Protection of erythrocytes against oxidative damage and autologous immunoglobulin G (IgG) binding by iron chelator fluor-benzoil-pyridoxal hydrazone.
Iron is released in a free desferrioxamine-chelatable form when erythrocytes are challenged by an oxidative stress. The release of iron is believed to play an important role in inducing destructive damage (lipid peroxidation and hemolysis) or in producing membrane protein oxidation and generation of senescent cell antigens (SCA). In this report, we further tested the hypothesis that intracellular chelation of iron released under conditions of oxidative stress prevents erythrocyte damage or SCA formation. Fluor-benzoil-pyridoxal hydrazone (FBPH), an iron-chelating molecule of the family of aromatic hydrazones, was prepared by synthesis and used for the above purpose after the capacity of the product to enter cells had been ascertained. GSH-depleted mouse erythrocytes were incubated with the oxidant drug phenylhydrazine in order to produce iron release, lipid peroxidation, and hemolysis. FBPH at a concentration of 200 microM prevented lipid peroxidation and hemolysis in spite of equal values of iron release. FBPH was active even at a lower concentration (100 microM) when the erythrocytes were preincubated with it for 15 min. No preventive effect was seen when FBPH saturated with iron was used. Prolonged aerobic incubation (60 hr) of erythrocytes produced iron release and formation of SCA as determined by autologous immunoglobulin G (IgG) binding. The IgG binding was detected by using an anti-IgG antibody labeled with fluorescein and by examining the cells for fluorescence by confocal microscopy. FBPH prevented SCA formation in a dose-related manner. These results lend further support to the hypothesis that iron release is a key factor in erythrocyte ageing.
Ferrali
M
M
Department of Pathophysiology and Experimental Medicine, University of Siena, 53100, Siena, Italy.
Signorini
C
C
Ciccoli
L
L
Bambagioni
S
S
Rossi
V
V
Pompella
A
A
Comporti
M
M
D000818:Animals; D000943:Antigens, Differentiation / Q000276:immunology; D004912:Erythrocytes / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D006835:Hydrazones / Q000494:pharmacology*; D007074:Immunoglobulin G / Q000276:immunology / Q000378:metabolism*; D066298:In Vitro Techniques; D007501:Iron / Q000378:metabolism*; D007502:Iron Chelating Agents / Q000494:pharmacology*; D051379:Mice; D018384:Oxidative Stress; D020011:Protective Agents / Q000494:pharmacology; D011730:Pyridoxal / Q000031:analogs & derivatives* / Q000494:pharmacology
D000943:Antigens, Differentiation; D006835:Hydrazones; D007074:Immunoglobulin G; D007502:Iron Chelating Agents; D020011:Protective Agents; C047076:senescent cell differentiation antigen; C065798:pyridoxal 3-fluorobenzoyl hydrazone; D011730:Pyridoxal; D007501:Iron