title: Genistein abrogates pre-hemolytic and oxidative stress damage induced by 2,2'-Azobis (Amidinopropane).
abstract: 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.
authors.affiliation: Pathophysiology Laboratory of Free Radicals, Department of Pathological Sciences, Universidade Estadual de Londrina (UEL), 86051990 Londrina, Brazil.
authors.forename: Andréa Name
authors.initials: AN
authors.lastname: Colado Simão
authors.forename: Andréia Akemi
authors.initials: AA
authors.lastname: Suzukawa
authors.forename: Maria Fernanda
authors.initials: MF
authors.lastname: Casado
authors.forename: Rossinei Danieli
authors.initials: RD
authors.lastname: Oliveira
authors.forename: Flávia Alessandra
authors.initials: FA
authors.lastname: Guarnier
authors.forename: Rubens
authors.initials: R
authors.lastname: Cecchini
mesh_terms: 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
chemical_list: D000578:Amidines; D000975:Antioxidants; D006454:Hemoglobins; D008054:Lipid Peroxides; C046728:2,2'-azobis(2-amidinopropane); D019833:Genistein; D005978:Glutathione; D011188:Potassium