The role of reactive oxygen species in adriamycin and menadione-induced glomerular toxicity.
Redox cycling leading to oxidative stress has been proposed as the mechanism by which adriamycin induces glomerular toxicity in rats. The present study compares the extent of the oxidative stress and cytotoxicity induced by adriamycin to menadione (a model redox cycling quinone) in freshly isolated rat glomeruli. Adriamycin and menadione (25 microM) decreased de novo protein synthesis (measured by 3H-proline incorporation into acid-precipitable glomerular protein) by 50 and 85%, respectively, in 2 h. By contrast, menadione at 25 microM reduce glomerular membrane integrity (as assessed by lactate dehydrogenase leakage), adriamycin reduced membrane integrity at 500 microM adriamycin. Reactive oxygen species (ROS) were measured by the oxidation of dihydrodichlorofluorescein. Menadione (25 microM) and adriamycin (25 microM) increased ROS formation to 260 and 156% of controls after 30 min incubation, respectively. Oxidative stress was assessed by measuring the intracellular level of reduced glutathione (GSH) and the decrease of the NADPH/NADP- ratio which stimulates the pentose phosphate pathway (PPP): (a) menadione (25-100 microM) reduced glomerular GSH to 10-20% of controls, adriamycin (25-100 microM) had no effect; (b) menadione (10 microM) increased PPP activity 6-fold, while adriamycin (125 microM) had only a 2-fold effect. Although adriamycin and menadione generate extensive ROS and decrease protein synthesis, there was no correlation between the extent of oxidative stress and cytotoxicity in glomeruli exposed to adriamycin. These results suggest that oxidative stress may not be the primary mechanisms by which adriamycin induces selective glomerular toxicity.
Morgan
W A
WA
Department of Life Sciences, Faculty of Science and Health, University of East London, UK. W.A.Morgan@UEL.AC.UK
Kaler
B
B
Bach
P H
PH
D000818:Animals; D000903:Antibiotics, Antineoplastic / Q000633:toxicity*; D004317:Doxorubicin / Q000633:toxicity*; D005978:Glutathione / Q000032:analysis; D006490:Hemostatics / Q000633:toxicity*; D007674:Kidney Diseases / Q000139:chemically induced*; D007678:Kidney Glomerulus / Q000187:drug effects* / Q000473:pathology; D007770:L-Lactate Dehydrogenase / Q000032:analysis / Q000378:metabolism; D008297:Male; D008560:Membrane Fluidity / Q000187:drug effects; D018384:Oxidative Stress; D010427:Pentose Phosphate Pathway / Q000187:drug effects; D014176:Protein Biosynthesis; D051381:Rats; D017208:Rats, Wistar; D017382:Reactive Oxygen Species / Q000378:metabolism*; D014812:Vitamin K / Q000633:toxicity*
D000903:Antibiotics, Antineoplastic; D006490:Hemostatics; D017382:Reactive Oxygen Species; D014812:Vitamin K; D004317:Doxorubicin; D007770:L-Lactate Dehydrogenase; D005978:Glutathione