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

Abstract

A method of measuring of glutathione peroxidase activity using HO 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 HO 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 NaN 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.

Hydrogen Sulfide Alleviates Anxiety, Motor, and Cognitive Dysfunctions in Rats with Maternal Hyperhomocysteinemia via Mitigation of Oxidative Stress

“…The antioxidant potential was determined by measuring the activity of glutathione peroxidase (GPx) assessed by the decrease of the reduced form of glutathione (GSH) level using tert-butyl as a substrate [ 54 ]. One mL of glutathione solution was mixed with 1 mL of brain homogenate; the mixture was divided into two centrifuge tubes (test and control) and incubated for 5 min.…”

Protective Effects of Probiotics on Cognitive and Motor Functions, Anxiety Level, Visceral Sensitivity, Oxidative Stress and Microbiota in Mice with Antibiotic-Induced Dysbiosis

“…The antioxidant potential was determined by measuring the activity of glutathione peroxidases (GPx) using tert-butyl hydroperoxide as a substrate according to the method described by Razygraev et al (2018) [48].…”

Investigation of oxidative processes and revealing the changes in purine nucleoside phosphorylase activity levels in model animals in the development of diabetes mellitus

“…Glutathione in complex with glutathione peroxidase and glutathione reductase makes a system which has an important role in the regulation of metabolic processes in cells, the main function of these enzymes is the reduction of hydroperoxides to alcohols [15].…”

“…Mitochondrial fraction was isolated in a medium containing 0,25 M saccharose and 0, 01M tris buffer, pH - 7,4, by a method of differential centrifugation on a K-24 centrifuge. Glutathione peroxidase activity was assessed by reduction reactions of hydrogen peroxide (H2O2) and lipid hydroperoxides (ROOH) in the presence of GSH [12, 15].…”