Endothelial cells deconjugate resveratrol metabolites to free resveratrol: a possible role in tissue factor modulation

[Scope] The antithrombotic effects of resveratrol (RV) and its derivatives remain unknown. The objective is to evaluate the modulatory effects of RV, its glucoside form, piceid, and its biological metabolites (RV‐3‐O‐β‐d‐glucuronide, RV‐4’‐O‐d‐glucuronide, and RV‐3‐O‐sulfate) on tissue factor (TF). Moreover, the endothelial metabolism of RV is assessed. [Methods and results] Human aortic endothelial cells (HAECs) are incubated with trans‐piceid, trans‐RV, or their biological metabolites and stimulated with tumor necrosis factor‐α (TNF‐α). TF activity, protein levels, and mRNA expression are determined in cell lysates. Moreover, RV conjugation (phase‐II‐metabolism) to its sulfated or glucuronidated metabolites and their deconjugation to their parent compound (free RV) are also assessed in cell lysates and culture media. RV decreased TF activity, protein levels, and mRNA expression, whereas piceid and RV metabolites (RVmet) had no effects. RV‐3‐O‐sulfate was the main metabolite generated in the endothelium, while RVmet are deconjugated to free RV. Isomerization of trans‐RV and its trans‐metabolites to their cis‐forms is observed. [Conclusions] RV exerts antithrombotic effects by modulating TF. RVmet and piceid does not exert this effect. However, the capacity of endothelial cells to deconjugate RVmet to free RV indicates that RVmet function as an endothelial reservoir for RV regeneration, thus, contributing to the antithrombotic effects of RV.This work was supported by the CENIT program from the Spanish Ministry of Science and Innovation (Proyecto CENIT: MET‐DEV‐FUN; 2006–2009) and by the project AGL2016‐76943‐C2 from the Ministerio de Economía, Indústria y Competitividad, the Agencia Estatal de Investigación (AEI), and the European Regional Development Fund (ERDF). Ú.C. has a Pla Estratègic de Recerca i Innovació en Salut (PERIS) post‐doctoral grant (SLT002/16/00239; Catalonia, Spain) from Generalitat de Catalunya. S.F.‐C., Ú.C., and R.S. conceived and designed the experiment.Peer reviewe

The intake of olive oil phenolic compounds promotes macrophage-specific reverse cholesterol transport in vivo

Trabajo presentado en el 87th European Atherosclerosis Society (EAS) Congress, celebrado en Maastricht (Holanda), del 26 al 29 de mayo de 2019Background and Aims: In the present study, we determined the effects of the olive oil phenolic compounds on reverse cholesterol transport (RCT) from macrophages to feces in vivo. Methods: [3H]cholesterol-labeled J774 mouse macrophages were injected intraperitoneally into C57BL/6 mice given intragastric doses of refined olive oil (ROO), virgin olive oil enriched with their own phenolic compounds (FVOO, 500ppm), the phenolic compounds (PCs, 500 ppm) resuspended in saline or the saline solution for 14 days and radioactivity was determined in plasma, liver, and feces collected for 48 hours. The amount of preß-HDL was quantified with two-dimensional crossed immunoelectrophoresis. The cholesterol efflux capacity of 3% apoBdepleted plasma samples was determined by using J774 mouse macrophages labeled with TopFluor-cholesterol. RT-PCR assays were performed on a CFX96TM Real-Time System. Results: FVOO caused a significant increase in HDL cholesterol, apoA-I, HDL size and the formation of nascent preb-HDL particles. These changes were concomitant with enhanced macrophage-derived [3H] cholesterol eflux to feces of mice in vivo. PCs intake also promoted macrophage-to-feces RCT compared with that of ROO and vehicle groups. Unlike ROO or vehicle intake, FVOO and PCs intake increased ex vivo macrophage cholesterol efflux, which was closely associated with HDL cholesterol levels. However, the expression of the main liver genes involved in RCT was not affected by FVOO and PCs intake. Conclusions: Consumption of a functional olive oil, enriched with its own PCs, enhances macrophage-specific RCT in vivo. Our data provide direct in vivo evidence of the crucial role of PCs in the induction of macrophagespecific RCT

Impact of Virgin Olive Oil and Phenol-Enriched Virgin Olive Oils on the HDL Proteome in Hypercholesterolemic Subjects: A Double Blind, Randomized, Controlled, Cross-Over Clinical Trial (VOHF Study)

The effects of olive oil phenolic compounds (PCs) on HDL proteome, with respect to new aspects of cardioprotective properties, are still unknown. The aim of this study was to assess the impact on the HDL protein cargo of the intake of virgin olive oil (VOO) and two functional VOOs, enriched with their own PCs (FVOO) or complemented with thyme PCs (FVOOT), in hypercholesterolemic subjects. Eligible volunteers were recruited from the IMIM-Hospital del Mar Medical Research Institute (Spain) from April 2012 to September 2012. Thirty-three hypercholesterolemic participants (total cholesterol >200mg/dL; 19 men and 14 women; aged 35 to 80 years) were randomized in the double-blind, controlled, cross-over VOHF clinical trial. The subjects received for 3 weeks 25 mL/day of: VOO, FVOO, or FVOOT. Using a quantitative proteomics approach, 127 HDL-associated proteins were identified. Among these, 15 were commonly differently expressed after the three VOO interventions compared to baseline, with specific changes observed for each intervention. The 15 common proteins were mainly involved in the following pathways: LXR/RXR activation, acute phase response, and atherosclerosis. The three VOOs were well tolerated by all participants. Consumption of VOO, or phenol-enriched VOOs, has an impact on the HDL proteome in a cardioprotective mode by up-regulating proteins related to cholesterol homeostasis, protection against oxidation and blood coagulation while down-regulating proteins implicated in acute-phase response, lipid transport, and immune response. The common observed protein expression modifications after the three VOOs indicate a major matrix effect

Effect of virgin olive oil and thyme phenolic compounds on blood lipid profile: implications of human gut microbiota.

