Özet:

In this study, in vitro effects of diclofenac sodium, which is used to prevent inflammation and alleviate pain in arthritis and acute injuries, on glucose-6-phosphate dehydrogenase enzyme (G6PD; EC 1.1.1.49) and glutathione reductase enzyme (GR; EC 1.8.1.7) purified from sheep spleen tissue were investigated. In the first stage of the study, G6PD and GR enzymes were purified from sheep spleen tissue by ammonium sulfate (NH4)2SO4 precipitation and 2', 5' ADP-Sepharose-4B gel affinity chromatography. The purity levels of the enzymes were checked by the sodyum dodesilsülfat poliakrilamid jel elektroforezi (SDS-PAGE) method. In the second stage of the study, the effects of diclofenac sodium on the pure enzyme activities were investigated. IC50 values were calculated by plotting the % Activity-[I] graph for diclofenac sodium, which has an inhibitory effect on enzymes. In addition, Lineweaver-Burk graphs were drawn by making measurements with five different substrate concentrations and three different inhibitor concentrations. From these graphs, it was determined that G6PD enzyme inhibited with 0.51 mM IC50 value and 0.48 ± 0.01 mM Ki value non competitively, by diclofenac sodium. It was determined that the GR enzyme inhibited with an 6.55 mM IC50 value of and 15.69 ± 0.84 mM Ki value non competitively.