Krukoski Daniel Witchmichen, Comar Samuel Ricardo, Claro Ligia Maria, Leonart Maria Suely Soares, do Nascimento Aguinaldo José
Programa de Pós-Graduação em Bioquímica e em Ciências Farmacêuticas, Universidade Federal do Paraná, Avenida Lothário Meissner 632, Jardim Botânico, Curitiba, PR, Brazil.
Hematology. 2009 Jun;14(3):168-72. doi: 10.1179/102453309X402296.
The mature human erythrocyte, when submitted to oxidative stress, can demonstrate depletion of reduced glutathione, oxidation of the hemoglobin molecule and aggregation of complexes of iron close to the membrane. These can produce abnormalities in the erythrocyte membrane and hemolysis. The aim of this work was to study the antioxidative action of vitamin C (vit. C), deferroxamine (DFO) and the flavonoids quercetin and rutin in normal human erythrocytes, submitted to in vitro oxidative stress induced by tert-butylhydroperoxide ((t)BHP). Venous blood was collected in citrate-phosphate-dextrose (CPD) solution, as anticoagulant, from healthy adult individuals after informed consent. The erythrocytes were resuspended in PBS to obtain 35% globular volume, and then submitted to the oxidative action of (t)BHP for up to 30 min, with or without previous incubation for 60 min with vit. C, DFO, quercetin and rutin. Decrease in the GSH concentration, G6-PD and GR activities, and increase in the methemoglobin and Heinz bodies (HB) formation, occurred with the increase in (t)BHP concentration. (t)BHP did not effect on the membrane proteins detected by SDS-PAGE. Quercetin, partially prevented the GSH decrease and the formation of HB, but did not prevent MetHb formation from oxidative damage by (t)BHP. Rutin, after (t)BHP induction, prevented the GSH decrease and the formation of HB. Vit. C, had no influence on the depletion of GSH, inhibited partially the metHb formation, and it protected GR, but not G6-PD from oxidative damage by (t)BHP. DFO partially inhibited the metHb formation and GSH decrease, but it did not protect GR and G6-PD from oxidative damage by (t)BHP. The results obtained suggest that vit. C, DFO and the flavonoids quercetin and rutin contribute to the decrease in the oxidative stress caused by (t)BHP.
成熟的人类红细胞在遭受氧化应激时，可表现出还原型谷胱甘肽耗竭、血红蛋白分子氧化以及靠近细胞膜的铁复合物聚集。这些情况可导致红细胞膜异常和溶血。本研究的目的是探讨维生素C（维C）、去铁胺（DFO）以及黄酮类化合物槲皮素和芦丁对正常人红细胞的抗氧化作用，这些红细胞在体外受到叔丁基过氧化氢（(t)BHP）诱导的氧化应激影响。在获得健康成年个体的知情同意后，采集静脉血并置于枸橼酸盐 - 磷酸盐 - 葡萄糖（CPD）溶液中作为抗凝剂。将红细胞重悬于磷酸盐缓冲盐水（PBS）中，使其血细胞比容达到35%，然后在有或没有预先用维C、DFO、槲皮素和芦丁孵育60分钟的情况下，使其受到(t)BHP的氧化作用长达30分钟。随着(t)BHP浓度的增加，谷胱甘肽（GSH）浓度、葡萄糖 - 6 - 磷酸脱氢酶（G6 - PD）和谷胱甘肽还原酶（GR）活性降低，高铁血红蛋白和海因茨小体（HB）形成增加。(t)BHP对通过十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳（SDS - PAGE）检测到的膜蛋白没有影响。槲皮素部分地阻止了GSH的减少和HB的形成，但没有阻止由(t)BHP引起的氧化损伤导致的高铁血红蛋白（MetHb）形成。在(t)BHP诱导后，芦丁阻止了GSH的减少和HB的形成。维C对GSH的消耗没有影响，部分抑制了MetHb的形成，并且它保护了GR，但没有保护G6 - PD免受(t)BHP的氧化损伤。DFO部分抑制了MetHb的形成和GSH的减少，但没有保护GR和G6 - PD免受(t)BHP的氧化损伤。所获得的结果表明，维C、DFO以及黄酮类化合物槲皮素和芦丁有助于降低由(t)BHP引起的氧化应激。
