Flavonoids and platelet aggregation: A brief review

European Journal of Pharmacology 807 (2017) 91–101

Contents lists available at ScienceDirect

European Journal of Pharmacology journal homepage: www.elsevier.com/locate/ejphar

Flavonoids and platelet aggregation: A brief review a,⁎

b,c

a

MARK d,e

f,g

Caterina Faggio , Antoni Sureda , Silvia Morabito , Ana Sanches-Silva , Andrei Mocan , Seyed Fazel Nabavih, Seyed Mohammad Nabavih,⁎⁎ a Department of Biological, Pharmaceutical and Environmental Sciences, University of Messina, Viale Ferdinando Stagno d'Alcontres, 31 98166 S.Agata, Messina, Italy b Research Group on Community Nutrition and Oxidative Stress (NUCOX), University of Balearic Islands, E-07122 Palma de Mallorca, Balearic Islands, Spain c CIBEROBN (Physiopathology of Obesity and Nutrition), E-07122 Palma de Mallorca, Balearic Islands, Spain d National Institute of Health Dr. Ricardo Jorge, I.P., Department of Food and Nutrition, Av. Padre Cruz, 1649-016 Lisbon, Portugal e Centro de Estudos de Ciência Animal (CECA), ICETA – Instituto de Ciências, Tecnologias e Agroambiente da Universidade do Porto, Universidade do Porto – Praça Gomes Teixeira, Apartado 55142, 4051-401 Oporto, Portugal f Department of Pharmaceutical Botany, Faculty of Pharmacy, University of Medicine and Pharmacy “Iuliu Hațieganu”, Ghe. Marinescu 23, 400337 ClujNapoca, Romania g ICHAT and Institute for Life Sciences, University of Agricultural Sciences and Veterinary Medicine, Calea Mănăs ̧tur 3-5, 400372 Cluj-Napoca, Romania h Applied Biotechnology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran

A R T I C L E I N F O

A BS T RAC T

Keywords: Flavonoids Phytochemical Platelet aggregation Polyphenols

Platelets are small anucleated fragments derived from a megakaryocyte precursor. Platelets play a key role in many physiological functions especially in hemostasis and wound healing processes in order to maintain the integrity of the circulatory system. In addition, activated platelets release cytokines and chemokines which modulate the immune response and, in some cases of hyperactivation, they could be associated to the pathogenesis of inflammatory diseases. Flavonoids are polyphenolic compounds ubiquosly found in plants known to be potent antioxidants with positive effects against diverse diseases such as cancer, neurodegenerative or cardiovascular disease. It has been reported that some flavonoids possess anti-platelet aggregation effects though different pathways, being the inhibition of the arachidonic acid-based pathway the most representative mechanism of action. In the present review, the main sources of flavonoids, as well as their bioavailability and metabolism are summarized. Moreover, the available data about the anti-aggregation effects of flavonoids and the different mechanisms of action that has been proposed until now are also discussed.

1. Introduction Platelets are irregular, anucleated disc-shaped elements produced from large bone marrow cells called megakaryocytes, having a characteristic diameter of ~2–3 µm. Approximately, 1×1011 platelets per day are generated under normal conditions with a mean half-life between 7 and 10 days in circulation. (George, 2000). The membrane of platelets contains glycoproteins with a central role in the modulation of adhesive and cohesive platelet functions via interaction with different ligands (White and Clawson, 1980; White et al., 1999; Ebbeling et al., 1992; Rendu and Brohard-Bohn, 2001). Platelets include three main types of granules in the cytoplasm, including α, dense, and lysosomal granules. Upon activation these granules act as secretory vesicles, releasing their content to the

extracellular fluid. Plateletet granules contain a wide range of proteins including coagulation factors, adhesive proteins, chemokines, growth factors, proteoglycans, proteases and protease inhibitors with a paracrine and autocrine activity (Thon et al., 2012). α-Granules are the most numerous platelet granules and also contain most of the factors implicated in hemostasis, thrombosis and adhesion molecules which are involved in the interactions between platelets and the vessel wall (Maynard et al., 2007). Moreover, αgranules also include proteins related to processes like the inflammatory process, wound healing, mitogenic growth factors and a broad range of chemokines (Zufferey et al., 2014). Dense granules are smaller and less numerous granules, which release constituents contributing to engage additional platelets (in the aggregation process) and to local vasoconstriction by releasing several mediators (e.g., serotonin)

⁎ Correspondence to: Department of Chemical, Biological, Pharmaceutical and Environmental Sciences, University of Messina, Viale Ferdinando Stagno d′Alcontres, 31-98166 S.Agata, Messina, Italy. ⁎⁎ Corresponding author. E-mail addresses: [email protected] (C. Faggio), [email protected] (S.M. Nabavi).

http://dx.doi.org/10.1016/j.ejphar.2017.04.009 Received 19 October 2016; Received in revised form 4 April 2017; Accepted 10 April 2017 Available online 13 April 2017 0014-2999/ © 2017 Elsevier B.V. All rights reserved.

European Journal of Pharmacology 807 (2017) 91–101

C. Faggio et al.

pathway produces chalcones, and from these various flavonoids are formed. Depending on the plant, different groups of enzymes (e.g. reductases, hydroxylases, and isomerases) can act and originate different flavonoids (Ferreyra et al., 2012). Flavonoids can be divided in 2-phenylchromans (flavonols, flavones, flavanones, flavan-3-ols, anthocyanidins and condensed tannins) and 3-phenylchromans (isoflavonoids). Chalcones, although having a similar structure (they aromatic ketones) they are not true flavonoids. Flavonoids are known to be potent antioxidants and free radical scavengers derived from a double bond situated between carbons two and three, a hydroxyl group in carbon position three, poly-hydroxylation of the aromatic rings A and B aromatic rings and a carbonyl group located in the carbon four (Cook and Samman, 1996; Nabavi et al., 2012, 2015). Table 1 shows the physico-chemicals properties of the main representatives of some classes of flavonoids. As can be seen, flavonoids present different properties such as molecular weight and solubility which determines their reactivity and interaction with targets and finally defines their physiological activities. Within each class of flavonoids, the structure of compounds can differ considerably according to the type of substitution(s) they have undergone include glycosilation (D-glucose, lignin, arabinose, glucorhamnose, L-rhamnose, galactose), methylation, malonylation, hydrogenation, hydroxylation, and sulphation (Cook and Samman, 1996). In nature flavonoids are mainly found as flavonoid glycosides (e.g. rutin and quercitrin), whereas only occasionally occur in the form of aglycones (Cook and Samman, 1996; Heim et al., 2002; Ross and Kasum, 2002).

