Impaired endothelial nitric oxide synthase activity associated with enhanced caveolin binding in experimental cirrhosis in the rat
A reduction in nitric oxide (NO) has been implicated as a cause of intrahepatic vasoconstriction in cirrhosis, but the regulatory mechanisms remain undefined. The aim of this study was to examine a contributory role for caveolin-1, a putative negative regulator of endothelial NO synthase, in mediati...
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Published in | Gastroenterology (New York, N.Y. 1943) Vol. 117; no. 5; p. 1222 |
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Main Authors | , , , , , , , |
Format | Journal Article |
Language | English |
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United States
01.11.1999
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Abstract | A reduction in nitric oxide (NO) has been implicated as a cause of intrahepatic vasoconstriction in cirrhosis, but the regulatory mechanisms remain undefined. The aim of this study was to examine a contributory role for caveolin-1, a putative negative regulator of endothelial NO synthase, in mediating deficient intrahepatic NO production in the intact cirrhotic liver.
Cirrhosis was induced by carbon tetrachloride inhalation. Flow regulation of NO production and perfusion pressure was examined in the perfused rat liver. Protein expression of endothelial NO synthase (eNOS), caveolin, and calmodulin was examined by Western blotting and immunohistochemistry. NOS activity and NO production were assessed by citrulline generation and chemiluminescence, respectively. Protein-protein interactions were examined using whole tissue protein immunoprecipitation.
In response to incremental increases in flow, cirrhotic animals produced significantly less NO(x) than control animals. NOS activity was significantly reduced in liver tissue from cirrhotic animals compared with control animals in the presence of similar eNOS protein levels. Deficient eNOS activity was associated with a severalfold increase in binding of eNOS with caveolin. Protein levels of caveolin-1 were markedly increased in the cirrhotic liver.
These studies provide evidence that enhanced expression and interaction of caveolin with eNOS contribute to impaired NO production, reduced NOS activity, and vasoconstriction in the intact cirrhotic liver. |
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AbstractList | A reduction in nitric oxide (NO) has been implicated as a cause of intrahepatic vasoconstriction in cirrhosis, but the regulatory mechanisms remain undefined. The aim of this study was to examine a contributory role for caveolin-1, a putative negative regulator of endothelial NO synthase, in mediating deficient intrahepatic NO production in the intact cirrhotic liver.
Cirrhosis was induced by carbon tetrachloride inhalation. Flow regulation of NO production and perfusion pressure was examined in the perfused rat liver. Protein expression of endothelial NO synthase (eNOS), caveolin, and calmodulin was examined by Western blotting and immunohistochemistry. NOS activity and NO production were assessed by citrulline generation and chemiluminescence, respectively. Protein-protein interactions were examined using whole tissue protein immunoprecipitation.
In response to incremental increases in flow, cirrhotic animals produced significantly less NO(x) than control animals. NOS activity was significantly reduced in liver tissue from cirrhotic animals compared with control animals in the presence of similar eNOS protein levels. Deficient eNOS activity was associated with a severalfold increase in binding of eNOS with caveolin. Protein levels of caveolin-1 were markedly increased in the cirrhotic liver.
These studies provide evidence that enhanced expression and interaction of caveolin with eNOS contribute to impaired NO production, reduced NOS activity, and vasoconstriction in the intact cirrhotic liver. |
Author | Papapetropoulos, A Toruner, M Shah, V Sessa, W C Choo, K Groszmann, R J Haddad, F Cadelina, G |
Author_xml | – sequence: 1 givenname: V surname: Shah fullname: Shah, V organization: Department of Medicine, Yale University School of Medicine, New Haven, Connecticut, USA – sequence: 2 givenname: M surname: Toruner fullname: Toruner, M – sequence: 3 givenname: F surname: Haddad fullname: Haddad, F – sequence: 4 givenname: G surname: Cadelina fullname: Cadelina, G – sequence: 5 givenname: A surname: Papapetropoulos fullname: Papapetropoulos, A – sequence: 6 givenname: K surname: Choo fullname: Choo, K – sequence: 7 givenname: W C surname: Sessa fullname: Sessa, W C – sequence: 8 givenname: R J surname: Groszmann fullname: Groszmann, R J |
BackLink | https://www.ncbi.nlm.nih.gov/pubmed/10535886$$D View this record in MEDLINE/PubMed |
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SubjectTerms | Animals Calmodulin - metabolism Caveolin 1 Caveolins In Vitro Techniques Liver - enzymology Liver Circulation - physiology Liver Cirrhosis, Experimental - metabolism Male Membrane Proteins - metabolism Nitric Oxide - biosynthesis Nitric Oxide Synthase - antagonists & inhibitors Nitric Oxide Synthase - metabolism Nitric Oxide Synthase Type III Perfusion Pressure Rats Rats, Sprague-Dawley Reference Values Tissue Distribution |
Title | Impaired endothelial nitric oxide synthase activity associated with enhanced caveolin binding in experimental cirrhosis in the rat |
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