AGE-BSA increased NADPH-dependent intracellular ROS production in HUCLs.

<p>Intracellular ROS production was assessed using a DCFH-DA fluorescence. (A) HUCLs were treated with AGE-BSA (200 µg/ml) for 3, 6, 12 and 24 h, Hydrogen peroxide (1 mM) was added to cells as a positive control. (B) Intracellular ROS generation was visualized under a laser scanning confocal microscope. Scale bar = 50 µm. (C) HUCLs were pretreated with DPI (NADPH oxidase inhibitor, 10 µM), apocynin (NADPH oxidase inhibitor, 300 µM), allopurinol (xanthine oxidase inhibitor, 10 µM), rotenone (inhibitor of mitochondrial electron transport complex I, 5 µM) or neutralizing anti-RAGE antibodies (20 µg/ml) for 1 h. and then incubated with AGE-BSA (200 µg/ml) for 12 h. Data are representative of three independent experiments. *, <i>P</i><0.05 vs. medium alone group; #, <i>P</i><0.05 vs. AGE-BSA group.