title: PGE1 protection against apoptosis induced by D-galactosamine is not related to the modulation of intracellular free radical production in primary culture of rat hepatocytes.
abstract: D-galactosamine (D-GalN) toxicity is a useful experimental model of liver failure in human. It has been previously observed that PGE1 treatment reduced necrosis and apoptosis induced by D-GalN in rats. Primary cultured rat hepatocytes were used to evaluate if intracellular oxidative stress was involved during the induction of apoptosis and necrosis by D-GalN (0-40mM). Also, the present study investigated if PGE1 (1 microM) was equally potent reducing both types of cell death. The presence of hypodiploid cells, DNA fragmentation and caspase-3 activation were used as a marker of hepatocyte apoptosis. Necrosis was measured by lactate dehydrogenase (LDH) release. Oxidative stress was evaluated by the intracellular production of hydrogen peroxide (H2O2), the disturbances on the mitochondrial transmembrane potential (MTP), thiobarbituric-reacting substances (TBARS) release and the GSH/GSSG ratio. Data showed that intermediate range of D-GalN concentrations (2.5-10mM) induced apoptosis in association with a moderate oxidative stress. High D-GalN concentration (40 mM) induced a reduction of all parameters associated with apoptosis and enhanced all those related to necrosis and intracellular oxidative stress, including a reduction of GSH/GSSG ratio and MTP in comparison with D-GalN (2.5-10 mM)-treated cells. Although PGE1 reduced apoptosis induced by D-GalN, it was not able to reduce the oxidative stress and cell necrosis induced by the hepatotoxin in spite to its ability to abolish the GSH depletion.
authors.affiliation: Unidad de Investigación, Servicio Aparato Digestivo, Hospital Universitario Reína Sofía, Córdoba, Spain.
authors.forename: A
authors.initials: A
authors.lastname: Quintero
authors.forename: C A
authors.initials: CA
authors.lastname: Pedraza
authors.forename: E
authors.initials: E
authors.lastname: Siendones
authors.forename: A M
authors.initials: AM
authors.lastname: Kamal ElSaid
authors.forename: A
authors.initials: A
authors.lastname: Colell
authors.forename: C
authors.initials: C
authors.lastname: García-Ruiz
authors.forename: J L
authors.initials: JL
authors.lastname: Montero
authors.forename: M
authors.initials: M
authors.lastname: De la Mata
authors.forename: J C
authors.initials: JC
authors.lastname: Fernández-Checa
authors.forename: G
authors.initials: G
authors.lastname: Miño
authors.forename: J
authors.initials: J
authors.lastname: Muntané
mesh_terms: D000527:Alprostadil / Q000494:pharmacology*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D056486:Chemical and Drug Induced Liver Injury / Q000473:pathology* / Q000517:prevention & control; D053938:DNA Fragmentation; D005434:Flow Cytometry; D005609:Free Radicals; D005688:Galactosamine; D005978:Glutathione / Q000378:metabolism; D022781:Hepatocytes / Q000187:drug effects* / Q000473:pathology; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D015227:Lipid Peroxidation; D008099:Liver / Q000187:drug effects* / Q000473:pathology; D008297:Male; D008564:Membrane Potentials; D008928:Mitochondria / Q000378:metabolism; D009336:Necrosis; D010975:Platelet Aggregation Inhibitors / Q000494:pharmacology*; D011003:Ploidies; D051381:Rats; D017208:Rats, Wistar; D017382:Reactive Oxygen Species / Q000378:metabolism; D017392:Thiobarbituric Acid Reactive Substances / Q000378:metabolism
chemical_list: D005609:Free Radicals; D010975:Platelet Aggregation Inhibitors; D017382:Reactive Oxygen Species; D017392:Thiobarbituric Acid Reactive Substances; D005688:Galactosamine; D007770:L-Lactate Dehydrogenase; C505407:CASP3 protein, human; C505410:Casp3 protein, rat; D053148:Caspase 3; D020169:Caspases; D000527:Alprostadil; D005978:Glutathione