Bile-acid-activated farnesoid X receptor regulates hydrogen sulfide production and hepatic microcirculation.
OBJECTIVE
To investigate whether the farnesoid X receptor (FXR) regulates expression of liver cystathionase (CSE), a gene involved in hydrogen sulfide (H(2)S) generation.


METHODS
The regulation of CSE expression in response to FXR ligands was evaluated in HepG2 cells and in wild-type and FXR null mice treated with 6-ethyl chenodeoxycholic acid (6E-CDCA), a synthetic FXR ligand. The analysis demonstrated an FXR responsive element in the 5'-flanking region of the human CSE gene. The function of this site was investigated by luciferase reporter assays, chromatin immunoprecipitation and electrophoretic mobility shift assays. Livers obtained from rats treated with carbon tetrachloride alone, or in combination with 6-ethyl chenodeoxycholic acid, were studied for hydrogen sulphide generation and portal pressure measurement.


RESULTS
Liver expression of CSE is regulated by bile acids by means of an FXR-mediated mechanism. Western blotting, qualitative and quantitative polymerase chain reaction, as well as immunohistochemical analysis, showed that expression of CSE in HepG2 cells and in mice is induced by treatment with an FXR ligand. Administration of 6E-CDCA to carbon tetrachloride treated rats protected against the down-regulation of CSE expression, increased H(2)S generation, reduced portal pressure and attenuated the endothelial dysfunction of isolated and perfused cirrhotic rat livers.


CONCLUSIONS
These results demonstrate that CSE is an FXR-regulated gene and provide a new molecular explanation for the pathophysiology of portal hypertension.
Department of Clinical and Experimental Medicine University of Perugia, Via E dal Pozzo, 06122 Perugia, Italy. barbara.renga@unipg.it
Barbara
B
Renga
Andrea
A
Mencarelli
Marco
M
Migliorati
Eleonora
E
Distrutti
Stefano
S
Fiorucci
D000316:Adrenergic alpha-Agonists / Q000494:pharmacology; D000818:Animals; D001483:Base Sequence; D001647:Bile Acids and Salts / Q000378:metabolism*; D002251:Carbon Tetrachloride / Q000494:pharmacology; D002460:Cell Line; D003542:Cystathionine gamma-Lyase / Q000235:genetics / Q000378:metabolism; D005355:Fibrosis / Q000139:chemically induced; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D006862:Hydrogen Sulfide / Q000378:metabolism*; D006975:Hypertension, Portal / Q000378:metabolism; D008099:Liver / Q000098:blood supply* / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D051379:Mice; D018345:Mice, Knockout; D008833:Microcirculation; D008969:Molecular Sequence Data; D009638:Norepinephrine / Q000494:pharmacology; D011401:Promoter Regions, Genetic; D051381:Rats; D018160:Receptors, Cytoplasmic and Nuclear / Q000235:genetics / Q000378:metabolism*; D000099150:Receptor, Farnesoid X-Activated
D000316:Adrenergic alpha-Agonists; D001647:Bile Acids and Salts; D002251:Carbon Tetrachloride; D003542:Cystathionine gamma-Lyase; D006862:Hydrogen Sulfide; D009638:Norepinephrine; D018160:Receptors, Cytoplasmic and Nuclear; D000099150:Receptor, Farnesoid X-Activated