A Method of Measuring Glutathione Peroxidase Activity in Murine Brain in Pharmacological Experiments

A method of measuring of glutathione peroxidase activity using H2O2 was adapted for homogenates of murine brains. If the amount of reduced glutathione was at the constant level of 0.55 mM, the concentration of H2O2 of 0.192 mM was saturating for glutathione peroxidase of murine brain and was selected as an optimal concentration for the estimation of enzyme activity in tris-HCl buffer with addition of NaN3 and EDTA (pH 8.5) at the incubation temperature of 37°C. The homogenates were dissolved by the reaction mixture by 10.4 times. The duration of incubation did not exceed 60 sec, if 13% homogenate was used. The experiment based on this method showed increased activity of glutathione peroxidase in the brain of mice treated with a derivative of acetaldehyde ammonia during long-term intermittent normobaric hypoxia. These data might reflect activation of glutathione peroxidase.