title: Cyclosporin A-sensitive cytotoxicity of flurbiprofen non-stereoselectively mediated by cytochrome P450 metabolism in three-dimensional cultured rat hepatocytes.
abstract: OBJECTIVE
  2-Arylpropionic acid (profen) drugs are associated with severe hepatotoxicity; however, risk factors are still poorly understood. Acyl-coenzyme A (acyl-CoA) thioesters of profen drugs play a more important role in the covalent binding to rat hepatocyte proteins than the respective acyl-glucuronides. Therefore, we examined whether acyl-glucuronides, acyl-CoA thioesters and oxidative metabolites of profen drugs stereoselectively participated in liver damage.
  
  
  METHODS
  Cytotoxicity was determined by measuring lactate dehydrogenase (LDH) leakage from three-dimensional cultured rat hepatocytes.
  
  
  RESULTS
  LDH leakage was not induced by R-2-phenylpropionic acid and R-ibuprofen greatly forming acyl-CoA thioesters. S-Naproxen metabolized mainly by Uridine 5'-diphosphate (UDP)-glucuronosyl-transferase did not enhance LDH leakage. However, flurbiprofen (FLP) induced LDH leakage. A selective cytochrome P450 (CYP) 2C11 inhibitor suppressed 40-50% of the R-FLP and S-FLP-induced cytotoxicity. Borneol non-stereoselectively accelerated the FLP-induced cytotoxicity. The R-FLP-induced cytotoxicity decreased intracellular adenosine triphosphate (ATP) levels to 50% of untreated hepatocytes. An inhibitor of mitochondrial permeability transition pore, cyclosporin A (Cys A), rescued ATP levels and LDH leakage back to control levels.
  
  
  CONCLUSIONS
  The reactive acyl-CoA thioesters and acyl-glucuronides were not associated with liver damage, denying one of the leading hypotheses. CYP metabolism of FLP non-stereoselectively participated in Cys A-sensitive cytotoxicity, suggesting mitochondrial injury.
authors.affiliation: Faculty of Pharmaceutical Sciences, Tokushima Bunri University, Tokushima, Japan.
authors.forename: Tadatoshi
authors.initials: T
authors.lastname: Tanino
authors.affiliation: Faculty of Pharmacy, Kinki University, Higashi-Osaka, Osaka, Japan.
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Funakami
authors.affiliation: Faculty of Pharmacy, Kinki University, Higashi-Osaka, Osaka, Japan.
authors.forename: Noriaki
authors.initials: N
authors.lastname: Nagai
authors.affiliation: Kagawa School of Pharmaceutical Sciences, Tokushima Bunri University, Sanuki, Kagawa, Japan.
authors.forename: Yoshihisa
authors.initials: Y
authors.lastname: Kato
mesh_terms: D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D056486:Chemical and Drug Induced Liver Injury / Q000209:etiology / Q000473:pathology; D016572:Cyclosporine / Q000494:pharmacology*; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D005480:Flurbiprofen / Q000633:toxicity*; D022781:Hepatocytes / Q000187:drug effects* / Q000378:metabolism; D007052:Ibuprofen / Q000633:toxicity; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008297:Male; D008928:Mitochondria / Q000473:pathology; D009288:Naproxen / Q000633:toxicity; D010666:Phenylpropionates / Q000633:toxicity; D051381:Rats; D017208:Rats, Wistar; D013237:Stereoisomerism
chemical_list: D010666:Phenylpropionates; D009288:Naproxen; D005480:Flurbiprofen; D016572:Cyclosporine; D000255:Adenosine Triphosphate; D003577:Cytochrome P-450 Enzyme System; C013025:hydratropic acid; D007770:L-Lactate Dehydrogenase; D007052:Ibuprofen
keywords: cytochrome P450-catalysed oxidation; cytotoxicity; mitochondrial permeability transition pore opening; three-dimensional cultured rat hepatocytes