title: Biliary secretion of S-nitrosoglutathione is involved in the hypercholeresis induced by ursodeoxycholic acid in the normal rat.
abstract: Ursodeoxycholic acid (UDCA) induces bicarbonate-rich hypercholeresis by incompletely defined mechanisms that involve the stimulation of adenosine triphosphate (ATP) release from cholangiocytes. As nitric oxide (NO) at a low concentration can stimulate a variety of secretory processes, we investigated whether this mediator could be implicated in the choleretic response to UDCA. Our in vivo experiments with the in situ perfused rat liver model in anesthetized rats, showed that UDCA infusion increased the biliary secretion of NO derivatives, hepatic inducible NO synthase expression, and NO synthase activity in liver tissue. UDCA also stimulated NO release by isolated rat hepatocytes. In contrast to UDCA, cholic acid was a poor inducer of NO secretion, and tauroursodeoxycholic acid showed no effect on NO secretion. Upon UDCA administration, NO was found in bile as low-molecular-weight nitrosothiols, of which S-nitrosoglutathione (GSNO) was the predominant species. UDCA-stimulated biliary NO secretion was abolished by the inhibition of inducible NO synthase with N(omega)-nitro-L-arginine methyl ester in isolated perfused livers and also in rats whose livers were depleted of glutathione with buthionine sulfoximine. Moreover, the biliary secretion of NO species was significantly diminished in UDCA-infused transport mutant [ATP-binding cassette C2 (ABCC2)/multidrug resistance-associated protein 2 (Mrp2)-deficient] rats, and this finding was consistent with the involvement of the glutathione carrier ABCC2/Mrp2 in the canalicular transport of GSNO. It was particularly noteworthy that in cultured normal rat cholangiocytes, GSNO activated protein kinase B, protected against apoptosis, and enhanced UDCA-induced ATP release to the medium; this effect was blocked by phosphoinositide 3-kinase inhibition. Finally, retrograde GSNO infusion into the common bile duct increased bile flow and biliary bicarbonate secretion.
  
  
  CONCLUSIONS
  UDCA induces biliary secretion of GSNO, which contributes to stimulating ductal secretion.
authors.affiliation: Division of Hepatology and Gene Therapy, CIMA University of Navarra, Pamplona, Spain. jprieto@unav.es
authors.forename: Carlos M
authors.initials: CM
authors.lastname: Rodríguez-Ortigosa
authors.forename: Jesús M
authors.initials: JM
authors.lastname: Banales
authors.forename: Israel
authors.initials: I
authors.lastname: Olivas
authors.forename: Iker
authors.initials: I
authors.lastname: Uriarte
authors.forename: José J G
authors.initials: JJ
authors.lastname: Marín
authors.forename: Fernando J
authors.initials: FJ
authors.lastname: Corrales
authors.forename: Juan F
authors.initials: JF
authors.lastname: Medina
authors.forename: Jesús
authors.initials: J
authors.lastname: Prieto
mesh_terms: D000818:Animals; D001646:Bile / Q000187:drug effects / Q000378:metabolism*; D002478:Cells, Cultured; D022781:Hepatocytes; D008099:Liver / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D009569:Nitric Oxide / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D026422:S-Nitrosoglutathione / Q000378:metabolism*; D026403:S-Nitrosothiols / Q000378:metabolism; D014580:Ursodeoxycholic Acid / Q000494:pharmacology
chemical_list: D026403:S-Nitrosothiols; D009569:Nitric Oxide; D026422:S-Nitrosoglutathione; D014580:Ursodeoxycholic Acid