title: The nuclear receptor FXR regulates hepatic transport and metabolism of glutamine and glutamate.
abstract: Hepatic transport and metabolism of glutamate and glutamine are regulated by intervention of several proteins. Glutamine is taken up by periportal hepatocytes and is the major source of ammonia for urea synthesis and glutamate for N-acetylglutamate (NAG) synthesis, which is catalyzed by the N-acetylglutamate synthase (NAGS). Glutamate is taken up by perivenous hepatocytes and is the main source for the synthesis of glutamine, catalyzed by glutamine synthase (GS). Accumulation of glutamate and ammonia is a common feature of chronic liver failure, but mechanism that leads to failure of the urea cycle in this setting is unknown. The Farnesoid X Receptor (FXR) is a bile acid sensor in hepatocytes. Here, we have investigated its role in the regulation of the metabolism of both glutamine and glutamate. In vitro studies in primary cultures of hepatocytes from wild type and FXR(-/-) mice and HepG2 cells, and in vivo studies, in FXR(-/-) mice as well as in a rodent model of hepatic liver failure induced by carbon tetrachloride (CCl(4)), demonstrate a role for FXR in regulating this metabolism. Further on, promoter analysis studies demonstrate that both human and mouse NAGS promoters contain a putative FXRE, an ER8 sequence. EMSA, ChIP and luciferase experiments carried out to investigate the functionality of this sequence demonstrate that FXR is essential to induce the expression of NAGS. In conclusion, FXR activation regulates glutamine and glutamate metabolism and FXR ligands might have utility in the treatment of hyperammonemia states.
authors.affiliation: Dipartimento di Medicina Clinica e Sperimentale, Università degli Studi di Perugia, Italy. barbara.renga@unipg.it
authors.forename: Barbara
authors.initials: B
authors.lastname: Renga
authors.forename: Andrea
authors.initials: A
authors.lastname: Mencarelli
authors.forename: Sabrina
authors.initials: S
authors.lastname: Cipriani
authors.forename: Claudio
authors.initials: C
authors.lastname: D'Amore
authors.forename: Angela
authors.initials: A
authors.lastname: Zampella
authors.forename: Maria Chiara
authors.initials: MC
authors.lastname: Monti
authors.forename: Eleonora
authors.initials: E
authors.lastname: Distrutti
authors.forename: Stefano
authors.initials: S
authors.lastname: Fiorucci
mesh_terms: D051046:Amino-Acid N-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D000818:Animals; D015153:Blotting, Western; D002251:Carbon Tetrachloride / Q000633:toxicity; D002467:Cell Nucleus; D047369:Chromatin Immunoprecipitation; D024202:Electrophoretic Mobility Shift Assay; D005971:Glutamates / Q000378:metabolism*; D005973:Glutamine / Q000378:metabolism*; D056945:Hep G2 Cells; D022781:Hepatocytes / Q000166:cytology / Q000378:metabolism; D006801:Humans; D022124:Hyperammonemia / Q000139:chemically induced / Q000378:metabolism*; D008106:Liver Cirrhosis, Experimental / Q000139:chemically induced / Q000378:metabolism*; D008156:Luciferases / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D011401:Promoter Regions, Genetic / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D018160:Receptors, Cytoplasmic and Nuclear / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000099150:Receptor, Farnesoid X-Activated
chemical_list: D051046:Amino-Acid N-Acetyltransferase; D002251:Carbon Tetrachloride; D005971:Glutamates; D005973:Glutamine; D008156:Luciferases; D012333:RNA, Messenger; D018160:Receptors, Cytoplasmic and Nuclear; D000099150:Receptor, Farnesoid X-Activated; C494874:NAGS protein, human; C494877:Nags protein, mouse; C016195:N-acetylglutamic acid