Hydroxytyrosol reduces intracellular reactive oxygen species levels in vascular endothelial cells by upregulating catalase expression through the AMPK-FOXO3a pathway.

Reactive oxygen species are critically involved in the endothelial dysfunction that contributes to atherosclerosis development. Hydroxytyrosol (HT), a main phenolic compound in olive oil and leaves from Olea europaea L., has antiatherogenic properties with powerful antioxidant activity. The present study verifies the antioxidant activity of HT on H2O2-induced intracellular reactive oxygen species in porcine pulmonary artery endothelial cells (VECs) and the involved molecular mechanisms. Incubation of VECs with HT prevented the increase in intracellular reactive oxygen species levels in the presence of H2O2. HT increased catalase mRNA, protein and activity. Catalase siRNA suppressed HT-dependent reduction of intracellular reactive oxygen species. HT increased both cytosolic and nuclear protein levels of forkhead transcription factor 3a (FOXO3a), as well as the phosphorylation of AMP-activated protein kinase (AMPK) that translocates FOXO3a to the nucleus. AMPK siRNA and a specific inhibitor suppressed HT-induced FOXO3a upregulation and catalase expression. Moreover, FOXO3a siRNA blocked HT-dependent increase in catalase expression. Taken together, our findings strongly demonstrate that HT positively regulates the antioxidant defense system in VECs by inducing the phosphorylation of AMPK with subsequent activation of FOXO3a and catalase expression, and provides a molecular basis for the prevention of cardiovascular diseases by HT.

Hydroxytyrosol induces proliferation and cytoprotection against oxidative injury in vascular endothelial cells: role of Nrf2 activation and HO-1 induction.

Hydroxytyrosol (HT), a phenolic compound in olive oil and leaves, has been reported to prevent various human pathologies including cardiovascular diseases. This study investigated the effects of HT on proliferation and protection against oxidative stress-induced damage in vascular endothelial cells (VECs) and the molecular mechanism(s) involved. Treatment of VECs with HT increased cell proliferation, promoted wound repair, and protected cells against H(2)O(2) cytotoxicity through the activation of Akt and ERK1/2, but not p38 MAPK. HT increased the expression and nuclear translocation of nuclear factor-E2-related factor-2 (Nrf2). Nrf2 expression was attenuated by LY294002 and U0126, inhibitors of phosphatidylinositol-3-kinase and MEK1/2, respectively. Nrf2 siRNA decreased both proliferative and cytoprotective effects of HT and abrogated HO-1 induction. Moreover, HO-1 inhibition with HO-1 siRNA or zinc protoporphyrin IX significantly prevented HT-induced cell proliferation, cytoprotection, and reduction in intracellular reactive oxygen species (ROS), suggesting that HO-1 is involved in these HT functions. The findings demonstrate that HT positively regulates the antioxidant defense system in VECs through the activation of Nrf2 followed by cell proliferation and resistance to vascular injury. The present study provides a molecular basis for the contribution of HT in the Mediterranean diet to the prevention of cardiovascular diseases.