BACKGROUND: Inflammation is a common process in endothelial dysfunction, characterized by an increase in the expression of endothelial cell adhesion molecules and a decrease of nitric oxide production [1]. Evidence suggests that polyphenols may play a beneficial role in attenuating inflammation with implications on atherosclerosis [2]. However, their mechanism of action is still not entirely understood due to their poor absorption and extensive intestinal and hepatic transformation [3]. OBJECTIVE: This study aims to evaluate the ability of some gut phenolic metabolites (i.e. protocatechuic, gallic, syringic and vanillic acid; PA, GA, SA, VA, respectively) to reduce the adhesion of monocytes (THP-1) to endothelial cells (HUVECs), in a stimulated pro-inflammatory environment, and to decrease the production of cell adhesion molecules (VCAM-1 and E-selectin), as potential markers involved in such modulation. METHODS: Adhesion of THP-1 to HUVECs: Day 1-Preparation of 96 wells plate (2x104 HUVEC per well); Day 2-Labelling of THP-1 cells with CellTrackerTM Green CMFDA, addition of THP-1 (2x105 cells/well) and TNF- (100 ng mL-1) to HUVEC, and incubation for 24h; Day 3-Incubation with PA, GA, VA and SA at different concentrations (from 0.01 till 10μg mL-1) for 24 h; Day 4- Reading of the fluorescence (excitation: 485 nm, emission: 538 nm, mod. F200 Infinite, TECAN Milan, Italy) Evaluation of cell adhesion molecules: The levels of vascular cell adhesion molecules (VCAM-1), and endothelial selectin (E-selectin) were measured in the supernatant by ELISA kits following the manufacture’s instruction. The results derived from three indipendent experiments. RESULTS: GA and PA significantly decreased the adhesion of THP-1 to HUVECs at 1 µg mL-1 (-18.0%; p=0.04 for GA, -29.5%; p=0.03 for PA) and 10 µg mL-1 (-59.3%; p<0.001 for GA, -44.3%; p=0.001 for PrA) compared to TNF-. VA significantly reduced the adhesion only at the maximum concentration (-20.8%; p<0.005). No effect was observed after SA supplementation. CONCLUSION: In conclusion, these preliminary results seem to support the capacity of gut phenolic metabolites to modulate the endothelial cell adhesion process. The reduction in the adhesion process seems to be driven by a decrease of E-selectin levels but not VCAM-1. Ongoing experiments are attempting to clarify the mechanisms of action of each compound involved in the above observations.

In vitro approach to evaluate the role of gut phenolic metabolites in the modulation of inflammation and atherosclerosis / C. DEL BO', M. Marino, P. Riso, K. Dorothy, M. Porrini. ((Intervento presentato al 5. convegno Foodomics Conference "Foodomics 2018 : From Data to Knowledge" tenutosi a Cesena nel 2018.

In vitro approach to evaluate the role of gut phenolic metabolites in the modulation of inflammation and atherosclerosis

Cristian Del Bo’¹;Mirko Marino¹;Patrizia Riso¹;Marisa Porrini¹
2018-01-11

Abstract

BACKGROUND: Inflammation is a common process in endothelial dysfunction, characterized by an increase in the expression of endothelial cell adhesion molecules and a decrease of nitric oxide production [1]. Evidence suggests that polyphenols may play a beneficial role in attenuating inflammation with implications on atherosclerosis [2]. However, their mechanism of action is still not entirely understood due to their poor absorption and extensive intestinal and hepatic transformation [3]. OBJECTIVE: This study aims to evaluate the ability of some gut phenolic metabolites (i.e. protocatechuic, gallic, syringic and vanillic acid; PA, GA, SA, VA, respectively) to reduce the adhesion of monocytes (THP-1) to endothelial cells (HUVECs), in a stimulated pro-inflammatory environment, and to decrease the production of cell adhesion molecules (VCAM-1 and E-selectin), as potential markers involved in such modulation. METHODS: Adhesion of THP-1 to HUVECs: Day 1-Preparation of 96 wells plate (2x104 HUVEC per well); Day 2-Labelling of THP-1 cells with CellTrackerTM Green CMFDA, addition of THP-1 (2x105 cells/well) and TNF- (100 ng mL-1) to HUVEC, and incubation for 24h; Day 3-Incubation with PA, GA, VA and SA at different concentrations (from 0.01 till 10μg mL-1) for 24 h; Day 4- Reading of the fluorescence (excitation: 485 nm, emission: 538 nm, mod. F200 Infinite, TECAN Milan, Italy) Evaluation of cell adhesion molecules: The levels of vascular cell adhesion molecules (VCAM-1), and endothelial selectin (E-selectin) were measured in the supernatant by ELISA kits following the manufacture’s instruction. The results derived from three indipendent experiments. RESULTS: GA and PA significantly decreased the adhesion of THP-1 to HUVECs at 1 µg mL-1 (-18.0%; p=0.04 for GA, -29.5%; p=0.03 for PA) and 10 µg mL-1 (-59.3%; p<0.001 for GA, -44.3%; p=0.001 for PrA) compared to TNF-. VA significantly reduced the adhesion only at the maximum concentration (-20.8%; p<0.005). No effect was observed after SA supplementation. CONCLUSION: In conclusion, these preliminary results seem to support the capacity of gut phenolic metabolites to modulate the endothelial cell adhesion process. The reduction in the adhesion process seems to be driven by a decrease of E-selectin levels but not VCAM-1. Ongoing experiments are attempting to clarify the mechanisms of action of each compound involved in the above observations.
Settore MED/49 - Scienze Tecniche Dietetiche Applicate
In vitro approach to evaluate the role of gut phenolic metabolites in the modulation of inflammation and atherosclerosis / C. DEL BO', M. Marino, P. Riso, K. Dorothy, M. Porrini. ((Intervento presentato al 5. convegno Foodomics Conference "Foodomics 2018 : From Data to Knowledge" tenutosi a Cesena nel 2018.
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/2434/593252
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