Farnesoid X receptor and bile salts are involved in... : Hepatology (original) (raw)

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Farnesoid X receptor and bile salts are involved in transcriptional regulation of the gene encoding the human bile salt export pump

Plass, Jacqueline R. M.1; Mol, Olaf1; Heegsma, Janette1; Geuken, Mariska1; Faber, Klaas Nico*,1; Jansen, Peter L. M.1; Müller, Michael1, 2

1 Center for Liver, Digestive, and Metabolic Diseases, Groningen, The Netherlands

2 University Wageningen, Nutrition, Metabolism, and Genomics Group, Bomenweg 2, 6703 HD Wageningen, The Netherlands. O.M.'s current address is: DSM Biologics, Zuiderweg 72-2 9744 AP Groningen, The Netherlands.

E-mail:[email protected]

*Address reprint requests to: Department of Gastroenterology and Hepatology, University Hospital Groningen, P.O. Box 30.001, 9700 RB Groningen, The Netherlands. fax: (31) 50-3614756.

Received April 18, 2001; accepted December 12, 2001; previously published online December 30, 2003

Abstract

The bile salt export pump (BSEP or ABCB11) mediates the adenosine triphosphate-dependent transport of bile salts across the canalicular membrane of the hepatocyte. Mutations in the corresponding ABCB11 gene cause progressive familial intrahepatic cholestasis type 2. The aim of this study was to investigate the regulation of human ABCB11 gene transcription by bile salts. First, a 1.7-kilobase human ABCB11 promoter region was cloned. Sequence analysis for possible regulatory elements showed a farnesoid X receptor responsive element (FXRE) at position −180. The farnesoid X receptor (FXR) functions as a heterodimer with the retinoid X receptor α (RXRα) and can be activated by the bile salt chenodeoxycholic acid (CDCA). Luciferase reporter gene assays showed that the ABCB11 promoter is positively controlled by FXR, RXRα, and bile salts in a concentration-dependent manner. Mutation of the FXRE strongly represses the FXR-dependent induction. Second, endogenous ABCB11 transcription regulation was studied in HepG2 cells, stably expressing the rat sodium-dependent taurocholate transporter (rNtcp) cells. ABCB11 expression was induced by adding bile salts to the culture medium, and this effect was maximized by combining it with cotransfection of rFxr and hRXRα. Reducing endogenous FXR levels using RNA interference fully repressed the bile salt-induced ABCB11 expression. In conclusion, these results show that FXR is required for the bile salt-dependent transcriptional control of the human ABCB11 gene and that the cellular amount of FXR is critical for the level of activation of ABCB11 transcription.