Antioxidant and anti-inflammatory effects of Rosmarinus officinalis and Melissa officinalis ethanolic extracts

Abstract

Background and Aims : Oxidized LDL, endothelial dysfunction and neutrophil extracellular trap formation (NETs) play a central role in atherosclerosis. The aim of the present study was to investigate the effect of Rosmarinus officinalis (ROe) and Melissa officinalis (MOe) ethanolic extracts on LDL oxidation, intercellular adhesion molecule-1 (ICAM-1) expression on endothelial cells (HUVECs) and NET formation.Methods: Ethanolic extraction of whole plants was performed for 72h. Phytochemical analysis was performed using NMR-spectroscopy. LDL (d=1.019-1.063g/mL) was isolated by sequential ultracentrifugation of plasma from healthy volunteers and oxidized (100μg protein/mL) by 5μM CuSO4, in the presence of ROe or MOe, for 6h. HUVECs were pre-incubated with various extract concentrations for 1h and treated with 0.5ng/mL TNF-α for 6h or 8U/mL thrombin for 24h. ICAM-1 expression was determined by flow cytometry. Neutrophils (2×105) were isolated, pre-incubated with the extracts for 5min and activated with 100nM PMA for 4h at 37°C, 5% CO2. NETs were isolated and quantitated using ELISA kit.Conclusions: ROe and MOe share many similarities in their phytochemical profile, however ROe exhibits more potent antioxidant activity than MOe and anti-inflammatory activities that are not present in MOe. Further ongoing studies will shed light into the active compounds of ROe and their specific mechanisms of action. Background and Aims : Oxidized LDL, endothelial dysfunction and neutrophil extracellular trap formation (NETs) play a central role in atherosclerosis. The aim of the present study was to investigate the effect of Rosmarinus officinalis (ROe) and Melissa officinalis (MOe) ethanolic extracts on LDL oxidation, intercellular adhesion molecule-1 (ICAM-1) expression on endothelial cells (HUVECs) and NET formation. Methods: Ethanolic extraction of whole plants was performed for 72h. Phytochemical analysis was performed using NMR-spectroscopy. LDL (d=1.019-1.063g/mL) was isolated by sequential ultracentrifugation of plasma from healthy volunteers and oxidized (100μg protein/mL) by 5μM CuSO4, in the presence of ROe or MOe, for 6h. HUVECs were pre-incubated with various extract concentrations for 1h and treated with 0.5ng/mL TNF-α for 6h or 8U/mL thrombin for 24h. ICAM-1 expression was determined by flow cytometry. Neutrophils (2×105) were isolated, pre-incubated with the extracts for 5min and activated with 100nM PMA for 4h at 37°C, 5% CO2. NETs were isolated and quantitated using ELISA kit. Conclusions: ROe and MOe share many similarities in their phytochemical profile, however ROe exhibits more potent antioxidant activity than MOe and anti-inflammatory activities that are not present in MOe. Further ongoing studies will shed light into the active compounds of ROe and their specific mechanisms of action.