(Heijnen and van der Sluijs, 2015). The third category of platelet granules are the lysosomes which posses an intraluminal acidic pH with various active hydrolytic enzymes against a series of targets such as components of the extracellular matrix (Ciferri et al., 2000; Rendu and Brohard-Bohn, 2001). For over 120 years, many scientists have focused the research on the key role of platelets as essential mediators in hemostasis and its relation with pathological thrombosis. This hemostatic roles of platelets are essential for the maintenance of the integrity and homeostasy of the high-pressured circulatory system (Furie and Furie, 2008). In the presence of a distruption of the endothelial monolayer, the exposure of sub-endothelial matrix proteins rapidly engage specific receptors on the platelet surface, leading to biochemical and morphological changes in platelets, which undergo aggregation and support a localized activation of the coagulation cascade. Upon platelet stimulation, granules release their contents into the extracellular environment, contributing to platelet activation and thrombus formation (Andrews and Berndt, 2008). Several intracellular signalling pathways are involved in platelet activation once stimulated by contact to subendothelial collagen, thrombin, thromboxane A2 and ADP released by other activated platelets (Borst et al., 2013). In the last decade, scientific interests have been focused on non hemostatic activity of platelets which have been gradually better identified. Platelets were likely to play a key task in resistance and identification of pathogen invasion and in the mobilization of immune and inflammatory cells to the injured site and to enhance phagocytosis and pathogen killing (Vieira-de-Abreu et al., 2012; Li, 2008; Scull et al., 2010; Stephen et al., 2013). A multitude of immune-active molecules is secreted by platelets, including pro- and anti-inflammatory cytokines, chemokines, and anti-bacterial proteins (Golebiewska and Poole, 2015; Yeaman, 2010). Platelets can be, therefore, correlated to the pathogenesis of inflammatory disturbances including atherosclerosis, inflammatory bowel disease, rheumatoid arthritis, transplant rejection, or malaria infection (Senchenkova et al., 2015; Mazereeuw et al., 2013).

3. Main sources of flavonoids Flavonoids are found ubiquosly in plants (flowers, leaves, bark and seeds) and till the date more than 8000 different compounds have been identified. However, many more are still to be discovered as well as their metabolic pathways (Ferreyra et al., 2012). Their major role in plants is to protect against ultraviolet radiation, oxidative cell injury, pathogens and predactors, control the transport of auxins and to add pollinization due to the coloration of flowers which attracts pollinators (Cook and Samman, 1996; Ferreyra et al., 2012; Heim et al., 2002). The amount of flavonoids present in fruits and vegetables may present differences due to species variety, part of the plant, edaphoclimatic conditions, type of cultivation, degree of ripeness (Kozlowska and Szostak-Wegierek, 2014; Ross and Kasum, 2002). In foods, flavonoids are responsible for colour, taste, protection of enzymes and vitaminic compounds and avoiding of lipid peroxidation (Kumar and Pandey, 2013). Moreover, culinary process and storage can also greatly influence the content of flavonoids (Peterson and Dwyer, 1998). All together make difficult to estimate the daily dietary intake of flavonoids because of the high number of food sources, their wide distribution in plants and the diverse patterns of consumption (Kumar and Pandey, 2013; Trischitta and Faggio, 2006). The dietary souces of some classes of flavonoids based on the USDA database for the flavonoid content of selected foods are resumed in Table 2 (Bhagwat et al., 2011). Favan-3-ols are generally found in tea, flavanones in citrus fruits, anthocyanidins in colored fruits such as cherries and grapes (Cook and Samman, 1996). Anthocyanidins are responsible for the blue, red and violet colour of fruits like plums and berries and their content increase with maturation (Erlund, 2004) (Peterson and Dwyer, 1998). Isoflavonoids (isoflavones, isoflavanones and isoflavonols) are generally found in legumes Bhagwat et al. (2008) (Peterson and Dwyer, 1998). Aromatic plants are also an excellent source of flavonoids (Costa et al., 2015; Ribeiro-Santos et al., 2015).

2. Chemistry of flavonoids Flavonoids are polyphenolic compounds, characterized by the flavan nucleus. Fig. 1 presents the general structure of flavonoids showing the three phenolic rings. A, B and C are the three phenolic rings. A ring is condensed with C ring, which has B ring in the 2position as a substitute. Different classes of flavonoids differ on the level of oxidation and substituents of C ring. Individual flavonoids within each class differ on the number and type of substitutions of different groups of rings A and B (Kumar and Pandey, 2013). Although flavonoids have low molecular weight (Heim et al., 2002), proanthocyanidins can achieve high molecular weights, up to 17 flavanol units. In plants, flavonoids are sinthetized through the phenylpropanoid pathway, which transforms phenylalanine into 4-coumaroyl coenzyme A and then this enters in the flavonoid pathway. The phenylpropanoid

4. Bioavailability and metabolism of flavonoids Since dietary flavonoids have been reported to exert beneficial health effects, the understanding of their pharmacokinetics and metabolism is essential to determine the potential therapeutic uses.

Fig. 1. Basic structure of flavonoids.

92

European Journal of Pharmacology 807 (2017) 91–101

C. Faggio et al.

hydrolysis of some flavonoid glycosides, can occur in oral cavity, not being limited to colon. Then, the resultant products will be absorbed into human circulation or degraded in the colon. Once the flavonoid aglycones are absorbed by the intestinal cells, they will undergo an important metabolism by the action of diverse enzymes - mainly transferases - from the phase II metabolism rather than by the phase I metabolism cytochrome P450s (Chen et al., 2014). The main conjugated metabolites of flavonoids are glucuronidated forms followed by sulphated and methylated forms. Glucuronidation of flavonoids is catalysed by diverse isoforms of UDP-glucuronosyltransferases (UGT) which are found both in the intestine and the liver and are the responsible for about 80% of the metabolic pathway of flavonoids (Wu et al., 2011). Sulfation of flavonoids is mediated by sulfotransferases (SULTs) which show an elevated preference for sulfation at the C7–OH moiety (Meng et al., 2012). The methylation reaction is mediated by catechol-O-methyltransferases and is mainly produced in chatecholic flavonoids (Chen et al., 2013; Wright et al., 2010a, 2010b). Similar to flavonoid glycosides the glucuronidated and sulphated forms are too hydrophilic to cross biological membranes by passive diffusion whereas methylated forms are more lipophilic and can better cross the membranes. Both the intestine and liver contain a wide array of membrane pumps or efflux transporters called ATP-binding cassette (ABC) transporters (Gonzales et al., 2015). Some of these transporters have been reported to transport conjugated flavonoids across cellular membranes (Wei et al., 2013; Yang et al., 2012). Diverse transporters have different location in the apical and basolateral sides of cells being responsible to the efflux of flavonoids into the blood but also into the bile or returning them to the intestine lumen (Chen et al., 2014;

