Identification of HMG-CoA reductase inhibitors as activators for human, mouse and rat constitutive androstane receptor.
Constitutive active (or androstane) receptor (CAR, NR1I3), a member of the nuclear receptor family, is a major regulator for induction of cytochrome P450 2B (CYP2B) genes by phenobarbital. Phenobarbital-like inducer, 1,4-bis[2-(3,5-dichloropyridyloxy)]benzene is a potent mouse CAR ligand that has been used to study CAR target genes in mice but does not activate human CAR (hCAR) or rat CAR (rCAR). Although 6-(4-chlorophenyl) imidazo[2,1-b][1,3]thiazole-5-carbaldehyde O-(3,4-dichlorobenzyl)oxime (CITCO) was reported to be an hCAR agonistic ligand, activation of hCAR by CITCO in cell-based reporter assay was weak. Therefore, we performed a screening of 50 drugs and chemicals using cell-based reporter assays to identify activators of hCAR. Among them, HMG-CoA reductase inhibitors (cerivastatin, simvastatin, fluvastatin, and atorvastatin) enhanced the hCAR-mediated transcriptional activation of phenobarbital-responsive enhancer module reporter gene by up to 3-fold. Similar activation by HMG-CoA reductase inhibitors was also observed with mouse and rat CARs. On the other hand, pravastatin did not activate hCAR at the concentrations tested (up to 30 microM). The extent of activation by the HMG-CoA reductase inhibitors was stronger than that by CITCO. Cerivastatin, simvastatin, fluvastatin, and atorvastatin induced CYP2B6 mRNA in stable hCAR-expressed FLC7 cells but not in original FLC7 cells. Therefore, we concluded that CAR mediates the effects of HMG-CoA reductase inhibitors on the induction of CYP2B genes, although HMG-CoA reductase inhibitors also activate pregnane X receptor. HMG-CoA reductase inhibitors such as cerivastatin would be useful to study for elucidating molecular and cellular mechanisms of hCAR.
Kobayashi
Kaoru
K
Laboratory of Pharmacology and Toxicology, Graduate School of Pharmaceutical Sciences, Chiba University, Chuo-ku, Chiba 260-8675, Japan. kaoruk@p.chiba-u.ac.jp
Yamanaka
Yosuke
Y
Iwazaki
Norihiko
N
Nakajo
Ikumi
I
Hosokawa
Masakiyo
M
Negishi
Masahiko
M
Chiba
Kan
K
D000728:Androgens; D000731:Androstanes / Q000378:metabolism*; D000818:Animals; D001483:Base Sequence; D045744:Cell Line, Tumor; D000090702:Constitutive Androstane Receptor; D017931:DNA Primers; D017930:Genes, Reporter; D006801:Humans; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors / Q000494:pharmacology*; D051379:Mice; D011725:Pyridines / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D011944:Receptors, Androgen / Q000235:genetics / Q000378:metabolism; D015533:Transcriptional Activation
D000728:Androgens; D000731:Androstanes; D000090702:Constitutive Androstane Receptor; D017931:DNA Primers; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors; C000716327:NR1I3 protein, human; C000716328:Nr1i3 protein, mouse; C000716329:Nr1i3 protein, rat; D011725:Pyridines; D012333:RNA, Messenger; D011944:Receptors, Androgen; C086276:cerivastatin; C033936:androstane