Determination of the free radical reactions in whole blood of Wistar rats under stress-induced physical exercise

Purpose. Determination of the free radical reactions in whole blood of Wistar rats under stress-induced physical exercise. Materials and methods. The object of the research is whole blood of male Wistar rats. Rats in the experimental group were forced to swim in an aquarium (swimming time 40 minutes at the water temperature of 25 °C with using of Porsolt test). The swimming time was determined experimentally after the stress modeling. The values of maximum luminescence intensity (Imax) and its growth rate (L tg) were determined with spontaneous luminol-dependent chemiluminescence on BCLM 3606M biochemiluminometer. The rate of the free radical reaction was calculated by using the formula described by Yu.A. Vladimirov et al. (2011). Results. During stress-induced physical exercise Imax value significantly increases by 1.09 times (p < 0.05) and L tg value significantly decreases by 4.65 times (p < 0.05). Between the above-noted parameters the linear dependence described with the equation Imax = 128.411L tg where 128.411 is the coefficient characterizing BCLM 3606M analyzer's sensitivity to radiation of a photomultiplier tube is observed. The rate constants of the studied free radical reactions were established with this equation: the constant of the first reaction k1 = 11010 (mol/dm3)-1·s-1 is the constant of peroxynitrite formation rate, the constant of the second reaction k2 = 3.2107 (mol/dm3)-1·s-1 is the constant of the rate of reduction of glutathione-thiyl radical to reduced form of glutathione. Conclusions. As a result of our study, it was found that there is the reaction of formation of peroxynitrite from radical of nitrogen monoxide and the reaction of reduction of glutathione-thiyl radical to reduced form of glutathione during stress-induced physical exercise. For the first time free radical reactions during stress-induced physical exercise were determined with the dependence of luminescence rate on ratio of maximum chemiluminescence intensity to the coefficient characterizing BCLM 3606M analyzer sensitivity to radiation of a photomultiplier tube. For the first time the equation described by Yu.A. Vladimirov et al. (2011) used in study of free radical reactions using BCLM 360* analyzers. In the process of chemiluminescence substances are not destroyed, therefore a reaction path doesn't change. In addition, chemiluminescence is highly sensitive, which is very important when registering highly reactive radicals.