However, there is still controversy regarding the absorption of flavonoids. Most of the bioavailability studies focus on the individual flavonoid taken from pharmaceutical formulations instead of focusing of the bioavailability of dietary flavonoids, difficulting the extrapolation to the results of dietary flavonoids (Cook and Samman, 1996; Freese et al., 2002; Erlund et al., 2003). Bioavailability depends on the flavonoid, source (food or pharmaceutical formulation), gender, interindividual variations (including physiological and molecular factors), composition and activity of gastrointestinal microflora, mechanisms of absorption and biotransformation (Erlund, 2004; Kumar and Pandey, 2013). Most flavonoids are ingested in the form of O-glycosides so they are too hydrophilic to be absorbed directly by passive transport in the intestine. First, these glycosides must be hydrolyzed by intestinal hydrolases or the intestinal microflora in order to be subsequently absorbed (Chuankhayan et al., 2007; Hanske et al., 2010; Heim et al., 2002). On the contrary, the aglycone forms are more hydrophobic and therefore are better absorbed by enterocytes through passive diffusion. Human small intestine expresses a lactase-phlorizin hydrolase, a βhydrolase with a catalytic lactase site capable to hydrolyze flavonoid βglucosides (Day et al., 2000; Mitchelmore et al., 2000). The hydrolysis in the small intestine allows a fast absorption of flavonoid β-glucosides (Németh et al., 2003). The remaining flavonoids glycosides, containing in their structure other sugars such as rhamnose or rhamnoglucose, should be attacked by hydrolases of the intestinal microflora (Simons et al., 2005; Amaretti et al., 2015). In addition to the hydrolases, human microflora contains many other enzymes which can degrade almost all flavonoids into more simple molecules (Braune et al., 2001; Kim et al., 1998). A study conducted by Walle (2004) also revealed that Table 1 Physical-chemical properties of some representative flavonoids.

(continued on next page)

93

European Journal of Pharmacology 807 (2017) 91–101

C. Faggio et al.

Table 1 (continued)

(continued on next page)

94

European Journal of Pharmacology 807 (2017) 91–101

C. Faggio et al.

Table 1 (continued)

(a) Data from ChemIDplus advanced (http://chem.sis.nlm.nih.gov/chemidplus/; (b) ChemSpider (www.chemspider.com).

Alvarez et al., 2010). All together indicates that most flavonoids suffer from entero-enteric and entero-hepatic cycle with a very complex metabolism. Flavonoids returned to small instestine, the secreted with bile and those not absorbed by small instestine, undergo degradation by enzymes (β-glucosidase, α-rhamnosidade) produced by colon microflora and originate phenolic acids, such as hydroxyphenylacetic acids and hydroxyphenylpropionic acids (Erlund, 2004; Hollman, 2004). These can be absorbed and found in plasma and urine.

2014). The activation of phospholipase C (PLC) via G protein, results in the synthesis of inositol 1,4,5- triphosphate (IP3) and diacylglycerol (DG) which plays a central role in platelet aggregation and contributes to elevate the levels of free Ca2+([Ca2+]i) in the cytosol (Brass and Joseph, 1985; Williamson et al., 1985). Cytosolic Ca2+ activates PLA2 in platelets, promoting the release of arachidonic acid (AA), which in turn is transformed into thromboxane A2 (TXA2) by cyclooxygenase-1 (COX-1), which subsequently amplifies platelet activation and aggregation. Flavonoids have demonstrated several anticarcinogenic and antineurodegenerative effects (Fabiani et al., 2002; Fini et al., 2008; Heiss et al., 2005; Mantena et al., 2006; Corona et al., 2009; Wang et al., 2000; Piao et al., 2006; Guo et al., 2013; Karuppagounder et al., 2013). However, the beneficial effects of flavonoids on the cardiovascular system are the most extensively studied due to their effects on lipid metabolism (Jeong et al., 2005; Zern et al., 2005; Fuhrman et al., 2005; Hubbard et al., 2003), the capability to reduce cell adhesion (Ludwig et al., 2004) and potentiate the endothelial and vascular function (Hallund et al., 2006; Lemos et al., 1999; Almeida et al., 2002; Côrtes et al., 2001; Gonçalves et al., 2009; Rezende et al., 2009, 2004). Moreover, evidences have been reported that some flavonoids possess antiplatelet effects (Guerrero et al., 2005; Kim and Yun-Choi, 2008; Mower et al., 1984; Tzeng et al., 1991). Although numerous mechanisms have been reported such as inhibition of phospholipase A2, phosphodiesterases and diverse protein

5. Flavonoids and platelet aggregation Platelets are crucial not only in the hemostasis but also in wound healing processes (Nurden et al., 2007). Nevertheless, diverse studies evidenced that excessive platelet activation is directly related to many pathological disorders such as diabetes mellitus, hypertension or vascular diseases (El Haouari et al., 2007, El Haouari and Rosado, 2008; Elwood et al., 1990), and also participates in the progression of thrombotic diseases, and the occurrence of cardiovascular disorders (Tran and Anand, 2004). Platelet adhesion and activation are critically important for the progression of acute thrombotic occlusion in areas suffering from atherosclerotic plaque rupture, the main pathophysiological mechanism underlying ischemic disorders, such as stroke and myocardial infarction (Borst et al., 2013; Fauci, 2008; Watson et al., 2005). Platelet activation leading to thrombus formation decisively depends on an elevation of cytosolic Ca2+ levels ([Ca2+]i) (Borst et al., 95

European Journal of Pharmacology 807 (2017) 91–101

C. Faggio et al.

Table 2 Dietary sources of flavonoides (Bhagwat et al., 2011). Class of flavonoids

Food groups

Example of foods (mean values, content in mg/100 g edible portion)

Flavones

Spices and herbs

Oregano (3.57), peppermint (18.1), rosemary (2.55), sage (17.9), celery seed (841), marjoram (3.50), parsley (4523), tarragon (1.00), thyme (47.75) Olive oil (0.21) Apples (0.01–0.12), blueberries (0.20), cranberries (0.03), grapefruit (pink and red, 0.60), red grapes (1.30), juniper berries (58.66), kiwifruit (0.74), kumquats (21.87), mango (0.03), melon (0.64), lemon (1.90), olives (0.56), orange (0.19), papaya (0.03), persimmons (0.14), plum (0.02-red; 0.60- black diamond), watermelon (0.46) Annual saw-thistle (10.3), green beans (0.13), beets (0.37), broccoli (0.33–0.80), Brussels sprouts (0.33–0.50), cabbage (0.87), carrots (0.12), cauliflower (0.24–0.29), celeriac (2.41), celery (3.90–58.9), chicory greens (2.85), chives (0.15), corn poppy (0.20), eggplant (0.03), fennel (0.10), hartwort (0.60), hawthorn (0.40), horseradish (0.90), lettuce (0.05–0.95), lotus root (0.36), onions (0.01–0.40), peas (cooked, 0.41), peppers (0.07–3.57), perilla leaves (0.39), pumpkin (1.63), queen Anne's lace (160.1), radicchio (37.98), rutabagas (3.85), spinach (0.74), sweet potato (0.03–0.17), tomatoes (0.01–2.86), vinespinach (62.2), water spinach (0.05), watercress (0.03), yardlong bean (0.03) Red wine (0.04–0.17), green tea (0.30) Sorghum (2.99–6.47)

