Evaluation of a method for determinationof glutathionereductase activity inerythrocytes
Glutathione (GSH) is a molecule that consists of three amino acids: glutamic acid, cysteine and glycine. GSH has several important functions: to protect cells from free radicals, reactive oxygen species and oxidative stress. GSH exists in a reduced form, GSH, and in an oxidized dimeric form, glutationdisulfid, GSSG. The enzymes glutathionereductase (GR) catalyses the reduction of GSSG back to GSH. Nicotinamide adenine dinucleotide phosphate (NADPH) is required as a coenzyme in the reaction. Deficiency of GSH in the body results in lysis of red blood cells leading to reduced life length of these cells and hemolytic anemia. In this study,we evaluated a method for determining the GR activity used in the investigation of hemolytic anemia. Blood samples containing both normal and high reticulocyte concentration were washed, to extract red blood cells. To analyze GR activity, we used an enzyme activity assay based on spectrofotometry. The method showed reliable results for GR activity and hopefully will be used in diagnostic routines.