MA, Willson TM, Moore JT, Kliewer SA (1998) The human orphan nuclear receptor PXR is activated by compounds that regulate CYP3A4 gene expression and cause drug interactions. J Clin Invest 102:1016–1023

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Lim YP, Ma CY, Liu CL, Lin YH, Hu ML, Chen JJ, Hung DZ, Hsieh WT, Huang JD (2012) Sesamin: a naturally occurring lignan inhibits CYP3A4 by antagonizing the pregnane X receptor activation. Evid Based Complement Alternat Med 2012:242810

    PubMed Central  PubMed  Google Scholar 

  • Liu CL, Lim YP, Hu ML (2012) Fucoxanthin attenuates rifampin-induced cytochrome P450 3A4 (CYP3A4) and multiple drug resistance 1 (MDR1) gene expression through pregnane X receptor (PXR)-mediated pathways in human hepatoma HepG2 and colon adenocarcinoma LS174T cells. Mar Drugs 10:242–257

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Lo WS, Lim YP, Chen CC, Hsu CC, Souček P, Yun CH, Xie W, Ueng YF (2012) A dual function of the furanocoumarin chalepensin in inhibiting Cyp2a and inducing Cyp2b in mice: the protein stabilization and receptor-mediated activation. Arch Toxicol 86:1927–1938

    Article  CAS  PubMed  Google Scholar 

  • Mahamuni SP, Khose RD, Menaa F, Badole SL (2012) Therapeutic approaches to drug targets in hyperlipidemia. BioMedicine 2:137–146

    Article  Google Scholar 

  • Mason JW (1987) Amiodarone. N Engl J Med 316:455–466

    Article  CAS  PubMed  Google Scholar 

  • Matsuda H, Ochi M, Nagatomo A, Yoshikawa M (2007) Effects of allyl isothiocyanate from horseradish on several experimental gastric lesions in rats. Eur J Pharmacol 561:172–181

    Article  CAS  PubMed  Google Scholar 

  • McCarthy TC, Pollak PT, Hanniman EA, Sinal CJ (2004) Disruption of hepatic lipid homeostasis in mice after amiodarone treatment is associated with peroxisome proliferator activated receptor alpha target gene activation. J Pharmacol Exp Ther 311:864–873

    Article  CAS  PubMed  Google Scholar 

  • Moore LB, Maglich JM, McKee DD, Wisely B, Willson TM, Kliewer SA, Lambert MH, Moore JT (2002) Pregnane X receptor (PXR), constitutive androstane receptor (CAR), and benzoate X receptor (BXR) define three pharmacologically distinct classes of nuclear receptors. Mol Endocrinol 16:977–986

    Article  CAS  PubMed  Google Scholar 

  • Moya M, Gomez-Lechon MJ, Castell JV, Jover R (2010) Enhanced steatosis by nuclear receptor ligands: a study in cultured human hepatocytes and hepatoma cells with a characterized nuclear receptor expression profile. Chem Biol Interact 184:376–387

    Article  CAS  PubMed  Google Scholar 

  • Munday R, Munday CM (2002) Selective induction of phase II enzymes in the urinary bladder of rats by allyl isothiocyanate, a compound derived from Brassica vegetables. Nutr Cancer 44:52–59

    Article  CAS  PubMed  Google Scholar 

  • Nielsen PV, Rios R (2000) Inhibition of fungal growth on bread by volatile components from spices and herbs, and the possible application in active packaging, with special emphasis on mustard essential oil. Int J Food Microbiol 60:219–229

    Article  CAS  PubMed  Google Scholar 

  • Pascussi JM, Robert A, Nguyen M, Walrant-Debray O, Garabedian M, Martin P, Pineau T, Saric J, Navarro F, Maurel P, Vilarem MJ (2005) Possible involvement of pregnane X receptor-enhanced CYP24 expression in drug-induced osteomalacia. J Clin Invest 115:177–186

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Potter WZ, Thorgeirsson SS, Jollow DJ, Mitchell JR (1974) Acetaminophen induced hepatic necrosis. V. Correlation of hepatic necrosis, covalent binding and glutathione depletion in hamsters. Pharmacology 12:129–143

    Article  CAS  PubMed  Google Scholar 

  • Reimers D, Jezek A (1971) The simultaneous use of rifampicin and other antitubercular agents with oral contraceptives. Prax Pneumol 25:255–262