Fats and oils Fruits

Vegetables and vegetables products

Beverages Cereal grains and pasta Flavonols

Dairy products Spices and herbs Fruits

Vegetables and vegetables products

Nuts and seeds Beverages Legumes and legume products Flavanones

Flavan-3-ols

Spices and herbs Fruits Vegetables and vegetables products Nuts and seeds, Beverages Cereal grains and pasta

Oregano (0–457.3), peppermint (41.1), rosemary (24.9) Grapefruit (22.0–54.5), lemon (49.8), orange (29.0–42.6), pummel (33.1), strawberries (0.26), yuzu (53.5) Brussels sprouts (cooked −1.94; raw- 3.29), fennel bulb (1.08), tomatoes (0.68–3.19)

Dairy products Fruits

Milk chocolate (1.08) Apple (6.64–12.3), apricot (8.41), avocado (0.52), banana (6.12), blackberries (42.5), blueberries (6.69–124.1), cherries (4.13–9.75), cloudberries (1.3), cranberries (6.47), currants (0.30–2.78), custard-apple (6.25), figs (2.09), gooseberries (2.11), grapes (2.03–21.63), jujube (3.52), kiwifruit (0.37–0.64), mango (1.72), medlar (0.79), nectarines (5.52–10.6), peach (1.87–16.3), pear (1.88–4.81), persimmons (0.80), plum (7.58–33.5), pomegranates (0.81), quince (1.42), raisins (0.52), raspberries (5.83), rhubarbs (2.35–3.28), star apple (1.65), strawberries (4.60) Chard (0.20–6.7), onions (0–0.08), peas (0–0.02)

Vegetables and vegetables products Nuts and seeds Beverages Legumes and Legume products Anthocyanidins

Milk chocolate (0.17) Capers (303.9–493.0), chives (17.1), dill weed (112.7), oregano (9.40), parsley (331.2), saffron (205.5), tarragon (36.0) Acerola (5.79), apple (0.42–3.87), apricot (2.36), arctic bramble berries (10.65), banana (0.18), bayberries (8.01), bilberry (4.13), blackberries (4.52), blueberries (7.5–19.7), bog whortleberries (25.0), cashew apple (3.38), cherries (2.43–28.6), chokeberry (18.9), cloudberries (0.57), cranberries (6.91–21.6), crowberries (10.1), currants (1.69– 11.5), dates (0.93), elderberries (32.8), figs (5.47), goji berry (31.2), goopseberries (2.11), grapefruit (0.35–0.90), grapes (1.05–2.39), guava (1.00–1.20), jujube (1.26), juniper berries (42.8–46.6), kiwifruit (1.07), lemon (1.67), lime (0.40), lingonberries (13.7), mango (0.11), melon (0.08), mulberries (2.47), nectarine (0.12–0.69), orange (0.22–0.73), papaya (0.03), peach (0.45–0.88), pear (1.14), persimmons (1.06), pineapple (0.15), plum (0.90– 12.5), prickly pear (5.69), raisins (0.27–5.11), raspberries (1.11–1.14), sea buckthorn berry (45.9), star apple (0.34), strawberries (1.3–1.65), watermelon (0.45) Alfafa seeds (1.70), amaranth leaves (1.43), annual saw-thistle (24.1), arugula (47.1), asparagus (raw- 21.1; cooked 15.2), bay leaves (8.01), beans (1.51–3.65), beets (0.13), broadbeans (4.60), broccoli (1.05–11.2), Brussels sprouts (2.78–5.24), cabbage (0.05–22.5), carrots (0.49), catsup (0.87), cauliflower (frozen and boiled-0.58; raw-0.90), celeriac (0.18), celery (0.61), chard (1.55–18.9), chicory greens (8.94), chives (21.5), collards (11.3), coriander (52.9), corn poppy (30.8), cowpeas (5.50), cress (14.0), crown daisy (0.18), cucumber (0.17), dock (102.2), drumstick (23.0), eggplant (0.04–0.08), endive (10.1), fennel (0.23–84.4), garlic chives (2.24), garlic (3.61), ginger (0.19–33.6), gourd (0.16), hartwort (24.1–38.9), horseradish (1.86), kale (0.08–93.0), kohlrabi (2.83), leeks (2.98), lettuce (1.63–7.63), lotus root (1.79), lovage (177), mizuna (18.4), mung beans (0.48), mustard greens (0.84–63.3), nalta jute (30.1), okra (21.0), onions (3.63–46.7), pako fern (0.63), peas (0.11–0.22), peppers (0.09–50.63), perilla leaves (0.96), potatoes (0.49–1.50), purslane (4.24), queen Anne's lace (1.70), radicchio (31.5), radish seeds (78.1), radishes (0.34–0.86), rocket (68.8), rutabagas (2.5), sauerkraut (0.05), soybeans (1.26), spinach (10.7), squash (0.47–0.66), sweet potato leaves (13.7–23.5), taro leaves (0.05–0.42), taro (0.23–2.87), tomatoes (0.03–0.8), tree spinach (5.41–7.11), turmeric (6.96), turnip greens (12.6), water spinach (1.92), watercress (0.90–53.0), yam (0– 0.25), yardlong (5.83) Almonds (3.15), Chia seeds (30.7), pistachio (1.46) Beer (0.85), champagne (0.01), wine (0.01–1.94), cocoa mix (2.03), coffee (0.10), tea (1.17–4.82) Beans (2.58–26.0), broadbeans (0.9), carob fiber (117.5), carob flour (46.0), carob kibbles (15.9), cowpeas (21.9), tofu (1.19)

Fruits

Almonds (0.68) Wine (0.78–2.40) Sorghum (1.96)

Almonds (4.47), cashew nuts (1.98), chestnuts (0.02), hazelnuts (5.25), pecans (16.0), pine nuts (0.49), pistachio (6.85) Beer (0.54), champagne (0.30), wine (1.32–18.4), cocoa mix (1.33–52.7), coffee (0.08), tea (black, green, fruit flavoured, white) (9.8–324.2) Beans (0.10–5.26), carob flour (190.27), lentils (0.49), marrowfat pea (9.97), peanuts (0.66), soybeans (37.4)

