Mp. Lopez-garcia et Sm. Sanz-gonzalez, Peroxynitrite generated from constitutive nitric oxide synthase mediates the early biochemical injury in short-term cultured hepatocytes, FEBS LETTER, 466(1), 2000, pp. 187-191
Early loss of P450 in rat hepatocyte cultures appears directly related to n
itric 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-deri
ved species. NO formation at this stage is due to the early activation of l
iver constitutive NO synthase (cNOS), Immunodetection of nitrated proteins
provides direct evidence of endogenous peroxynitrite (PN) formation upon he
patocyte isolation. On the basis of the combined use of dihydrorhodamine 12
3 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 developmen
t, is the main species likely responsible for the early biochemical injury
commonly observed in hepatocyte cultures. (C) 2000 Federation of European B
iochemical Societies.