Peroxynitrite generated from constitutive nitric oxide synthase mediates the early biochemical injury in short‐term cultured hepatocytes
- 18 January 2000
- journal article
- Published by Wiley in FEBS Letters
- Vol. 466 (1), 187-191
- https://doi.org/10.1016/s0014-5793(99)01789-5
Abstract
Early loss of P450 in rat hepatocyte cultures appears directly related to nitric oxide (NO) overproduction. This study provides experimental evidence for the induction – shortly after isolation through the classical procedure – of strong oxidative stress that involves both oxygen-derived and NO-derived species. NO formation at this stage is due to the early activation of liver constitutive NO synthase (cNOS). Immunodetection of nitrated proteins provides direct evidence of endogenous peroxynitrite (PN) formation upon hepatocyte isolation. On the basis of the combined use of dihydrorhodamine 123 and NOS inhibitors, the analysis of the amount, time course and nature of the species involved supports the view that PN generated from cNOS-derived NO, while not affecting cell viability and hepatocyte monolayer development, is the main species likely responsible for the early biochemical injury commonly observed in hepatocyte culturesKeywords
This publication has 26 references indexed in Scilit:
- Formation and Loss of Nitrated Proteins in Peroxynitrite-Treated Rat Skin in VivoBiochemical and Biophysical Research Communications, 1999
- Peroxynitrite Induces Tyrosine Nitration and Modulates Tyrosine Phosphorylation of Synaptic ProteinsJournal of Neurochemistry, 1999
- Rapid Induction of NF-κB Binding during Liver Cell Isolation and Culture: Inhibition byL-NAMEIndicates a Role for Nitric Oxide SynthaseBiochemical and Biophysical Research Communications, 1999
- Endogenous nitric oxide is responsible for the early loss of P450 in cultured rat hepatocytesFEBS Letters, 1998
- Scavenging of hydroxyl radicals but not of peroxynitrite by inhibitors and substrates of nitric oxide synthasesBritish Journal of Pharmacology, 1997
- Decreased constitutive hepatic nitric oxide synthase expression in secondary biliary fibrosis and its changes after Roux-en-Y choledocho-jejunostomy in the ratJournal of Hepatology, 1996
- Peroxynitrite formed by simultaneous generation of nitric oxide and superoxide selectively inhibits bovine aortic prostacyclin synthaseFEBS Letters, 1996
- Peroxynitrite-mediated oxidation of dihydrorhodamine 123Free Radical Biology & Medicine, 1994
- A Fluorometric Assay for the Measurement of Nitrite in Biological SamplesAnalytical Biochemistry, 1993
- Peroxynitrite-mediated tyrosine nitration catalyzed by superoxide dismutaseArchives of Biochemistry and Biophysics, 1992