Role of NF-kappaB activation and nitric oxide expression during PGE protection against d-galactosamine-induced cell death in cultured rat hepatocytes

Liver Int. 2004 Jun;24(3):227-36. doi: 10.1111/j.1478-3231.2004.00913.x.

Abstract

Prostaglandin E1 (PGE1) reduces cell death in experimental and clinical liver dysfunction. Nitric oxide (NO) mediates PGE1 protection against D-galactosamine (D-GalN)-induced cell death. Nuclear factor kappa-B (NF-kappaB) plays a protective role in different experimental models of cell death. We investigated if NF-kappaB was responsible for inducible nitric oxide synthase (iNOS) expression and cytoprotection induced by PGE1 against D-GalN cell death in cultured hepatocytes. Rat hepatocytes were isolated following the classical method of collagenase perfusion of liver. A kinetic study of cell death, NF-kappaB activation, mRNA and protein iNOS expression, and NO production was carried in hepatocytes treated with D-GalN (5 mM) in the presence or absence of PGE1 (1 microM) administered 2 h before the hepatotoxin. A proteasome inhibitor was used to evaluate the role of NF-kappaB activation in our experimental conditions. PGE1 protection against D-GalN-induced cell death was associated with its capacity to rapidly enhance NF-kappaB activation, mRNA and protein iNOS expression, and NO production in D-GalN-treated hepatocytes. The inhibition of NF-kappaB activation abolished iNOS expression and cell protection by PGE1 in hepatocytes treated with the hepatotoxin. The present study shows that the cytoprotection by PGE1 against D-GalN-induced apoptosis was related to NF-kappaB-dependent iNOS expression.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alprostadil / pharmacology*
  • Animals
  • Apoptosis
  • Cell Death
  • Cells, Cultured
  • Cysteine Endopeptidases
  • Cytoprotection
  • Enzyme Induction / physiology
  • Galactosamine / pharmacology*
  • Hepatocytes / drug effects*
  • Hepatocytes / physiology*
  • I-kappa B Proteins / metabolism
  • Male
  • Multienzyme Complexes / antagonists & inhibitors
  • NF-KappaB Inhibitor alpha
  • NF-kappa B / physiology*
  • Necrosis
  • Nitric Oxide / metabolism*
  • Nitric Oxide Synthase / biosynthesis
  • Nitric Oxide Synthase / genetics
  • Nitric Oxide Synthase Type II
  • Proteasome Endopeptidase Complex
  • RNA, Messenger / metabolism
  • Rats
  • Rats, Wistar

Substances

  • I-kappa B Proteins
  • Multienzyme Complexes
  • NF-kappa B
  • Nfkbia protein, rat
  • RNA, Messenger
  • NF-KappaB Inhibitor alpha
  • Nitric Oxide
  • Galactosamine
  • Nitric Oxide Synthase
  • Nitric Oxide Synthase Type II
  • Nos2 protein, rat
  • Cysteine Endopeptidases
  • Proteasome Endopeptidase Complex
  • Alprostadil