    CAS  PubMed  Google Scholar 

  • Rigalli JP, Ruiz ML, Perdomo VG, Villanueva SS, Mottino AD, Catania VA (2011) Pregnane X receptor mediates the induction of P-glycoprotein by spironolactone in HepG2 cells. Toxicology 285:18–24

    Article  CAS  PubMed  Google Scholar 

  • Rodriguez-Antona C, Donato MT, Boobis A, Edwards RJ, Watts PS, Castell JV, Gomez-Lechon MJ (2002) Cytochrome P450 expression in human hepatocytes and hepatoma cell lines: molecular mechanisms that determine lower expression in cultured cells. Xenobiotica 32:505–520

    Article  CAS  PubMed  Google Scholar 

  • Schulman IG (2010) Nuclear receptors as drug targets for metabolic disease. Adv Drug Deliv Rev 62:1307–1315

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Shin IS, Masuda H, Naohide K (2004) Bactericidal activity of wasabi (Wasabia japonica) against Helicobacter pylori. Int J Food Microbiol 94:255–261

    Article  PubMed  Google Scholar 

  • Staudinger JL, Goodwin B, Jones SA, Hawkins-Brown D, MacKenzie KI, LaTour A, Liu Y, Klaassen CD, Brown KK, Reinhard J, Willson TM, Koller BH, Kliewer SA (2001) The nuclear receptor PXR is a lithocholic acid sensor that protects against liver toxicity. Proc Natl Acad Sci USA 98:3369–3374

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Sueyoshi T, Kawamoto T, Zelko I, Honkakoski P, Negishi M (1999) The repressed nuclear receptor CAR responds to phenobarbital in activating the human CYP2B6 gene. J Biol Chem 274:6043–6046

    Article  CAS  PubMed  Google Scholar 

  • Sultana T, Porter NG, Savage GP, McNeil DL (2003) Comparison of isothiocyanate yield from wasabi rhizome tissues grown in soil or water. J Agric Food Chem 51:3586–3591

    Article  CAS  PubMed  Google Scholar 

  • Synold TW, Dussault I, Forman BM (2001) The orphan nuclear receptor SXR coordinately regulates drug metabolism and efflux. Nat Med 7:584–590

    Article  CAS  PubMed  Google Scholar 

  • Takezawa T, Matsunaga T, Aikawa K, Nakamura K, Ohmori S (2012) Lower expression of HNF4α and PGC1α might impair rifampicin-mediated CYP3A4 induction under conditions where PXR is overexpressed in human fetal liver cells. Drug Metab Pharmacokinet 27:430–438

    Article  CAS  PubMed  Google Scholar 

  • TNO (Netherlands Organisation for Applied Scientific Research) (2009) VCF volatile compounds in food: database/Nijssen, L.M.; Ingen-Visscher, C.A. van; Donders, J.J.H. [eds]. – Version 12.1 –Zeist (The Netherlands): TNO Quality of Life, 1963–2009. http://www.vcf-online.nl/VcfHome.cfm

  • Wagner AE, Boesch-Saadatmandi C, Dose J, Schultheiss G, Rimbach G (2012) Anti-inflammatory potential of allyl isothiocyanate − role of Nrf2, NF-κB and microRNA-155. J Cell Mol Med 16:836–843

    Article  CAS  PubMed  Google Scholar 

  • Walko CM, Combest AJ, Spasojevic I, Yu AY, Bhushan S, Hull JH, Hoskins J, Armstrong D, Carey L, Collicio F, Dees EC (2012) The effect of a prepitant and race on the pharmacokinetics of cyclophosphamide in breast cancer patients. Cancer Chemother Pharmacol 69:1189–1196

    Article  CAS  PubMed  Google Scholar 

  • Wang H, Tompkins LM (2008) CYP2B6: new insights into a historically overlooked cytochrome P450 isozyme. Curr Drug Metab 9:598–610

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Wang H, Faucette SR, Gilbert D, Jolley SL, Sueyoshi T, Negishi M, LeCluyse EL (2003) Glucocorticoid receptor enhancement of pregnane X receptor-mediated CYP2B6 regulation in primary human hepatocytes. Drug Metab Dispos 31:620–630

    Article  CAS  PubMed  Google Scholar 

  • Wang H, Huang H, Li H, Teotico DG, Sinz M, Baker SD, Staudinger J, Kalpana G, Redinbo MR, Mani S (2007) Activated pregnenolone X-receptor is a target for ketoconazole and its analogs. Clin Cancer Res 13:2488–2495

    Article  CAS  PubMed  Google Scholar 

  • Wang H, Li H, Moore LB, Johnson MD, Maglich JM, Goodwin B, Ittoop OR, Wisely B, Creech K, Parks DJ, Collins JL, Willson TM, Kalpana GV, Venkatesh M, Cho SY, Xie W, Roboz J, Redinbo M, Moore JT, Mani S (2008) The phytoestrogen coumestrol is a naturally occurring antagonist of the human pregnane X receptor. Mol Endocrinol 22:838–857

    Article  CAS  PubMed  Google Scholar 

  • Wang H, Venkatesh M, Li H, Goetz R, Mukherjee S, Biswas A, Zhu L, Kaubisch A, Wang L, Pullman J, Whitney K, Kuro-O M, Roig AI, Shay JW, Mohammadi M, Mani S (2011) Pregnane X receptor activation induces FGF19-dependent tumor aggressiveness in humans and mice. J Clin Invest 121:3220–3232

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Watkins RE, Maglich JM, Moore LB, Wisely GB, Noble SM, Davis-Searles PR, Lambert MH, Kliewer SA, Redinbo MR (2003) A crystal structure of human PXR in complex with the St John’s wort compound hyperforin. Biochemistry 42:1430–1438

    Article  CAS  PubMed  Google Scholar 

  • Xiao D, Srivastava SK, Lew KL, Zeng Y, Hershberger P, Johnson CS, Trump DL, Singh SV (2003) Allyl isothiocyanate, a constituent of cruciferous vegetables, inhibits proliferation of human prostate cancer cells by causing G2/M arrest and inducing apoptosis. Carcinogenesis 24:891–897

    Article  CAS  PubMed  Google Scholar 

  • Xie W, Barwick JL, Simon CM, Pierce AM, Safe S, Blumberg B, Guzelian PS, Evans RM (2000) Reciprocal activation of xenobiotic response genes by nuclear receptors SXR/PXR and CAR. Genes Dev 14:3014–3023

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Yun CH, Okerholm RA, Guengerich FP (1993) Oxidation of the antihistaminic drug terfenadine in human liver microsomes. Role of cytochrome P-450 3A(4) in N-dealkylation and C-hydroxylation. Drug Metab Dispos 21:403–409

    CAS  PubMed  Google Scholar 

  • Zambon S, Fontana S, Kajbaf M (2010) Evaluation of cytochrome P450 inhibition assays using human liver microsomes by a cassette analysis LC-MS/MS. Drug Metab Lett 4:120–128

    Article  CAS  PubMed  Google Scholar 

  • Zhang Y (2000) Role of glutathione in the accumulation of anticarcinogen icisothiocyanates and their glutathione conjugates by murine hepatoma cells. Carcinogenesis 21:1175–1182

    Article  CAS  PubMed  Google Scholar 

  • Zhang Y (2010) Allyl isothiocyanate as a cancer chemopreventive phytochemical. Mol Nutr Food Res 54:127–135

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Zhang Y, Talalay P (1998) Mechanism of differential potencies of isothiocyanates as inducers of anticarcinogenic Phase 2 enzymes. Cancer Res 58:4632–4639

    CAS  PubMed  Google Scholar 

  • Zhang J, Huang W, Chua SS, Wei P, Moore DD (2002) Modulation of acetaminophen-induced hepatotoxicity by the xenobiotic receptor CAR. Science 298:422–424

    Article  CAS  PubMed  Google Scholar 

  • Zhou C, Poulton EJ, Grun F, Bammler TK, Blumberg B, Thummel KE, Eaton DL (2007) The dietary isothiocyanate sulforaphane is an antagonist of the human steroid and xenobiotic nuclear receptor. Mol Pharmacol 71:220–229

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This study was supported by National Science Council, Executive Yuan, Taiwan, R.O.C. (NSC-101-2320-B-039-007-MY3), China Medical University Hospital, Taichung, Taiwan (DMR-103-049), in part by Taiwan Ministry of Health and Welfare Clinical Trial and Research Center of Excellence (MOHW103-TDU-B-212-113002), and by China Medical University under the Aim for the Top University Plan of the Ministry of Education, Taiwan. We thank Dr. Hongbing Wang (Department of Pharmaceutical Sciences, University of Maryland School of Pharmacy, Baltimore, Maryland) for kindly providing human CAR3 and CYP2B6 reporter constructs.

Conflict of interest

The authors declare that there are no conflicts of interest.

Author information

Authors and Affiliations

  1. Department of Pharmacy, College of Pharmacy, China Medical University, No. 91, Hsueh-Shih Road, Taichung, 40402, Taiwan, ROC

    Yun-Ping Lim, Ching-Hao Cheng, Wei-Cheng Chen, Dong-Zong Hung, Wei-Chih Ma, Yu-Hsien Lin & Cing-Yu Chen

  2. Department of Emergency, Toxicology Center, China Medical University Hospital, Taichung, Taiwan, ROC

    Yun-Ping Lim & Dong-Zong Hung

  3. Department of Pharmacy, Changhua Show Chwan Memorial Hospital, Changhua, Taiwan, ROC

    Ching-Hao Cheng

  4. Department of Public Health, Chung Shan Medical University, Taichung, Taiwan, ROC

    Shih-Yu Chang

  5. Graduate Institute of Pharmaceutical Technology, Tajen University, Pingtung, Taiwan, ROC

    Jih-Jung Chen

  6. School of Chinese Medicine, China Medical University, Taichung, Taiwan, ROC

    Lei Wan

  7. Drug Metabolism and Toxicology, Faculty of Pharmaceutical Sciences, Kanazawa University, Kakuma-machi, Kanazawa, Japan

    Tsuyoshi Yokoi & Miki Nakajima

  8. Graduate Institute of Integrated Medicine, China Medical University, Taichung, Taiwan, ROC

    Chao-Jung Chen

  9. Proteomics Core Laboratory, Department of Medical Research, China Medical University Hospital, Taichung, Taiwan, ROC

    Chao-Jung Chen

Authors
  1. Yun-Ping Lim
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ching-Hao Cheng
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Wei-Cheng Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Shih-Yu Chang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Dong-Zong Hung
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Jih-Jung Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Lei Wan
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Wei-Chih Ma
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Yu-Hsien Lin
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Cing-Yu Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Tsuyoshi Yokoi
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Miki Nakajima
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Chao-Jung Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Yun-Ping Lim or Chao-Jung Chen.

Electronic supplementary material

Below is the link to the electronic supplementary material.

204_2014_1230_MOESM1_ESM.pdf

Figure S1. Viability of COS-1 cells following exposure to AITC alone and in combination with rifampin, and CYP3A4 reporter transient transcription assays in COS-1 cells. (A) COS-1 cells were exposed to AITC alone (20 and 40 μM), or in combination with rifampin (20 μM), for 24 h. Cell viability was monitored as cellular acid phosphatase activity using PNPP as a substrate. The data shown are the mean ± SE (error bars) of at least triplicates. (B) COS-1 cells were co-transfected with a vector control (pcDNA3) or a PXR expression plasmid (pcDNA3-PXR), a CYP3A4 luciferase reporter plasmid, and a pRC-CMV-β-galactosidase vector. Then, the transfected cells were exposed to AITC and/or rifampin for 24 h. Luciferase activity was measured and normalized to β-galactosidase activity. The data shown are the mean ± SE (error bars) of at least triplicates. **, p < 0.01; ***, p < 0.001, vs. rifampin-treated cells. (PDF 104 kb)

204_2014_1230_MOESM2_ESM.pdf

Table S1. Effect of AITC, either alone or in combination with rifampin, on PXR, RXRα, HNF4α, CAR, SRC-1, and GR mRNA expression. HepG2 cells were treated with AITC and rifampin, either individually or in combination, for 24 h; total mRNA was collected; and the expression of the indicated genes as well as an internal control (GAPDH) was analyzed using real-time PCR. Experimental values were normalized relative to the expression of GAPDH, and expression in DMSO-treated cells was set at 1. The data shown are the mean ± SE (error bars) of triplicates. (PDF 48 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Lim, YP., Cheng, CH., Chen, WC. et al. Allyl isothiocyanate (AITC) inhibits pregnane X receptor (PXR) and constitutive androstane receptor (CAR) activation and protects against acetaminophen- and amiodarone-induced cytotoxicity. Arch Toxicol 89, 57–72 (2015). https://doi.org/10.1007/s00204-014-1230-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00204-014-1230-x

Keywords

Search

Navigation