Cytokine-stimulated nitric oxide production inhibits adenylyl cyclase and cAMP-dependent secretion in cholangiocytes

Carlo Spirlì, Luca Fabris, Elena Duner, Romina Fiorotto, Giorgio Ballardini, Tania Roskams, Nicholas F. Larusso, Aurelio Sonzogni, Lajos Okolicsanyi, Mario Strazzabosco

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Abstract

Background & Aims: The biliary epithelium is involved both in bile production and in the inflammatory/reparative response to liver damage. Recent data indicate that inflammatory aggression to intrahepatic bile ducts results in chronic progressive cholestasis. Methods: To understand the effects of nitric oxide on cholangiocyte secretion and biliary tract pathophysiology we have investigated: (1) the effects of proinflammatory cytokines on NO production and expression of the inducible nitric oxide synthase (NOS2), (2) the effects of NO on cAMP-dependent secretory mechanisms, and (3) the immunohistochemical expression of NOS2 in a number of human chronic liver diseases. Results: Our results show that: (1) tumor necrosis factor (TNF)-α, and interferon (IFN)-γ, synergically stimulate NO production in cultured cholangiocytes through an increase in NOS2 gene and protein expression; (2) micromolar concentrations of NO inhibit forskolin-stimulated cAMP production by adenylyl cyclase (AC), cyclic adenosine monophosphate (cAMP)-dependent fluid secretion, and cAMP-dependent Cl- and HCO3 - transport mediated by cystic fibrosis transmembrane conductance regulator (CFTR) and anion exchanger isoform 2, respectively; (3) cholestatic effects of NO and of proinflammatory cytokines are prevented by NOS-2 inhibitors and by agents (manganese(Ill)-tetrakis(4-benzoic acid)porphyrin [MnTBAP], urate, trolox) able to block the formation of reactive nitrogen oxide species (RNOS); (4) NOS2 expression is increased significantly in the biliary epithelium of patients with primary sclerosing cholangitis (PSC). Conclusions: Our findings show that proinflammatory cytokines stimulate the biliary epithelium to generate NO, via NOS2 induction, and that NO causes ductular cholestasis by a RNOS-mediated inhibition of AC and of cAMP-dependent HCO3 - and Cl- secretory mechanisms. This pathogenetic sequence may contribute to ductal cholestasis in inflammatory cholangiopathies.

Original languageEnglish (US)
Pages (from-to)737-753
Number of pages17
JournalGastroenterology
Volume124
Issue number3
DOIs
StatePublished - Mar 1 2003

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ASJC Scopus subject areas

  • Hepatology
  • Gastroenterology

Cite this

Spirlì, C., Fabris, L., Duner, E., Fiorotto, R., Ballardini, G., Roskams, T., Larusso, N. F., Sonzogni, A., Okolicsanyi, L., & Strazzabosco, M. (2003). Cytokine-stimulated nitric oxide production inhibits adenylyl cyclase and cAMP-dependent secretion in cholangiocytes. Gastroenterology, 124(3), 737-753. https://doi.org/10.1053/gast.2003.50100