The use of a three compartment in vitro model to investigate the role of hepatic drug metabolism in drug-induced blood dyscrasias.
1. N-hydroxylation is thought to be an essential step in the haemotoxicity of dapsone (DDS). To investigate both metabolism-dependent and cell-selective drug toxicity in vitro we have developed a three-compartment system in which an hepatic drug metabolizing system is contained within a central compartment separated by semipermeable membranes from compartments containing mononuclear leucocytes (MNL) and red blood cells (RBC). 2. Metabolism of dapsone (100 microM) by rat liver microsomes resulted in toxicity to RBC cells (47.3 +/- 2.1% methaemoglobin), but there was no significant toxicity toward MNL (3.7 +/- 1.3% cell death) compared with control values (1.6 +/- 0.9%). However, when RBC were replaced with buffer in the third compartment there was significantly greater (P < 0.001) white cell toxicity (17.6 +/- 0.6% cell death), demonstrating the protection of MNL by RBC. Metabolism of dapsone by human liver microsomes again resulted in RBC toxicity (12.5 +/- 3.3% methaemoglobin) but no significant MNL toxicity (2.9 +/- 0.8% cell death). Replacement of RBC resulted in a significant (P < 0.001) increase in MNL toxicity (6.5 +/- 0.7% cell death). Addition of synthetic dapsone hydroxylamine (30 microM) in the absence of a metabolizing system and with no RBC in the third compartment resulted in significant (P < 0.001) toxicity toward MNL (43.36 +/- 5.82% cell death) compared with control (1.8 +/- 1.1%). The presence of RBC in the third compartment resulted in a significant (P < 0.001) decrease in MNL toxicity (17.6 +/- 2.2% cell death), with 40.1 +/- 3.7% methaemoglobin in the RBC.(ABSTRACT TRUNCATED AT 250 WORDS)
Tingle
M D
MD
Department of Pharmacology and Therapeutics, University of Liverpool.
Park
B K
BK
D000818:Animals; D003622:Dapsone / Q000378:metabolism / Q000633:toxicity; D064420:Drug-Related Side Effects and Adverse Reactions; D004906:Erythrocyte Count / Q000187:drug effects; D006402:Hematologic Diseases / Q000139:chemically induced*; D006801:Humans; D006830:Hydralazine / Q000378:metabolism / Q000633:toxicity; D066298:In Vitro Techniques; D008099:Liver / Q000378:metabolism*; D008297:Male; D008706:Methemoglobin / Q000378:metabolism; D008862:Microsomes, Liver / Q000378:metabolism; D008954:Models, Biological; D009504:Neutrophils / Q000187:drug effects; D004364:Pharmaceutical Preparations / Q000378:metabolism*; D011319:Primaquine / Q000378:metabolism / Q000633:toxicity; D011342:Procainamide / Q000378:metabolism / Q000633:toxicity; D051381:Rats; D017208:Rats, Wistar
D004364:Pharmaceutical Preparations; D006830:Hydralazine; D003622:Dapsone; D008706:Methemoglobin; D011342:Procainamide; D011319:Primaquine