Acai (0.48–204.9), acerola (22.5), apple (0–5.0), arctic bramble berries (89.0), avocados (0.33), banana (7.39), bilberry (285.2), blackberries (100.6), blueberries (94.2–163.3), cashew apple (0.19), cedar bay cherry (27.8), cherries (2.42–34.5), chokeberry (349.8), cloudberries (1.70), cranberries (0.72–104.0), currants (1.0–157.8), dates (1.70), elderberries (485.28), figs (0.51), gooseberries (9.51), grapes, (47.7–120.1) guajiru (72.7), jambul (continued on next page)

96

European Journal of Pharmacology 807 (2017) 91–101

C. Faggio et al.

Table 2 (continued) Class of flavonoids

Food groups

Vegetables and vegetables products Nuts and seeds Beverages Legumes and Legume products Cereal grains and pasta

Example of foods (mean values, content in mg/100 g edible portion) (55.1), jostaberry (27.9), kiwifruit (1.65), lingonberries (40.2), mango (0.14), maqui (88.5), molucca raspberry (94.2), muntries (24.8), nectarines (0.74–2.13), peaches (0.97–1.92), pears (2.06), plum (0.30–558.2), raisins (0.05), raspberries (24.2–686.8), sea buckthorn berry (0.08), strawberries (20.6–27.0) Beans (0.06), cabbage (0.06–210.0), lettuce (red leaf), 3.14, onions (red, 9.56)), peas (0.08), peppers (0–752.7), radicchio (134.7), radish (63.13), purple sweet potato (11.5), taro leaves (0.06), yardlong bean (1.14) Almonds (2.46), hazelnuts (6.71), pecans, (18.0) pistachio (7.33), walnuts (2.71) Wine (0.06–153.0) Beans (2.74–44.5), cowpeas (262.5) Purple wheat (25.9)

Note 1: Flavonols: Quercetin, Kaempferol, Myricetin, Isorhamnetin; Flavones: Luteolin, Apigenin; Flavanones: Hesperetin, Naringenin, Eriodictyol; Flavan-3-ols: (+)-Catechin, (+)-Gallocatechin, (-)-Epicatechin, (-)-Epigallocatechin, (-)-Epicatechin 3-gallate, (-)-Epigallocatechin 3-gallate, Theaflavin, Theaflavin 3-gallate, Theaflavin 3′-gallate, Theaflavin 3,3′ digallate, Thearubigins; Anthocyanidins: Cyanidin, Delphinidin, Malvidin, Pelargonidin, Peonidin, Petunidin. Note 2: when the content is presented as a range of values, it correspond to the mean values of two or more independent groups (e.g. from different varieties, processed differently).

1979)), degranulation and integrin signalling mediated by αIIbβ 3 (Shattil and Brass, 1987), inhibition of platelet granule secretion (platelet secretion is decisively implicated in platelet aggregation and thrombus development (Schmidt et al., 2013)) increase of cyclic nucleotides (in platelets, it is established that cAMP- and cGMPdependent effects blocks agonist-induced augments in cytosolic Ca2+ levels and granule secretion (Aszódi et al., 1999)), inhibition of formation of thromboxanes (TXA2 is a potent stimulator and effective vasoconstrictor synthesized by platelets during the aggregation process (Bunting et al., 1983; Cho et al., 2006)), and decrease in phosphoinositide breakdown and protein kinase C inductors. Other mechanisms can also be involved including inhibition of phospholipase C, plateletactivating factor (PAF) or collagen-receptor antagonism and GPIIb-IIIa activation (platelet inside-out signalling to integrin αIIbβ3 is crucial for the regulation of its conformation to amplify the affinity for plasma fibrinogen and von Willebrand factor (vWF) binding and subsequent platelet aggregation (Calderwood, 2004)), and reduction of protein tyrosine phosphorylation. Many flavonoids have been evidenced to inhibit numerous enzymes, associated with the regulation of normal cellular functioning including phospholipase As (PLAs), tyrosine kinases, phosphodiesterases, lipoxygenases and COX isoforms (Benavente-Garcia and Castillo, 2008; Benavente-García et al., 1997; Wright et al., 2013). Nitric oxide (NO) is another interesting point in which flavonoids can modulate platelet aggregation. Treatment of platelets with quercetin and catechin increased the production NO, followed by a down regulation in the expression of glycoprotein IIb/ IIIa complex (Pignatelli et al., 2006). Similarly, other investigation reported that treatment with flavonoids such as curcuminoids or substituted stilbenes increased NO levels in agonist-activated platelets (Maheswaraiah et al., 2015; Messina et al., 2015). However, in another study NO did not mediate the inhibitory effect of rutin on a platelet aggregation (Sheu et al. 2004). Moreover, flavonoids as phenolic compounds have the antioxidant effects with notable capability to scavenge free radicals (Bors et al., 1990). It has been evidenced that collagen-induced platelet aggregation is directly associated with an oxidative burst which participates in the platelet activation via calcium increase and activation of the inositol pathway (Pignatelli et al., 1998). Diverse flavonoids such as quercetin and catechin significantly reduced the oxidative burst induced by collagen and, consequently, inhibited platelet aggregation (Pignatelli et al., 2000). In this way, it was reported that some flavonoids such as kaempferol inhibit NADPH oxidase, thus, reducing the production of reactive oxygen species (Wang et al., 2015). In contrast to the in vitro and ex vivo studies, clinical trials are still scarce and performed with a low number of patients. Moreover, the obtained results are variable, finding beneficial effects of flavonoids in reducing platelet reactivity but also the absence of positive effects. In a pilot study the effects of one week of dark chocolate intake (700 mg flavonoids/day) was analyzed in 28 healthy volunteers (Hamed et al., 2008). The ADP- and AA-induced platelet activation as well as

