Oxygen-dependent hepatotoxicity due to doxorubicin: role of reducing equivalent supply in perfused rat liver.
Doxorubicin is an important anticancer drug that undergoes redox cycling leading to the production of oxygen radicals; however, its clinical use is limited by toxicity. Redox cycling due to doxorubicin was assessed in the perfused rat liver from increases in O2 uptake by the organ, and toxicity was determined from lactate dehydrogenase release and trypan blue uptake. Doxorubicin increased O2 uptake in a concentration-related manner with half-maximal increases at about 100 microM drug. Within 5 min after addition of 300 microM doxorubicin, lactate dehydrogenase was detected in the effluent perfusate. Enzyme release increased steadily and reached values of 600 units/liter after 60 min. Rates of O2 uptake due to redox cycling of doxorubicin (300 microM) increased by 57 mumol/g/hr in oxygen-rich (mean [O2] = 473 microM) periportal regions of the liver lobule, but did not change in pericentral regions where O2 tension was lower [( O2] = 247 microM). Concomitantly, fluorescence of NAD(P)H measured from the liver surface decreased in periportal but not pericentral regions. The zone-specific decrease in NADPH was attributed to redox cycling of doxorubicin. Trypan blue was taken up exclusively by cells in periportal regions of the liver lobule after perfusion with doxorubicin. When the average O2 tension was lowered from 550 to 200 microM, O2 uptake due to redox cycling of doxorubicin in periportal regions was reduced 3-fold and toxicity was abolished, indicating that toxicity due to doxorubicin is oxygen-dependent. Redox cycling of doxorubicin was minimal in regions of the perfused liver where the O2 concentration was below 400 microM. In contrast, isolated microsomes displayed maximal changes in O2 uptake due to redox cycling of doxorubicin at O2 tensions of about 10 microM. Thus, oxygen per se is not rate-limiting for redox cycling of doxorubicin in the intact organ. Since NADPH is also required for redox cycling of doxorubicin, the effect of oxygen on the ability of mitochondria and the pentose cycle to supply reducing equivalents for redox cycling of doxorubicin was examined. NADPH supply from the pentose cycle was reduced by fasting while that from mitochondria was inhibited by cyanide. The increase in O2 uptake due to redox cycling of doxorubicin was around 60 mumol/g/hr in livers from fed or fasted rats. In the presence of potassium cyanide, stimulation of O2 uptake by doxorubicin was reduced by about one-half in livers from fed rats (29 mumol/g/hr) yet was abolished nearly completely in livers from fasted rats (7 mumol/g/hr).(ABSTRACT TRUNCATED AT 400 WORDS)
Ganey
P E
PE
Department of Pharmacology, University of North Carolina, Chapel Hill 27599.
Kauffman
F C
FC
Thurman
R G
RG
D000818:Animals; D004317:Doxorubicin / Q000378:metabolism / Q000633:toxicity*; D005260:Female; D008099:Liver / Q000187:drug effects*; D009249:NADP / Q000032:analysis / Q000494:pharmacology; D010084:Oxidation-Reduction; D010100:Oxygen / Q000032:analysis / Q000494:pharmacology*; D010477:Perfusion; D011190:Potassium Cyanide / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains
D009249:NADP; D004317:Doxorubicin; D011190:Potassium Cyanide; D010100:Oxygen