WINE POMACE EXTRACT PREVENTS INFLAMMATORY RESPONSE IN ENTEROCYTE-LIKE CELLS AND IMPROVES VASCULAR ENDOTHELIAL FUNCTIONS

Inflammatory bowel diseases (IBD) are chronic inflammatory conditions involving primarily the gastrointestinal tract, but also associated with systemic vascular manifestations. In IBD patients, the incidence of cardiovascular disease appears to be increased due to chronic inflammation, which affects arterial properties and causes endothelial dysfunction. Polyphenols are anti-oxidant phytochemical compounds able to counteract inflammation and endothelial dysfunction. Winemaking waste contains natural bioactive compounds, including polyphenols, therefore the use of pomace extracts could be a low-cost eco-sustainable strategy to prevent intestinal and vascular diseases. The aim of this study was to determine whether wine pomace polyphenolic extract (WPE) was able to mitigate inflammatory response in enterocyte-like cells and to improve vascular functions. To evaluate the anti-inflammatory effects of WPE, a co-culture model of intestinal epithelial/endothelial cells (Caco-2/HMEC-1) was used. Caco-2 cells were plated on the upper side of transwell inserts and HMEC-1 in the basolateral compartment. Caco-2 cells were treated with different concentrations of WPE (1, 5, 10 ?g GAEs/mL) for 2h and then stimulated with lipopolysaccharide (LPS) and tumour necrosis factor(TNF)-? for 16h. Through multiple assays, the expression of intestinal and endothelial inflammatory mediators, intracellular ROS levels, NF-?B activity as well as the endothelial-monocyte adhesion were analyzed. WPE supplementation significantly prevented, in a concentration dependent manner, interleukin(IL)-6 and monocyte chemoattractant protein(MCP)-1 production, in Caco-2 cells. Moreover, WPE suppressed the gene expression of cytokines (IL-6, IL-1?, TNF-?), chemokines (MCP-1, M-CSF, CXCL-10) as well as adhesion molecules (ICAM-1, VCAM-1) and matrix metalloproteinases (MMP)-2 and -9. The decreased inflammatory gene expression was related to the inhibition of NF-?B activity and reduced intracellular ROS levels. Furthermore, in co-culture Caco-2/HMEC-1, WPE reduced endothelial cell activation and monocyte adhesion to inflamed HMEC-1, with lowered levels of adhesion molecules (ICAM-1, VCAM-1), cytokines (IL-6, IL-1?), and chemokines (MCP-1, CXCL-10). These findings demonstrated that WPE, as a natural source of polyphenols with multiple healthy properties, could contribute to mitigate chronic gut inflammatory diseases and improve vascular endothelial functions.