Effect of cyclophosphamide administration on the activity and relative content of hepatic P4502D1 in rat.
1. The effects of the administration of the anticancer and immunosuppressive drug, cyclophosphamide, to the rat on hepatic P4502D1 activity and content in the microsomal fraction have been examined. 2. Liver microsomes were obtained from male Hooded Wistar rats administered a single dose (i.p.) of saline or cyclophosphamide (200 mg/kg). Rats receiving cyclophosphamide were killed 1, 4, 7, 10 or 14 days after cyclophosphamide administration. The O-demethylation of dextromethorphan to dextrorphan was used to monitor 2D1 activity. 3. The mean Vmax for dextrorphan formation was reduced significantly (p < 0.0001) 7, 10 and 14 days after cyclophosphamide administration compared with the control group (control, 0.32 +/- 0.07; 7-day, 0.20 +/- 0.08; 10-day, 0.11 +/- 0.02; and 14-day group, 0.15 +/- 0.02 nmol/mg/min). 4. Western blotting revealed that there was a significant reduction (p < 0.0005) in the microsomal relative 2D1 content 10 days after cyclophosphamide administration compared with the control group (control, 1.25 +/- 0.44; and 10-day group, 0.65 +/- 0.14). 5. The activity of reduced nicotinamide adenine dinucleotide phosphate P450 reductase was significantly reduced (p < 0.0001) 7, 10 and 14 days following cyclophosphamide administration (control, 215 +/- 24; 7-day, 102 +/- 20; 10-day, 59 +/- 4 and 14-day group, 76 +/- 8 nmol/mg/min). Cytochrome b5 content was significantly reduced (p < 0.0001) 7 and 10 days following cyclophosphamide administration (control, 0.46 +/- 0.13; 7-day, 0.28 +/- 0.07 and 10-day group, 0.20 +/- 0.03 nmol/mg). 6. The significant reductions in the activity of rat hepatic microsomal 2D1 following cyclophosphamide administration, as seen by the alterations in mean Vmax for dextrorphan formation, do not appear to be due to a single factor, but may result from a combination of several events, including reductions in relative 2D1 content, reduced nicotinamide adenine dinucleotide phosphate P450-reductase activity and cytochrome b5 content.
Laslett
T J
TJ
School of Pharmacy and Medical Sciences, University of South Australia, Adelaide, Australia.
Alvarez
F
F
Nation
R L
RL
Evans
A M
AM
Scott
S D
SD
Stupans
I
I
D000818:Animals; D018906:Antineoplastic Agents, Alkylating / Q000494:pharmacology*; D015153:Blotting, Western; D002851:Chromatography, High Pressure Liquid; D003520:Cyclophosphamide / Q000494:pharmacology*; D019389:Cytochrome P-450 CYP2D6; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis / Q000378:metabolism*; D003640:Dealkylation; D003915:Dextromethorphan / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D066298:In Vitro Techniques; D007527:Isoenzymes / Q000096:biosynthesis / Q000378:metabolism*; D007700:Kinetics; D008297:Male; D008862:Microsomes, Liver / Q000187:drug effects / Q000201:enzymology* / Q000378:metabolism; D006899:Mixed Function Oxygenases / Q000096:biosynthesis / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar
D018906:Antineoplastic Agents, Alkylating; D007527:Isoenzymes; D003915:Dextromethorphan; D003520:Cyclophosphamide; D003577:Cytochrome P-450 Enzyme System; D006899:Mixed Function Oxygenases; D019389:Cytochrome P-450 CYP2D6