kinases (Beretz et al., 1982; Guerrero et al., 2007; Hubbard et al., 2003; Lindahl and Tagesson, 1997), the specific underlying mechanism, which appears to be common for most of flavonoids compounds seems to imply the inhibition platelet aggregation through the arachidonic acid-based pathway (Fig. 2) (Mladěnka et al., 2010; Navarro-Nunez et al., 2009). Certainly, flavonoids are capable to reduce the aggregation induced by different stimulators. Certain flavonoids, rather at higher concentrations, may avoid the aggregation provoked by multiple G protein coupled receptor agonists ADP or thrombin (Tzeng et al., 1991; Luzak et al., 2017; Choi et al., 2016). Thrombin and ADP induce platelet activation through a conformational change of αIIbβ3 integrin increasing the affinity state to fibrinogen (Rink, 1990). Particularly, flavonoids appear to act as strong inhibitors of aggregation induced by AA, but also by collagen, which is recognized to play the central role in phase 1 of pathophysiological platelet aggregation (Jackson, 2007). The underlying mechanisms in which flavonoids block the platelet activation are complex and depend on the specific flavonoid. In this sense, a significant attenuation in the phosphorylation of one or more pathways has been reported in stimulated platelets. Specifically, flavonoid inhibition seems to be mainly mediated by phosphoinositide 3-kinase (PI3K)/PKB (AKT) but also by extracellular signal-regulated kinase (ERK) 1/2, p38 and c-Jun N-terminal kinase (JNK) 1/2 pathways (Tian, 2016; Choi et al., 2015; Hao et al., 2015; Liang et al., 2015). Another effect of flavonoids seems to be mediated by antagonism on TXA2 receptors (Guerrero et al., 2005). TXA2 is the main cyclooxygenase product of arachidonic acid in platelets and acts via a membrane surface receptor to induce platelet aggregation. A study reported that genistein, daidzein and equol, but not the glucuronidated forms can bind to the TXA2, suggesting that the presence of glucoside residues may increase the flavonoid size avoiding the binding capability (Muñoz et al., 2009). Another study assessing the inhibitory effects of 20 flavonoids with different structural characteristics concluded that the flavone apigenin and the isoflavone genistein are the structures evidencing the highest binding capability (Navarro-Nunez et al., 2009). In addition, several data suggested that flavonoids decrease levels of TXA2 in an indirect way mainly related to inhibition of COX-1 (Corvazier and Maclouf, 1985). Inhibition of COX-1 specifically depends on an isoflavone ring containing a 7-hydroxyl group, whereas to act as an antagonist at thromboxane A2 receptors, this group is not absolutely necessary, since its blockade in apigenin-7- glucoside did not eradicate the inhibitory activity. As an alternative, the presence of a glucose residue at position 7 in isoflavones reduced the effect. The position of ring B (isoflavones vs. corresponding flavones) is significant for COX-1 inhibition but not for antagonism at thromboxane receptors. The antiplatelet activity of the plant extracts is also associated to other mechanisms, such as inhibition of cytoplasmic Ca2+ augment (an increase in the levels of intracellular calcium in platelets is necessary for the modulation of thrombus development, reorganization of the actin cytoskeleton required for shape change (Hathaway and Adelstein, 97

European Journal of Pharmacology 807 (2017) 91–101

C. Faggio et al.

Fig. 2. Mechanism of inhibition of platelet aggregation by flavonoids. Flavonoids inhibit the signalling pathways initiated by thromboxane A2 (TXA2), ADP, thrombin and collagen receptors and αIIbβ3 integrin. Flavonoids also block the thrombin and ADP induced conformational change of αIIbβ3 integrin. Flavonoids prevent platelet aggregation inhibiting intracellular signalling pathways, the enzymatic activities of phospholipase C, phospholipase A2 (PLA2) and cyclooxygenase 1 (COX-1), and reducing the oxidative burst. Flavonoids also prevent the decrease in nitric oxide production associated to platelet aggregation.

2000 mg of cocoa flavonols during 12 weeks (Ottaviani et al., 2015).

activated glycoprotein IIb/IIIa were significantly reduced by dark chocolate. A non-randomized study with 65 healthy subjects investigated the effects of 18.75 mg of flavonoid rich dark chocolate added to 100 mg oral aspirin (Zubair et al., 2011). The results showed an increased anti platelet effects of aspirin when consumed in conjunction with chocolate. The consumption of dark chocolate enriched with flavan-3-ol, but also white chocolate, reduced platelet aggregation in an intervention trial with 42 healthy subjects with differentiate results depending on the gender (Ostertag et al., 2013). Low and high flavanol apple purees containing 25 or 100 mg epicatechin diminished epinephrine- and ADP -induced integrin-β3 expression and increased plasma NO metabolites (Gasper et al., 2014). The supplementation with dark chocolate containing high and low flavanol levels (1064 and 88 mg flavanols/day, respectively) during 6 weeks, reduced platelet responsiveness to ADP and to thrombin receptor activator peptide but not to the thromboxane analogue U46619 or collagen (Rull et al., 2015). The authors suggest that the effects are due to the theobromine content rather than to flavonols. However, in many studies the in vivo antiplatelet effects of flavonoids are lower or without evident effects than expected from previous in vitro assays. In fact, caffeic acid has shown notable anti-aggregation effects whereas a clinical intervention with caffeine-free chicory coffee, rich in caffeic acid did not report clear results (Hung et al., 2005; Schumacher et al., 2011). A double-blind randomized intervention supplying with monomeric and oligomeric flavonoids from grape seeds (200 mg/day) to 28 male smokers did not find any effect on platelet aggregation (Weseler et al., 2011). Nerveless, integrating all measured data into a general health index, the supplementation induced a significant improvement in vascular health. The in vivo effects of wine polyphenols using de-alcoholised wine were performed in 21 post-menopausal women (Giovannelli et al., 2011). Platelet aggregation measured using epinephrine-collagen and ADPcollagene as activators did not evidence significant differences. In another intervention study no effects were reported in subjects with stage 1 hypertension in platelet aggregation after 8 weeks of grape seed extract supplementation (Ras et al., 2013). In addition, lack of differences was also found in healthy subjects with a daily intake of

6. Conclusion and future prospects The progressive increase in the prevalence of cardiovascular diseases requires new efforts in the development of new therapeutic agents. Flavonoids are secondary metabolites very abundant in plants and consequently important constituents of the human diet. The available data suggests a potential therapeutic effect of several flavonoids via modulation platelet aggregation against cardiovascular diseases. Diverse mechanisms of action has been reported including the inhibition of the arachidonic acid pathway, the suppression of cytoplasmic Ca2+ increase, the blockage of the degranulation and integrin αIIbβ 3-mediated signalling, the inhibition of platelet granule secretion and inhibition of thromboxane formation, among others. Even though in vitro and ex vivo data seem to sustain epidemiological studies, the way to the ultimate confirmation of a positive effect of flavonoids on platelet aggregation and their effect in pathological disorders requires supplementary investigations. The large number of different flavonoids together with the complex secondary metabolism they are subjected to and the fact that clinical trials are scarce and with few patients makes it difficult to obtain conclusive results. In addition, the general bioavailability of these compounds is poor and depends on many factors including the type of the flavonoid, source and mechanisms of absorption and biotransformation. Additional well controlled intervention studies with higher number of subjects are compulsory for improved understanding of the connection between plant flavonoids and platelet function may help to the development of better strategies to treat cardiovascular disorder. Altogether, the results about the antiplatelet aggregation of flavonoids are promising but deeper studies about the bioavailability, molecular mechanism of action, efficacy doses and toxicity are mandatory before these compounds could be used and pharmacological agents. 98

