Peroxynitrite generated from constitutive nitric oxide synthase mediates the early biochemical injury in short‐term cultured hepatocytes

FEBS Letters

Sanz-González, Silvia M¹  López-Garcı́a, M.Pilar¹ 

[1] Departamento de Bioquı́mica y Biologı́a Molecular, Facultad de Farmacia, Universidad de Valencia, Valencia 46100, Spain

关键词: Hepatocyte isolation;    Culture;    P450 content;    Nitric oxide synthase;    Oxidative stress;    Peroxynitrite;    Protein nitration;    NO;    nitric oxide;    iNOS;    inducible nitric oxide synthase;    cNOS;    constitutive nitric oxide synthase;    L-NAME;    N G-nitro-L-arginine methyl ester;    DHR 123;    dihydrorhodamine 123;    PN;    peroxynitrite;    RH;    rhodamine;    ROS;    reactive oxygen species;    RNS;    reactive nitrogen species derived from NO;   

DOI  :  10.1016/S0014-5793(99)01789-5

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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 cultures.

【 授权许可】

Unknown