Oxidative damage of vascular smooth muscle cells by the glycated protein-cupric ion system

Atherosclerosis
N SakataT Furukawa

Abstract

To clarify the mechanism of cellular injury through the nonenzymatic reaction of glucose with proteins, we studied the cytotoxic effect of glycated bovine serum albumin on cultured smooth muscle cells in the presence of cupric ion. Glycated proteins were prepared by incubating bovine serum albumin with 0.5 M D-glucose in 0.3 M sodium phosphate buffer at 37 degrees C for 2, 4 and 16 weeks (g-BSA-2, g-BSA-4 and g-BSA-16, respectively). Early glycation products, such as fructosamine, were formed more than two weeks after incubation. However, the immunoreactivity of glycated proteins to anti-AGE antibody was 12-fold higher in g-BSA-16 than in g-BSA-2. Both g-BSA-2 and g-BSA-16 showed a concentration-dependent cytotoxicity in smooth muscle cells in the presence of 80 microM cupric ion by an MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) dye reduction assay and dye exclusion test. Flow cytometry and spectrofluorophotometry using dihydrorhodamine 123 showed that the extracellular generation of oxidants was dose-dependently enhanced with increasing concentrations of g-BSA-2 or g-BSA-16 in the presence of cupric ion. However, no difference was observed in the intracellular generation of oxidants between the presence...Continue Reading

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Citations

Oct 17, 2001·Clinical Chemistry and Laboratory Medicine : CCLM·V CosićV B Djordjević
Nov 26, 2008·Journal of Diabetes and Its Complications·Maryam Bassirat, Zeinab Khalil
Jan 2, 2008·Toxicology in Vitro : an International Journal Published in Association with BIBRA·Radamés Alemón-MedinaIsabel Gracia-Mora
Apr 24, 2016·Annals of Biomedical Engineering·Laura Hockaday KangJonathan Talbot Butcher
Aug 2, 2001·Journal of Atherosclerosis and Thrombosis·N SakataS Takebayashi

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