PYRUVATE DEHYDROGENASE COMPLEX ACTIVITY IN THE RAT`S KIDNEYS UNDER CONDITIONS OF TOXIC ACETAMINOPHEN DAMAGE WITH PROTEIN DEFICIENCY

Abstract

The aim of this work was to study the research the enzymatic activity of pyruvate dehydrogenase complex and lac-tate dehydrogenase and determination of the lactate/pyruvate ratio in rat kidneys under conditions of toxic damage by acetaminophen against the background of protein deficiency.The activity of the pyruvate dehydrogenase complex activity was determined according to a method based on the reaction of oxidative decarboxylation of pyruvate with simultaneous reduction of NAD+, which is measured spectrophotometrically at 340 nm. LDH activity was studied by an optimized optical method, which is based on the reaction of conversion of pyruvate to lactate with concomitant oxidation of NADH, which was measured using a spectrophotometer at λ = 340 nm. Lactate content was measured spectrophotometrically by the ability of FeCl3 to interact with lactate ions, resulting in the formation of a bright yellow iron lactate solution, which is recorded at a wavelength of 390 nm. The concentration of pyruvate was deter-mined by the modified Umbright method, which is based on the reaction of the interaction of pyruvate with 2,4-Dinitrophenylhydrazine (2,4-DNPH) in an alkaline medium, during which hydrazine is formed, which was recorded using a photoelectrocolorimeter at λ = 440 nm. The study was conducted on 4 groups of experimental animals: I - control animals (K); II – rats kept on a low-protein diet (LPD); ІІІ – animals in which acute toxic damage was caused by acetaminophen (TI); IV – rats on a low-protein diet, simulated acute toxic damage with acetaminophen (LPD/TI). It is demonstrated that under the conditions of dietary protein deficiency in the kidneys, there is a decrease in the activi-ty of the pyruvate dehydrogenase complex while maintaining the control indicators of the lactate/pyruvate ratio and lactate dehydrogenase activity. However, under the conditions of the toxic effect of acetaminophen against the back-ground of dietary protein deficiency in the kidneys, an intensification of the anaerobic pathway of energy supply is observed, as evidenced by a significant decrease in the activity of the pyruvate dehydrogenase complex, an increase in the lactate/pyruvate ratio against the background of lactate dehydrogenase activation. The obtained results can be used to justify approaches to correct energy deficit in animals under the conditions of acetaminophen overdose against the background of protein deficiency.