Effects of some analgesic anaesthetic drugs on human erythrocyte glutathione reductase: anin vitrostudy

Abstract

Inhibitory effects of some analgesic and anaesthetic drugs on human erythrocyte glutathione reductase were investigated. For this purpose, human erythrocyte glutathione reductase was initially purified 2139-fold in a yield of 29% by using 2 0 , 5 0 -ADP Sepharose 4B affinity gel and Sephadex G-200 gel filtration chromatography. SDS polyacrylamide gel electrophoresis confirmed the purity of the enzyme by sharing a single band. A constant temperature (þ 48C) was maintained during the purification process. Diclofenac sodium, ketoprofen, lornoxicam, tenoxicam, etomidate, morphine and propofol exhibited inhibitory effects on the enzyme in vitro using the Beutler assay method.

Susceptibility of cord blood antioxidant enzymes glutathione reductase, glutathione peroxidase and glutathione S-transferase to different antibiotics:in vitroapproach

“…These enzymes are cooperative in several aspects 32,33 . Our groups recently investigated the interaction of human erythrocyte GR enzyme with several types of analgesic, anaesthetic drugs and antibiotics as well as metal ions 5,[9][10][11][12][13][14][15] .…”

“…The blood samples were centrifuged at 2250g for 15 min and the plasma and buffy coat were removed. The packed red cells were washed three times with KCl (0.16 M) and hemolyzed with 5 volume of ice-cold water and then centrifuged (4°C, 10,000g, for 30 min) to remove the ghosts and intact cells 5,11 Ammonium sulphate precipitation…”

“…Active fractions were collected and dialyzed with equilibration buffer. All of the procedures were performed at 4°C 5,13 . After dried Sephadex G-200 (2 g) was used for a 165 ml column (2 × 50 cm) bed volume.…”

“…In each fraction, enzyme activity was determined at 340 nm. Active fractions were collected and stored at −20°C for testing the enzyme purity by electrophoresis 5,9,11,15 .…”

“…Glutathione reductase (EC 1.8.1.7, GR) is an ubiquitous flavoenzyme that catalyzes the NAD(P)H-dependent reduction of GSSG to GSH. This enzyme maintains adequate levels of the cellular GSH pool, which is critical not only for maintaining the cellular redox status by keeping sulfhydryl groups of cytosolic proteins in their reduced form 4,5 but also because numerous toxic or potentially toxic compounds, including some metals, are either taken up by or research arTIcLe Susceptibility of cord blood antioxidant enzymes glutathione reductase, glutathione peroxidase and glutathione S-transferase to different antibiotics: in vitro approach removed from the cells by GSH-mediated pathways 6 . Glutathione peroxidase (EC 1.11.1.9, GPx) catalyzes the reduction of hydrogen peroxide by GSH.…”