Anti-platelet and anti-inflammatory properties of an ethanol-water red grape pomace extract

Abstract

<jats:title>Abstract</jats:title><jats:p>Previous reports support that wine contains a mixture of micro-constituents in a proper quality and quantity that possess cardio-protective effect partly through Platelet-Activating Factor (PAF) inhibition. Grape pomace (GP) is a source of wine-like micro-constituents that may be a suitable alternative in food fortification. Limited data exist concerning their effects on thrombosis and inflammation. Therefore, the purpose of this study was to examine a grape pomace extract regarding its anti-platelet and anti-inflammatory properties. GP from four red grape varieties were extracted with 80% ethanol (GP:solvent 1:5w/v). The extract's total phenolic compounds were evaluated and the phenolic profile was performed by Ultrahigh-Performance-Liquid-Chromatography coupled to Mass-Spectrometry and the determination of fatty acids profile was performed by Gas-Chromatography. The extract's anti-platelet properties were tested in healthy volunteers’ platelet rich plasma by the light transmittance method, against three agonists: PAF, ADP and TRAP. The results expressed as IC<jats:sub>50</jats:sub> values (μg of extract that cause 50% inhibition of aggregation) and EC<jats:sub>50</jats:sub> values (agonist concentration that causes 50% of the maximum aggregation) in the extract's presence and absence. Concerning the extract's anti-inflammatory properties, peripheral blood mononuclear cells from healthy volunteers were pre-incubated with different extract concentrations, which were tested for their effect on cell viability, for 1 h and then stimulated with LPS for 4 h. Secretion of IL-1β and TNF-a was measured and normalized with the total cell protein. Phenolic compounds were calculated at 8.79 ± 1.17 mg gallic acid per g of GP. The most abundant ones were catechin, epicatechin and quercetin at 202.9 ± 6.9, 84.8 ± 1.5 and 83.7 ± 3.5μg per g of GP respectively. Out of the 18 fatty acids detected, the most abundant ones were palmitic, oleic, linoleic, and linolenic acid at 28.7 ± 0.1, 11.4 ± 0.01, 32.5 ± 0.07, 12.7 ± 0.005 g per 100 g of fat. The extract's IC<jats:sub>50</jats:sub> was calculated at 162.1 ± 66.9, 181.2 ± 82.3 and 156.3 ± 97.5μg against PAF, ADP and TRAP, respectively. The EC<jats:sub>50</jats:sub> values in the presence of 150μg extract were increased (lower platelet aggregation sensitivity) approximately at 100%, 45% and 13% against PAF, ADP and TRAP respectively, compared to EC<jats:sub>50</jats:sub> values in the absence of extract. The presence of 500 and 1000μg/mL of extract reduced LPS-induced TNF-a secretion at approximately 38.2% (p = 0.04) and 6.0% (p &lt; 0.000), respectively. Potent anti-platelet and anti-inflammatory properties are combined in a grape pomace extract. The use of its bioactive micro-constituents is likely to lead to the production of functional foods with cardioprotective properties.</jats:p>

Laccase-Catalyzed Synthesis of Added-Value Polymers from Cork and Grape Extracts

“…The results reveal that a combination of ethanol and water extracts phenolic chemicals more efficiently than monocomponent solvent solutions. This can be related to the fact that adding a small amount of water to organic solvents generally results in a more polar media, as indicated by the polarity test, which promotes polyphenol extraction. , The addition of ethanol in combination with water may increase bioactive solubility because ethanol lowers the water surface tension, allowing for better penetration and mass transfer. Previous studies have shown that H 2 O/EtOH combinations (40–70%) remove phenolic chemicals more efficiently than water or ethanol. , From the data obtained, one can observe that 50:50 H 2 O/EtOH gave rise to higher extraction yields than water or ethanol, which agrees well with the previous work of Brand et al that reported high extraction yields of bioactives from cork with 100% ethanol, while 80% aqueous ethanol was the best solvent to extract grape .…”

“…This can be related to the fact that adding a small amount of water to organic solvents generally results in a more polar media, as indicated by the polarity test, which promotes polyphenol extraction. 38,39 The addition of ethanol in combination with water may increase bioactive solubility because ethanol lowers the water surface tension, allowing for better penetration and mass transfer. Previous studies have shown that H 2 O/EtOH combinations (40−70%) remove phenolic chemicals more efficiently than water or ethanol.…”

Winemaking: “With One Stone, Two Birds”? A Holistic Review of the Bio-Functional Compounds, Applications and Health Benefits of Wine and Wineries’ By-Products

“…Apart from the abundant phenolics, grapes, must, wine and winery by-products also contain lipid bioactive compounds, such as bioactive polar lipids and unsaturated fatty acids (UFA), with strong anti-inflammatory and antithrombotic properties, which contribute to the bio-functional activities and protective effects of grape products like wine and its by-products [1,[8][9][10][11][12][13][14][15][16][17][18][19][20][21][22][23]. In particular, some polar lipids (glycolipids and phospholipids) rich in UFA (i.e., oleic, linoleic and linolenic acids) are the most effective lipid bioactives against specific thrombo-inflammatory pathways, including those associated with PAF and thrombin [8][9][10][11].…”

“…In particular, some polar lipids (glycolipids and phospholipids) rich in UFA (i.e., oleic, linoleic and linolenic acids) are the most effective lipid bioactives against specific thrombo-inflammatory pathways, including those associated with PAF and thrombin [8][9][10][11]. Such bioactive lipid compounds pre-exist both in grapes and in yeasts, on which fermentation is based, and subsequently they are also found in both the remaining winery by-products [16,17,22,23] and in the final fermented wine product, while favourable modifications also occur during fermentation [8][9][10][11].…”

Attempts to Create Products with Increased Health-Promoting Potential Starting with Pinot Noir Pomace: Investigations on the Process and Its Methods

“…Additionally, given their bioactive properties, it has been suggested that polyphenolic compounds could be sourced as starting materials and converted into products that directly deliver health-promoting effects [ 4 , 19 ]. In the literature, there are a number of examples in which grape pomace extracts, which have higher or more concentrated phenolic contents, were found to exhibit antioxidant, antimicrobial, anti-inflammatory and antiplatelet activities [ 20 , 21 , 22 ], as well as promising antiproliferative or cytotoxic activities against cancer cells [ 23 , 24 ]. These studies were validated based on chemical, biochemical and in vitro studies and are promising indications for the health-promoting potential of the pomace.…”