title: Metabolism and cytotoxicity of propyl gallate in isolated rat hepatocytes: effects of a thiol reductant and an esterase inhibitor.
abstract: The relationship between the metabolism and the cytotoxic effects of propyl gallate (PG) has been studied in freshly isolated rat hepatocytes. Addition of PG (0.5-2.0 mM) to the hepatocytes elicited concentration-dependent cell death, accompanied by decreases in intracellular ATP, adenine nucleotide pools, glutathione, and protein thiols. The rapid loss of ATP preceded the onset of cell death. PG in the hepatocyte suspensions was converted to gallic acid, 4-O-methyl-gallic acid, and other minor products over time. In addition, PG was converted to a dimer [dipropyl-4,4',5,5',6,6'-hexahydroxydiphenate (PG-dimer)] and ellagic acid via autooxidation. In comparisons of the toxic effects of PG and its metabolites at concentrations of 2 mM, the parent compound PG was the most toxic. Pretreatment of hepatocytes with diazinon (100 microM), an esterase inhibitor, enhanced PG-induced cytotoxicity. This was accompanied by delay of PG loss and inhibition of gallic acid formation. The cytotoxicity of PG was also enhanced by addition of the thiol reductant dithiothreitol (4 mM), although intracellular levels of glutathione and protein thiols were maintained during the incubation period. Dithiothreitol did not affect the hydrolysis of PG to gallic acid by esterases but did delay the conversion of PG and prevented the formation of PG-dimer. In isolated hepatic mitochondria, PG elicited a concentration-dependent increase in the rate of state 4 oxygen consumption, indicating an uncoupling effect. In contrast, PG-dimer inhibited the rate of state 3 oxygen consumption. Based on the respiratory control index, the order of potency for impairment of mitochondria was PG > PG-dimer > gallic acid = 4-O-methyl-gallic acid = ellagic acid - propyl alcohol. These results indicate (a) that PG-induced hepatotoxicity is mediated by the parent compound and not its metabolites, (b) that toxicity is associated with ATP depletion apparently independently of cellular thiol depletion, and (c) that mitochondria may represent critical targets of PG-induced cytotoxicity.
authors.affiliation: Department of Toxicology, Tokyo Metropolitan Research Laboratory of Public Health, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Nakagawa
authors.forename: K
authors.initials: K
authors.lastname: Nakajima
authors.forename: S
authors.initials: S
authors.lastname: Tayama
authors.forename: P
authors.initials: P
authors.lastname: Moldéus
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D016923:Cell Death / Q000187:drug effects; D003976:Diazinon / Q000494:pharmacology; D004229:Dithiothreitol / Q000494:pharmacology; D004950:Esterases / Q000037:antagonists & inhibitors; D005978:Glutathione / Q000378:metabolism; D066298:In Vitro Techniques; D008099:Liver / Q000187:drug effects* / Q000378:metabolism*; D008297:Male; D008930:Mitochondria, Liver / Q000187:drug effects / Q000378:metabolism; D010101:Oxygen Consumption / Q000187:drug effects; D011435:Propyl Gallate / Q000737:chemistry / Q000378:metabolism* / Q000633:toxicity*; D051381:Rats; D011916:Rats, Inbred F344
chemical_list: D011435:Propyl Gallate; D000255:Adenosine Triphosphate; D004950:Esterases; D005978:Glutathione; D004229:Dithiothreitol; D003976:Diazinon