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 4
Combined effect of HT and CORM-2 on TNFα-mediated NFκB activation. (a) Serum-starved VECs were treated with 50 μM iCORM-2 or 50 μM CORM-2 for 1 h in the presence of HT prior to the addition of TNFα (2 ng/mL) for 3 h. Phospho-NFκBp65 (Ser536) and phospho-IκBα (Ser32) 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. (b) Serum-starved VECs were treated with HT and CORM-2 (50 μM) for 1 h followed by TNFα (2 ng/mL) stimulation for 3 h. NFκBp65 expression was determined using western blotting in both cytosolic and nuclear fractions. Cytosolic and nuclear fractions were blotted for α-tubulin and Lamin B, respectively, for the protein normalization. Representative data is shown from three independent experiments that yielded similar results.
