Genistein abrogates pre-hemolytic and oxidative stress damage induced by 2,2'-Azobis (Amidinopropane).
The pre-hemolytic mechanism induced by free radicals initiated from water-soluble 2,2'-azobis (2-amidinopropane) hydrochloride (AAPH) and its reversal by genistein was investigated in human erythrocytes. The time course of K+ efflux compared to the occurrence of hemolysis suggests that AAPH-induced hemolysis occurs indirectly via pore formation and band 3 oxidation as expected. However, genistein inhibited hemolysis, LDH release and membrane protein oxidation but not K+ efflux. This indicated that erythrocyte protein oxidation possibly in the hydrophobic core plays a significant role in the membrane pre-hemolytic damage. Chemiluminescence (CL) analysis carried out in non-lysed erythrocytes treated with AAPH showed a dramatic increase in CL indicating both reduced levels of antioxidants and increased membrane lipid peroxide. The V0 value was also increased up to 6 times, denoting a high degree of membrane peroxidation very early in erythrocyte membrane damage. The whole process was inhibited by genistein in a dose-dependent manner. These results indicate that the genistein inhibited both hemolysis and pre-hemolytic damage and also hindered membrane lipid peroxide formation and protein oxidation. In addition, it is suggested that pre-hemolytic damage is mediated mainly by the oxidation of both phospholipid and protein located in the deeper hydrophobic region of the membrane.
Colado Simão
Andréa Name
AN
Pathophysiology Laboratory of Free Radicals, Department of Pathological Sciences, Universidade Estadual de Londrina (UEL), 86051990 Londrina, Brazil.
Suzukawa
Andréia Akemi
AA
Casado
Maria Fernanda
MF
Oliveira
Rossinei Danieli
RD
Guarnier
Flávia Alessandra
FA
Cecchini
Rubens
R
D000578:Amidines / Q000633:toxicity*; D000975:Antioxidants / Q000494:pharmacology*; D004910:Erythrocyte Membrane / Q000187:drug effects / Q000378:metabolism; D004912:Erythrocytes / Q000187:drug effects* / Q000378:metabolism; D019833:Genistein / Q000494:pharmacology*; D005978:Glutathione / Q000378:metabolism; D006454:Hemoglobins / Q000378:metabolism; D006461:Hemolysis / Q000187:drug effects*; D006801:Humans; D066298:In Vitro Techniques; D008054:Lipid Peroxides / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects*; D011188:Potassium / Q000378:metabolism
D000578:Amidines; D000975:Antioxidants; D006454:Hemoglobins; D008054:Lipid Peroxides; C046728:2,2'-azobis(2-amidinopropane); D019833:Genistein; D005978:Glutathione; D011188:Potassium