title: Cytotoxicity of monocrotaline in isolated rat hepatocytes: effects of dithiothreitol and fructose.
abstract: Monocrotaline (MCT) is a pyrrolizidine alkaloid present in plants of the Crotalaria species that causes cytotoxicity and genotoxicity, including hepatotoxicity in animals and humans. It is metabolized by cytochrome P-450 in the liver to the alkylating agent dehydromonocrotaline (DHM). In previous studies using isolated rat liver mitochondria, we observed that DHM, but not MCT, inhibited the activity of respiratory chain complex I and stimulated the mitochondrial permeability transition with the consequent release of cytochrome c. In this study, we evaluated the effects of MCT and DHM on isolated rat hepatocytes. DHM, but not MCT, caused inhibition of the NADH-linked mitochondrial respiration. When hepatocytes of rats pre-treated with dexamethasone were incubated with MCT (5 mM), they showed ALT leakage, impaired ATP production and decreased levels of intracellular reduced glutathione and protein thiols. In addition, MCT caused cellular death by apoptosis. The addition of fructose or dithiotreitol to the isolated rat hepatocyte suspension containing MCT prevented the ATP depletion and/or glutathione or thiol oxidation and decreased the ALT leakage and apoptosis. These results suggest that the toxic effect of MCT on hepatocytes may be caused by metabolite-induced mitochondrial energetic impairment, together with a decrease of cellular glutathione and protein thiols.
authors.affiliation: Laboratório de Bioquímica Metabólica e Toxicológica, UNESP-Univ Estadual Paulista, Campus de Dracena, 17900-000 Dracena, SP, Brazil.
authors.forename: Marcos A
authors.initials: MA
authors.lastname: Maioli
authors.forename: Larissa C
authors.initials: LC
authors.lastname: Alves
authors.forename: Diego
authors.initials: D
authors.lastname: Perandin
authors.forename: Andréa F
authors.initials: AF
authors.lastname: Garcia
authors.forename: Flávia T V
authors.initials: FT
authors.lastname: Pereira
authors.forename: Fábio E
authors.initials: FE
authors.lastname: Mingatto
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D029864:Crotalaria / Q000378:metabolism; D004229:Dithiothreitol / Q000494:pharmacology*; D005632:Fructose / Q000494:pharmacology*; D005978:Glutathione / Q000378:metabolism; D022781:Hepatocytes / Q000187:drug effects* / Q000378:metabolism; D008297:Male; D008930:Mitochondria, Liver / Q000187:drug effects* / Q000378:metabolism; D016686:Monocrotaline / Q000031:analogs & derivatives / Q000633:toxicity*; D011506:Proteins / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D013438:Sulfhydryl Compounds / Q000378:metabolism
chemical_list: D011506:Proteins; D013438:Sulfhydryl Compounds; C014114:monocrotaline pyrrole; D005632:Fructose; D016686:Monocrotaline; D000255:Adenosine Triphosphate; D005978:Glutathione; D004229:Dithiothreitol