Combined Treatment of Hydroxytyrosol with Carbon Monoxide-Releasing Molecule-2 Prevents TNFα-Induced Vascular Endothelial Cell Dysfunction through NO Production with Subsequent NFκB Inactivation
Figure 5
Combined effects of HT and CORM-2 on eNOS phosphorylation and NO production in VECs. (a) Serum-starved VECs were incubated with 50 μM HT or 50 μM CORM-2 for the indicated periods (0 to 12 h). Phospho-eNOS (Ser1177) protein levels were determined using western blotting and normalized to the amount of α-tubulin. Representative data is shown from three independent experiments that yielded similar results. * versus untreated cells. (b) Serum-starved VECs were treated with 50 μM HT in the presence of 50 μM CORM-2 for 1 h. Phospho-eNOS (Ser1177) protein levels were determined using western blotting and normalized to the amount of α-tubulin. Representative data is shown from three independent experiments that yielded similar results. * versus untreated cells. (c) Serum-starved VECs were treated with 50 μM HT or 50 μM CORM-2 in the presence or absence of 1 mM L-NMMA for 3, 6, and 24 h. NO production was assessed using the fluorescent reagent DAF-2 DA and expressed as a percentage of the control. Data represent the mean ± SD of three different independent experiments (*).
