Currently, there are no reports on the effects of enrofloxacin (EF), a fluoroquinolone antibiotic, on the cytochrome P450 enzymes in fish, although its use as antimicrobial agent in aquaculture has been put forward. Therefore, the in vivo and in vitro effects of EF on hepatic P450 enzymes of sea bass, a widespread food-producing fish, have been evaluated. Sea bass pretreated with a single dose of EF (3 mg/kg i.p.) or with three daily doses of EF (1 mg/kg i.p.) markedly depressed the microsomal N-demethylation of aminopyrine, erythromycin, the O-deethylation of 7- ethoxycoumarin, ethoxyresorufin and the 6b-testosterone hydroxylase. In vitro experiments showed that EF at 10 mM inhibited the above-mentioned activities and, in particular, the erythromycin N-demethylase (ERND) and 6btestosterone- hydroxylase, likely dependant on a P450 3A isoform. When the nature of ERND inhibition by EF was specifically studied with sea bass liver microsomes, it was found that EF is a potent mechanism-based inhibitor, with Ki of 3.7 mM and a Kinact of 0.045 min1. An immunoblot analysis with anti P450 3A27 of trout showed that the P450 3A isoform, constitutively expressed in sea bass, is particularly susceptible to inactivation by EF. In vitro experiments with sea bass microsomes have also demonstrated that EF is oxidative deethylated by the P450 system to ciprofloxacin (CF) and that this compound maintains the ability to inactivate the P450 enzymes. The mechanism by which EF or CF inactivate the P450 enzymes has not been studied but an attack of P450 on the cyclopropan ring, present, both in EF and CF structure, with the formation of electrophilic intermediates (i.e. radicals) has been postulated. In conclusion, the EF seems to be a powerful inhibitor of P450s in the sea bass. Therefore, the clinical use of this antibiotic in aquaculture has to be considered with caution.
INHIBITION OF CYTHOCROME P450 ENZYMES BY ENROFLOXACIN IN THE SEA BASS (Dicentrarchus labrax)
GIORGI, MARIO;
2003-01-01
Abstract
Currently, there are no reports on the effects of enrofloxacin (EF), a fluoroquinolone antibiotic, on the cytochrome P450 enzymes in fish, although its use as antimicrobial agent in aquaculture has been put forward. Therefore, the in vivo and in vitro effects of EF on hepatic P450 enzymes of sea bass, a widespread food-producing fish, have been evaluated. Sea bass pretreated with a single dose of EF (3 mg/kg i.p.) or with three daily doses of EF (1 mg/kg i.p.) markedly depressed the microsomal N-demethylation of aminopyrine, erythromycin, the O-deethylation of 7- ethoxycoumarin, ethoxyresorufin and the 6b-testosterone hydroxylase. In vitro experiments showed that EF at 10 mM inhibited the above-mentioned activities and, in particular, the erythromycin N-demethylase (ERND) and 6btestosterone- hydroxylase, likely dependant on a P450 3A isoform. When the nature of ERND inhibition by EF was specifically studied with sea bass liver microsomes, it was found that EF is a potent mechanism-based inhibitor, with Ki of 3.7 mM and a Kinact of 0.045 min1. An immunoblot analysis with anti P450 3A27 of trout showed that the P450 3A isoform, constitutively expressed in sea bass, is particularly susceptible to inactivation by EF. In vitro experiments with sea bass microsomes have also demonstrated that EF is oxidative deethylated by the P450 system to ciprofloxacin (CF) and that this compound maintains the ability to inactivate the P450 enzymes. The mechanism by which EF or CF inactivate the P450 enzymes has not been studied but an attack of P450 on the cyclopropan ring, present, both in EF and CF structure, with the formation of electrophilic intermediates (i.e. radicals) has been postulated. In conclusion, the EF seems to be a powerful inhibitor of P450s in the sea bass. Therefore, the clinical use of this antibiotic in aquaculture has to be considered with caution.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
