Defenses against oxidation in human erythrocytes: role of glutathione reductase in the activation of glucose decarboxylation by hemolytic drugs.
We have used 1,3-bis(2-chloroethyl)-1-nitrosourea, a selective inhibitor of oxidized glutathione reductase (GSSG-R), to examine the role of this enzyme in regulating the hexose monophosphate shunt (HMS) and to explore how a variety of agents influence glucose decarboxylation in intact human red blood cells (RBCs). Substances tested included primaquine and several other drugs that are specially hemolytic and methemoglobinemic in glucose-6-phosphate dehydrogenase (G6PD) deficiency and related disorders. The results allowed us to distinguish and quantitate contrasting modes of HMS stimulation and to clarify how RBCs respond to different classes of oxidants. Some agents like methylene blue (MB), phenazine methosulfate, and pyrroline carboxylate do not require GSSG-R to increase CO2 production; they activate G6PD and 6-phosphogluconic dehydrogenase by directly oxidizing reduced nicotinamide adenine dinucleotide phosphate (NADPH) to oxidized nicotinamide adenine dinucleotide phosphate (NADP). Other compounds, like ascorbate, nitrofurantoin, and doxorubicin, oxidize GSH primarily; CO2 increases indirectly only when GSSG-R, activated by glutathione disulfide (GSSG), raises the level of NADP. Chemicals like primaquine, daunorubicin, and methylphenylazoformate trigger the HMS by independently oxidizing both NADPH and GSH. Unlike MB, most drugs that are hemolytic in G6PD deficiency activate the HMS in a manner that depends to a variable extent on GSSG-R. This variability may explain hitherto puzzling clinical and pharmacogenetic differences between primaquine and diaminodiphenylsulfone-induced hemolysis.
Hohl
R J
RJ
Department of Pharmacology (Genetics) and Medicine (Hematology), Rush-Presbyterian-St. Luke's Medical Center, Rush University, Chicago, Illinois 60612.
Kennedy
E J
EJ
Frischer
H
H
D002245:Carbon Dioxide / Q000378:metabolism; D002330:Carmustine / Q000494:pharmacology; D003653:Decarboxylation; D004912:Erythrocytes / Q000378:metabolism*; D005947:Glucose / Q000378:metabolism*; D005955:Glucosephosphate Dehydrogenase Deficiency / Q000097:blood; D005980:Glutathione Reductase / Q000378:metabolism*; D006461:Hemolysis / Q000187:drug effects*; D006801:Humans; D009249:NADP / Q000378:metabolism; D010084:Oxidation-Reduction; D010427:Pentose Phosphate Pathway
D002245:Carbon Dioxide; D009249:NADP; D005980:Glutathione Reductase; D005947:Glucose; D002330:Carmustine