European Journal of Pharmacology 807 (2017) 91–101

C. Faggio et al.

Corvazier, E., Maclouf, J., 1985. Interference of some flavonoids and non-steroidal antiinflammatory drugs with oxidative metabolism of arachidonic acid by human platelets and neutrophils. Biochim. Biophys. Acta (BBA)-Lipids Lipid Metab. 835, 315–321. Costa, D.C., Costa, H., Albuquerque, T.G., Ramos, F., Castilho, M.C., Sanches-Silva, A., 2015. Advances in phenolic compounds analysis of aromatic plants and their potential applications. Trends Food Sci. Technol. 45, 336–354. Day, A.J., Cañada, F.J., Díaz, J.C., Kroon, P.A., Mclauchlan, R., Faulds, C.B., Plumb, G.W., Morgan, M.R., Williamson, G., 2000. Dietary flavonoid and isoflavone glycosides are hydrolysed by the lactase site of lactase phlorizin hydrolase. FEBS Lett. 468 (2–3), 166–170. Ebbeling, L., Robertson, C., McNicol, A., Gerrard, J.M., 1992. Rapid ultrastructural changes in the dense tubular system following platelet activation. Blood 80, 718–723. El Haouari, M., Rosado, J.A., 2008. Platelet signalling abnormalities in patients with type 2 diabetes mellitus: a review. Blood Cells Mol. Dis. 41, 119–123. El Haouari, M., López, J.J., Mekhfi, H., Rosado, J.A., Salido, G.M., 2007. Antiaggregant effects of Arbutus unedo extracts in human platelets. J. Ethnopharmacol. 113, 325–331. Elwood, P.C., Beswick, A.D., Sharp, D.S., Yarnell, J., Rogers, S., Renaud, S., 1990. Whole blood impedance platelet aggregometry and ischemic heart disease. The Caerphilly Collaborative Heart Disease Study. Arteriosclerosis 10, 1032–1036. Erlund, I., 2004. Review of the flavonoids quercetin, hesperetin, and naringenin. dietary sources, bioactivities, bioavailability, and epidemiology. Nutr. Res. 24, 851–874. Erlund, I., Marniemi, J., Hakala, P., Alfthan, G., Meririnne, E., Aro, A., 2003. Consumption of black currants, lingonberries and bilberries increases serum quercetin concentrations. Eur. J. Clin. Nutr. 57, 37–42. Fabiani, R., De Bartolomeo, A., Rosignoli, P., Servili, M., Montedoro, G., Morozzi, G., 2002. Cancer chemoprevention by hydroxytyrosol isolated from virgin olive oil through G1 cell cycle arrest and apoptosis. Eur. J. Cancer Prev. 11, 351–358. Fauci, A.S., 2008. Harrison's principles of internal medicine. Medical Publishing Division, McGraw-Hill. Ferreyra, M.L.F., Rius, S.P., Casati, P., 2012. Flavonoids: biosynthesis, biological functions, and biotechnological applications. Front. Plant Sci. 3, 222. Fini, L., Hotchkiss, E., Fogliano, V., Graziani, G., Romano, M., Edward, B., Qin, H., Selgrad, M., Boland, C.R., Ricciardiello, L., 2008. Chemopreventive properties of pinoresinol-rich olive oil involve a selective activation of the ATM–p53 cascade in colon cancer cell lines. Carcinogenesis 29, 139–146. Freese, R., Alfthan, G., Jauhiainen, M., Basu, S., Erlund, I., Salminen, I., Aro, A., Mutanen, M., 2002. High intakes of vegetables, berries, and apples combined with a high intake of linoleic or oleic acid only slightly affect markers of lipid peroxidation and lipoprotein metabolism in healthy subjects. Am. J. Clin. Nutr. 76, 950–960. Fuhrman, B., Volkova, N., Coleman, R., Aviram, M., 2005. Grape powder polyphenols attenuate atherosclerosis development in apolipoprotein E deficient (E0) mice and reduce macrophage atherogenicity. J. Nutr. 135, 722–728. Furie, B., Furie, B.C., 2008. Mechanisms of thrombus formation. N. Engl. J. Med. 359, 938–949. Gasper, A., Hollands, W., Casgrain, A., Saha, S., Teucher, B., Dainty, J.R., Venema, D.P., Hollman, P.C., Rein, M.J., Nelson, R., Williamson, G., Kroon, P.A., 2014. Consumption of both low and high (-)-epicatechin apple puree attenuates platelet reactivity and increases plasma concentrations of nitric oxide metabolites: a randomized controlled trial. Arch. Biochem. Biophys. 559, 29–37. George, J.N., 2000. Platelets. Lancet 355, 1531–1539. Giovannelli, L., Pitozzi, V., Luceri, C., Giannini, L., Toti, S., Salvini, S., Sera, F., Souquet, J.M., Cheynier, V., Sofi, F., Mannini, L., Gori, A.M., Abbate, R., Palli, D., Dolara, P., 2011. Effects of de-alcoholised wines with different polyphenol content on DNA oxidative damage, gene expression of peripheral lymphocytes, and haemorheology: an intervention study in post-menopausal women. Eur. J. Nutr. 50 (1), 19–29. Golebiewska, E.M., Poole, A.W., 2015. Platelet secretion: from haemostasis to wound healing and beyond. Blood Rev. 29, 153–162. Gonçalves, R.L., Lugnier, C., Keravis, T., Lopes, M.J., Fantini, F.A., Schmitt, M., Cortes, S.F., Lemos, V.S., 2009. The flavonoid dioclein is a selective inhibitor of cyclic nucleotide phosphodiesterase type 1 (PDE1) and a cGMP-dependent protein kinase (PKG) vasorelaxant in human vascular tissue. Eur. J. Pharmacol. 620, 78–83. Gonzales, G.B., Smagghe, G., Grootaert, C., Zotti, M., Raes, K., Van Camp, J., 2015. Flavonoid interactions during digestion, absorption, distribution and metabolism: a sequential structure-activity/property relationship-based approach in the study of bioavailability and bioactivity. Drug Metab. Rev. 47 (2), 175–190. Guerrero, J.A., Lozano, M.L., Castillo, J., Benavente-García, O., Vicente, V., Rivera, J., 2005. Flavonoids inhibit platelet function through binding to the thromboxane A2 receptor. J. Thromb. Haemost. 3 (2), 369–376. Guerrero, J.A., Navarro‐Nuñez, L., Lozano, M.L., Martínez, C., Vicente, V., Gibbins, J.M., Rivera, J., 2007. Flavonoids inhibit the platelet TxA2 signalling pathway and antagonize TxA2 receptors (TP) in platelets and smooth muscle cells. Br. J. Clin. Pharmacol. 64, 133–144. Guo, D.J., Li, F., Yu, P.H.F., Chan, S.W., 2013. Neuroprotective effects of luteolin against apoptosis induced by 6-hydroxydopamine on rat pheochromocytoma PC12 cells. Pharm. Biol. 51, 190–196. Hallund, J., Bügel, S., Tholstrup, T., Ferrari, M., Talbot, D., Hall, W., Reimann, M., Williams, C., Wiinberg, N., 2006. Soya isoflavone-enriched cereal bars affect markers of endothelial function in postmenopausal women. Br. J. Nutr. 95, 1120–1126. Hamed, M.S., Gambert, S., Bliden, K.P., Bailon, O., Singla, A., Antonino, M.J., Hamed, F., Tantry, U.S., Gurbel, P.A., 2008. Dark chocolate effect on platelet activity, Creactive protein and lipid profile: a pilot study. South Med. J. 101 (12), 1203–1208. Hanske, L., Loh, G., Sczesny, S., Blaut, M., Braune, A., 2010. Recovery and metabolism of xanthohumol in germ-free and human microbiota-associated rats. Mol. Nutr. Food

