Useful agents for the study of glutathione metabolism in erythroyctes. Organic hydroperoxides.
Useful agents for the study of glutathione metabolism in erythroyctes. Organic hydroperoxides.
复制标题
用于研究红细胞谷胱甘肽代谢的有用试剂。
作者：
Samir K. Srivastava;Yogesh C. Awasthi;Ernest Beutler
叔丁基过氧化氢和枯烯过氧化氢，都是已知的谷胱甘肽过氧化物酶的底物，用于氧化红细胞GSH。在红细胞或全血中加入相对于GSH等摩尔浓度的氢过氧化物，在37 ℃下以4.5s的半衰期定量氧化红细胞中的GSH，在4 ℃下约为3倍。在葡萄糖存在下，正常红细胞在约25分钟内再生所有GSH。然而，葡萄糖6-磷酸脱氢酶缺陷的红细胞不能再生GSH。用氢过氧化物处理红细胞不影响兔红细胞存活。用等摩尔浓度的氢过氧化物氧化红细胞GSH不会导致血红蛋白和GSH的混合二硫化物的形成。过氧化氢不影响红细胞糖酵解和己糖一磷酸分流途径酶。先前关于GSSG从红细胞转运的研究通过使用叔丁基过氧化氢氧化红细胞GSH得到证实。
t-Butyl hydroperoxide and cumene hydroperoxide, both known to be substrates for glutathione peroxidase, were used to oxidize erythrocyte GSH. Addition of concentrations of hydroperoxides equimolar with respect to GSH in the erythrocytes or whole blood quantitatively oxidizes GSH in the erythrocytes with a half-time of 4.5s at 37 degrees C and about three times as long at 4 degrees C. In the presence of glucose, normal erythrocytes regenerate all the GSH in about 25min. However, glucose 6-phosphate dehydrogenase-deficient erythrocytes failed to regenerate GSH. Treatment of erythrocytes with hydroperoxides does not affect erythrocyte survival in rabbits. Oxidation of erythrocyte GSH with equimolar concentrations of hydroperoxides does not lead to formation of mixed disulphides of haemoglobin and GSH. The hydroperoxides do not affect erythrocyte glycolytic and hexose monophosphate-shunt-pathway enzymes. Previous studies on transport of GSSG from erythrocytes were confirmed by using t-butyl hydroperoxide to oxidize erythrocyte GSH.