Impact of cephaloridine on glutathione and related enzymes: comparison of in vivo and in vitro rat models.
The aim of this study was to investigate the early effects of cephaloridine (CPH) on glutathione-dependent phase II detoxification in the rat proximal tubular cell and to find an in vitro alternative to the in vivo model. The in vivo study was conducted in three groups of rats which received CPH at doses of 250, 500 or 750 mg/kg per day for 3 days, while another group received 500 mg/kg as a single dose. For the in vitro study, rat renal proximal tubular cultured cells were exposed to CPH at concentrations of 0.3, 0.6, 1, 1.7 mM for 24, 48 and 72 h. Glutathione-dependent detoxification was evaluated in vivo and in vitro on the basis of total intracellular glutathione (GSH), glutathione S-transferase (GST) and glutathione peroxidase (GPX). Glutathione reductase (GRED) and GST mRNA levels were also determined. Results of in vivo and in vitro models were comparable in terms of the early increase of GSH, GST and GRED. This increase had a bell-shaped dose-response with a maximum at 500 mg/kg in vivo and 1 mM in vitro. Beyond these doses, GSH and its dependent enzyme levels decreased, associated with cytotoxicity in vitro and renal insufficiency in vivo. The increased GST activity was associated with an increased level of GST7 in vivo and a markedly increased level of GST1-2 in vitro. We concluded that the in vitro model can be used as an alternative to animal experimentation to study glutathione-dependent detoxication. Low cytotoxic doses of CPH induced an early increase of glutathione phase II-dependent detoxification enzymes.
Moritz
F
F
Intensive Care Unit, Charles Nicolle University Medical Center, Rouen, France.
Marouillat
S
S
Monteil
C
C
Baudelot
A
A
Fillastre
J P
JP
Bonmarchand
G
G
Morin
J P
JP
D000818:Animals; D001711:Biotransformation / Q000187:drug effects; D002478:Cells, Cultured; D002509:Cephaloridine / Q000633:toxicity*; D005260:Female; D005978:Glutathione / Q000187:drug effects* / Q000378:metabolism*; D005982:Glutathione Transferase / Q000187:drug effects / Q000378:metabolism; D007687:Kidney Tubules, Proximal / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D008954:Models, Biological; D051381:Rats; D017208:Rats, Wistar
D005982:Glutathione Transferase; D005978:Glutathione; D002509:Cephaloridine