PURPOSE: To investigate the effect of virgin olive oil phenolic compounds (PC) alone or in combination with thyme PC on blood lipid profile from hypercholesterolemic humans, and whether the changes generated are related with changes in gut microbiota populations and activities. METHODS: A randomized, controlled, double-blind, crossover human trial (n = 12) was carried out. Participants ingested 25 mL/day for 3 weeks, preceded by 2-week washout periods, three raw virgin olive oils differing in the concentration and origin of PC: (1) a virgin olive oil (OO) naturally containing 80 mg PC/kg, (VOO), (2) a PC-enriched virgin olive oil containing 500 mg PC/kg, from OO (FVOO), and (3) a PC-enriched virgin olive oil containing a mixture of 500 mg PC/kg from OO and thyme, 1:1 (FVOOT). Blood lipid values and faecal quantitative changes in microbial populations, short chain fatty acids, cholesterol microbial metabolites, bile acids, and phenolic metabolites were analysed. RESULTS: FVOOT decreased seric ox-LDL concentrations compared with pre-FVOOT, and increased numbers of bifidobacteria and the levels of the phenolic metabolite protocatechuic acid compared to VOO (P < 0.05). FVOO did not lead to changes in blood lipid profile nor quantitative changes in the microbial populations analysed, but increased the coprostanone compared to FVOOT (P < 0.05), and the levels of the faecal hydroxytyrosol and dihydroxyphenylacetic acids, compared with pre-intervention values and to VOO, respectively (P < 0.05). CONCLUSION: The ingestion of a PC-enriched virgin olive oil, containing a mixture of olive oil and thyme PC for 3 weeks, decreases blood ox-LDL in hypercholesterolemic humans. This cardio-protective effect could be mediated by the increases in populations of bifidobacteria together with increases in PC microbial metabolites with antioxidant activities.This work was supported by Instituto de Salud Carlos III FEDER (RD12-0042, CB06/03/0028, CD10/00224, CP06/00100, CB06/02/0029, CA11/00215), Ministry of Economy and Competitiveness (AGL2012-40144-C03-01, AGL2012-40144-C03-02, AGL2012-40144-C03-03, FPI:BES- 2010-040766), Agency for Management of University and Research Grants (2009 SGR 1195, 2014 SGR 240). We thank M Angels Calvo for the growth of pure cultures, Malén Massot for helping us in the elaboration of the FISH-FC protocol, Óscar Fornas and Cristina Llop for their technical assistance and Borges Mediterranean Group.http://dx.doi.org/10.1007/s00394-015-1063-

Effect of virgin olive oil and thyme phenolic compounds on blood lipid profile: implications of human gut microbiota.

PURPOSE: To investigate the effect of virgin olive oil phenolic compounds (PC) alone or in combination with thyme PC on blood lipid profile from hypercholesterolemic humans, and whether the changes generated are related with changes in gut microbiota populations and activities. METHODS: A randomized, controlled, double-blind, crossover human trial (n = 12) was carried out. Participants ingested 25 mL/day for 3 weeks, preceded by 2-week washout periods, three raw virgin olive oils differing in the concentration and origin of PC: (1) a virgin olive oil (OO) naturally containing 80 mg PC/kg, (VOO), (2) a PC-enriched virgin olive oil containing 500 mg PC/kg, from OO (FVOO), and (3) a PC-enriched virgin olive oil containing a mixture of 500 mg PC/kg from OO and thyme, 1:1 (FVOOT). Blood lipid values and faecal quantitative changes in microbial populations, short chain fatty acids, cholesterol microbial metabolites, bile acids, and phenolic metabolites were analysed. RESULTS: FVOOT decreased seric ox-LDL concentrations compared with pre-FVOOT, and increased numbers of bifidobacteria and the levels of the phenolic metabolite protocatechuic acid compared to VOO (P < 0.05). FVOO did not lead to changes in blood lipid profile nor quantitative changes in the microbial populations analysed, but increased the coprostanone compared to FVOOT (P < 0.05), and the levels of the faecal hydroxytyrosol and dihydroxyphenylacetic acids, compared with pre-intervention values and to VOO, respectively (P < 0.05). CONCLUSION: The ingestion of a PC-enriched virgin olive oil, containing a mixture of olive oil and thyme PC for 3 weeks, decreases blood ox-LDL in hypercholesterolemic humans. This cardio-protective effect could be mediated by the increases in populations of bifidobacteria together with increases in PC microbial metabolites with antioxidant activities.This work was supported by Instituto de Salud Carlos III FEDER (RD12-0042, CB06/03/0028, CD10/00224, CP06/00100, CB06/02/0029, CA11/00215), Ministry of Economy and Competitiveness (AGL2012-40144-C03-01, AGL2012-40144-C03-02, AGL2012-40144-C03-03, FPI:BES- 2010-040766), Agency for Management of University and Research Grants (2009 SGR 1195, 2014 SGR 240). We thank M Angels Calvo for the growth of pure cultures, Malén Massot for helping us in the elaboration of the FISH-FC protocol, Óscar Fornas and Cristina Llop for their technical assistance and Borges Mediterranean Group.http://dx.doi.org/10.1007/s00394-015-1063-