Acknowledgements A. Sureda was supported by Spanish Ministry of Health and Consumer Affairs (CIBEROBN CB12/03/30038). A. Mocan acknowledges funding from UEFISCDI, Romania, project no. PNII-RU-TE2014-4-1247. References Almeida, A.P., Côrtes, S.F., Ferreira, A.J., Lemos, V.N.S., 2002. Increase on the coronary flow induced by dioclein in isolated rat heart. Life Sci. 70, 1121–1128. Alvarez, A.I., Real, R., Pérez, M., Mendoza, G., Prieto, J.G., Merino, G., 2010. Modulation of the activity of ABC transporters (P-glycoprotein, MRP2, BCRP) by flavonoids and drug response. J. Pharm. Sci. 99 (2), 598–617. Amaretti, A., Raimondi, S., Leonardi, A., Quartieri, A., Rossi, M., 2015. Hydrolysis of the rutinose-conjugates flavonoids rutin and hesperidin by the gut microbiota and bifidobacteria. Nutrients 7 (4), 2788–2800. Andrews, R.K., Berndt, M.C., 2008. Platelet adhesion: a game of catch and release. J. Clin. Investig. 118, 3009–3011. Aszódi, A., Pfeifer, A., Ahmad, M., Glauner, M., Zhou, X.H., Ny, L., Andersson, K.E., Kehrel, B., Offermanns, S., Fässler, R., 1999. The vasodilator‐stimulated phosphoprotein (VASP) is involved in cGMP‐and cAMP‐mediated inhibition of agonist‐induced platelet aggregation, but is dispensable for smooth muscle function. EMBO J. 18, 37–48. Benavente-Garcia, O., Castillo, J., 2008. Update on uses and properties of citrus flavonoids: new findings in anticancer, cardiovascular, and anti-inflammatory activity. J. Agric. Food Chem. 56, 6185–6205. Benavente-García, O., Castillo, J., Marin, F.R., Ortuño, A., Del Río, J.A., 1997. Uses and properties of citrus flavonoids. J. Agric. Food Chem. 45, 4505–4515. Beretz, A., Stierle, A., Anton, R., Cazenave, J.P., 1982. Role of cyclic AMP in the inhibition of human platelet aggregation by quercetin, a flavonoid that potentiates the effect of prostacyclin. Biochem. Pharmacol. 31, 3597–3600. Bhagwat, S., Haytowitz, D.B., Holden, J.M., 2008. USDA Database for the Isoflavone Content of Selected Foods Release 2.0.. United States Department of Agriculture, Nutrient Data Laboratory. Bhagwat, S., Haytowitz, D.B., Holden, J.M., 2011. USDA Database for the Flavonoid Content of Selected Foods, Release 3.1.. US Department of Agriculture, Beltsville. Bors, W., Heller, W., Michel, C., Saran, M., 1990. Flavonoids as antioxidant: determination of radical-scavenging efficiencies. Methods Enzymol. 186, 343–367. Borst, O., Walker, B., Münzer, P., Russo, A., Schmid, E., Faggio, C., Bigalke, B., Laufer, S., Gawaz, M., Lang, F., 2013. Skepinone-L, a novel potent and highly selective inhibitor of p38 MAP kinase, effectively impairs platelet activation and thrombus formation. Cell. Physiol. Biochem. 31, 914–924. Borst, O., Münzer, P., Schmid, E., Schmidt, E.M., Russo, A., Walker, B., Yang, W., Leibrock, C., Szteyn, K., Schmidt, S., 2014. 1, 25 (OH) 2 vitamin D3-dependent inhibition of platelet Ca2+ signaling and thrombus formation in klotho-deficient mice. FASEB J. 28, 2108–2119. Brass, L.F., Joseph, S., 1985. A role for inositol triphosphate in intracellular Ca2+ mobilization and granule secretion in platelets. J. Biol. Chem. 260, 15172–15179. Braune, A., Gütschow, M., Engst, W., Blaut, M., 2001. Degradation of quercetin and luteolin by Eubacterium ramulus. Appl. Environ. Microbiol. 67 (12), 5558–5567. Bunting, S., Moncada, S., Vane, J., 1983. The prostacyclin-thromboxane A2 balance: pathophysiological and therapeutic implications. Br. Med. Bull. 39, 271–276. Calderwood, D.A., 2004. Integrin activation. J. Cell Sci. 117, 657–666. Chen, Z., Zheng, S., Li, L., Jiang, H., 2014. Metabolism of flavonoids in human: a comprehensive review. Curr. Drug Metab. 15 (1), 48–61. Chen, Z.J., Dai, Y.Q., Kong, S.S., Song, F.F., Li, L.P., Ye, J.F., Wang, R.W., Zeng, S., Zhou, H., Jiang, H.D., 2013. Luteolin is a rare substrate of human catechol-Omethyltransferase favoring a para-methylation. Mol. Nutr. Food Res. 57 (5), 877–885. Cho, H.J., Cho, J.Y., Rhee, M.H., Lim, C.R., Park, H.J., 2006. Cordycepin (3 ‘‐ deoxyadenosine) inhibits human platelet aggregation induced by U46619, a TXA2 analogue. J. Pharm. Pharmacol. 58, 1677–1682. Choi, J.H., Park, S.E., Kim, S.J., Kim, S., 2015. Kaempferol inhibits thrombosis and platelet activation. Biochimie 115, 177–186. Choi, J.H., Kim, K.J., Kim, S., 2016. Comparative effect of quercetin and quercetin-3-Oβ-d-glucoside on fibrin polymers, blood clots, and in rodent models. J. Biochem. Mol. Toxicol. 30 (11), 548–558. Chuankhayan, P., Rimlumduan, T., Svasti, J., Cairns, J.R., 2007. Hydrolysis of soybean isoflavonoid glycosides by Dalbergia beta-glucosidases. J. Agric. Food Chem. 55 (6), 2407–2412. Ciferri, S., Emiliani, C., Guglielmini, G., Orlacchio, A., