Up-regulation of CYP1A1 by rutaecarpine is dependent on aryl hydrocarbon receptor and calcium

doi:10.1016/j.tox.2009.10.013

Toxicology

Volume 266, Issues 1–3, 21 December 2009, Pages 38-47

https://doi.org/10.1016/j.tox.2009.10.013 Get rights and content

Abstract

Rutaecarpine is a quinazolinocarboline alkaloid isolated from a traditional Chinese medicinal fruit, Evodia rutaecarpa. In the present study, we investigated the effect of rutaecarpine on CYP1A1 expression mediated by [Ca²⁺] and the AhR pathway in mouse hepatoma Hepa-1c1c7 cells. Rutaecarpine also significantly increased CYP1A1 enzyme activity and mRNA and protein levels. Rutaecarpine markedly induced XRE and AhR binding activity. CH-223191, an AhR antagonist, blocked the rutaecarpine-induced CYP1A1 enzyme activity and mRNA and protein expression. In addition, rutaecarpine remarkably induced the phosphorylation of Ca²⁺/calmodulin (CaM)-dependent protein kinase (CaMK). W7 and BAPTA/AM, a CaM antagonist and an intracellular Ca²⁺ chelator, respectively, blocked the rutaecarpine-induced CYP1A1 enzyme activity and mRNA and protein expression. These results indicate that rutaecarpine induces CYP1A1 expression through AhR- and calcium-dependent mechanisms.

Introduction

Evodia fruit is an herbal medicine prepared from the matured fruit of the plant Evodia rutaecarpa (Iwata et al., 2005). Rutaecarpine, a major component of Evodia fruit, has been used in traditional medicine to treat gastrointestinal disorders, headache, and hypertension, and as an anti-inflammatory agent (Moon et al., 1999, Woo et al., 2001, Takada et al., 2005, Ding et al., 2008, Li et al., 2008, Liu et al., 2008). The effect of rutaecarpine can be blocked by capsazepine, a transient receptor potential vanilloid subtype 1 (TRPV1) antagonist, suggesting that TRPV1 plays a key role in these protective effects of rutaecarpine (Ueng et al., 2001); TRPV1 is a ligand-gated nonselective cation channel (Ding et al., 2008). Previously, rutaecarpine was shown to induce CYP1A1 in mouse and rat liver microsomes (Ueng et al., 2001, Ueng et al., 2002a, Lee et al., 2004b) and to act as a potent inhibitor of CYP1A2 in mouse and human liver microsomes (Ueng et al., 2002b). In addition, Gillner et al. (1989) demonstrated binding of rutaecarpine to the rat liver cytosolic AhR. However, the mechanism of CYP1A1 regulation by rutaecarpine has not been fully characterized. Recently, rutaecarpine was found to induce the activity and expression of Phase 2 carcinogen detoxifying enzymes, such as NAD(P)H:quinone reductase (QR) in Hepa-1c1c7 cells (Ahn et al., 2008).

The aryl hydrocarbon receptor (AhR) is a member of the basic helix-loop-helix DNA binding protein family (Whitlock, 1999) and can be activated by structurally diverse synthetic and natural chemicals. AhR mediates the toxic and biological effects of polycyclic aromatic hydrocarbons (PAH) such as 3-methylcholanthrene (3-MC) (Jeuken et al., 2003). CYP1A1 oxidatively biotransforms various PAH and has been used to investigate the mechanisms of aryl hydrocarbon (Ah) actions. The control and regulation of CYP1A1 gene expression have been investigated extensively. It is currently believed that CYP1A1 gene expression is induced by 3-MC and related AhR agonists through Ah receptor-mediated signal transduction. Upon binding with ligand, AhR forms a heterodimer with AhR nuclear translocator (ARNT) and binds to specific DNA recognition sequences known as xenobiotic-response elements (XREs) located upstream of the CYP1A1 transcription start site (Delescluse et al., 2000, Mimura and Fujii-Kuriyama, 2003).

In addition, rutaecarpine increases intracellular [Ca²⁺] levels in cultured rat endothelial cells and vascular smooth muscle cells (Whitlock, 1999). Activation of AhR increases the intracellular concentration of Ca²⁺ (Tannheimer et al., 1997, Le Ferrec et al., 2002, Barouki et al., 2007, N’Diaye et al., 2006). [Ca²⁺]/calmodulin (CaM)-dependent protein kinase (CaMK) is a member of a family of structurally related serine/threonine protein kinases that play important roles in proliferation (Rodriguez-Mora et al., 2005) and differentiation (Zayzafoon, 2006). However, CYP1A1 and calcium interaction has not been fully characterized.

In this study, we demonstrate that rutaecarpine can stimulate the expression of CYP1A1 through AhR- and calcium-dependent mechanisms.

Section snippets

Materials

All chemicals and cell culture materials were obtained from the following sources: Rutaecarpine (purity, >99.8%), 3-MC and capsazepine from Sigma Chemical Co. (St. Louis, MO, USA); 2-methyl-2H-pyrazole-3-carboxylic acid (2-methyl-4-o-tolylazo-phenyl)-amide (CH-223191); 1,2-bis(O-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid tetra (acetoxymethyl) ester (BAPTA-AM) and CaMK inhibitor W7 (N-(6-Aminohexyl)-5-chloro-1-naphthalenesulfonamide) from Calbiochem (La Jolla, CA, USA); Fluo-3 AM from

CYP1A1 levels are increased in rutaecarpine-treated Hepa-1c1c7 cells

To determine the optimal concentrations to use in our studies, the potential cytotoxicity of rutaecarpine was tested in Hepa-1c1c7 and Tao BpRc1 cells. The chemical structure of rutaecarpine is shown in Fig. 1A. Fig. 1B shows that 0.01, 0.1, 1 and 10 μM rutaecarpine did not affect cell viability; however, 100 μM, the highest concentration tested, caused a 38% decrease in cell viability. Therefore, all subsequent studies were conducted using 0.01–1 μM rutaecarpine.

CYP1A1 enzyme activity was

Discussion

Rutaecarpine, from Evodia fruit induces the activity and expression of Phase 2 carcinogen detoxifying enzymes, such as NAD(P)H:quinone reductase (QR) in Hepa-1c1c7 cells (Ahn et al., 2008). Previous studies suggest that rutaecarpine has a chemopreventive effect through its, ability to modulate carcinogen detoxification pathways. However, regulation of Phase 1 enzymes (CYP 450), which catalyze the initial step of xenobiotic detoxification, by rutaecarpine and its molecular mechanism have not

Conflict of interest statement

None declared.

Acknowledgement

This work was supported by grants of Priority Research Centers Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (2009-1813) and National Institute of Toxicological Research (07142KFDA570) by Korea Food and Drug Administration (KFDA).

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Up-regulation of CYP1A1 by rutaecarpine is dependent on aryl hydrocarbon receptor and calcium

Abstract

Introduction

Section snippets

Materials

CYP1A1 levels are increased in rutaecarpine-treated Hepa-1c1c7 cells

Discussion

Conflict of interest statement

Acknowledgement

FEBS Lett.

Int. J. Immunopharmacol.

Food Chem. Toxicol.

Trends Cell Biol.

Neuropharmacology

Toxicology

J. Biol. Chem.

Anal. Biochem.

Cell

Drug Metab. Pharmacokinet.

Food Chem. Toxicol.

Biochem. Biophys. Res. Commun.

Anal. Biochem.

Toxicology

Biochem. Biophys. Res. Commun.

J. Biol. Chem.

J. Biol. Chem.

Biochim. Biophys. Acta.

J. Biol. Chem.

J. Biol. Chem.

Jpn. J. Pharmacol.

Life Sci.

Induction of NAD(P)H: quinone reductase by rutaecarpine isolated from the fruits of Evodia rutaecarpa in the murine hepatic Hepa-1c1c7 cell line

Planta Med.

7,12-Dimethylbenz[a]anthracene activates protein-tyrosine kinases Fyn and Lck in the HPB-ALL human T-cell line and increases tyrosine phosphorylation of phospholipase C-gamma 1, formation of inositol 1,4,5-trisphosphate, and mobilization of intracellular calcium

Proc. Natl. Acad. Sci. U.S.A.

Aromatic hydrocarbon responsiveness-receptor agonists generated from indole-3-carbinol in vitro and in vivo: comparisons with 2,3,7,8-tetrachlorodibenzo-p-dioxin

Proc. Natl. Acad. Sci. U.S.A.