IN VITRO INVESTIGATION OF THE INHIBITORY EFFECT OF HEPATOTOXIC DIETHYLNITROSAMINE COMPOUND ON XANTHINE OXIDASE ENZYME

Abstract

Aim: It was aimed to examine the inhibition effect of diethyl nitrosamine compound, one of the nitrosole compounds known to have carcinogenic and hepato-toxic effects, on xanthine oxidase enzyme in vitro. Materials-Method: For this purpose, the inhibition effect of the diethyl nitrosamine compound prepared at 3 different concentrations (2mM, 4mM, 6mM) on the xanthine oxidase enzyme in the substrate medium at 4 different concentrations (0.3 mg/mL, 0.5 mg/mL, 1 mg/mL, 2 mg/mL) was examined. Xanthine oxidase enzyme activity was determined spectrophotometrically by reading the absorbance of urate formed from hypoxanthine at a wavelength of 290 nm according to the Worthington. Michaelis Menten and Lineweaver Burk graphs were drawn with the data obtained. Km, Vmax values of the enzyme were calculated with the help of the equation obtained from the drawn Lineweaver Burk plot. Additionally, the IC50 values of the enzyme were calculated with the help of graphics. GraphPad Prism 6 program was used for drawing graphs and calculations. Results: It was observed that while the Vmax values obtained as a result of the calculations decreased, the Km values remained constant. In the light of the Lineweaver Burk plot and calculations, it was concluded that the diethyl nitrosamine compound has a non-competitive inhibition on the xanthine oxidase enzyme. Conclusion: We found that the diethyl nitrosamine compound non-competitively inhibits the xanthine oxidase enzyme, which is known to play important roles in many biological processes. The compound diethyl nitrosamine, which is known to have hepatotoxic and carcinogenic effects even at low doses, binds reversibly to the structure of the enzyme, but is very harmful in terms of side effects. It is not possible to use it for therapeutic purposes, and exposure to this compound may cause worse pathophysiological processes by inhibiting many enzymes and pathways, including xanthine oxidase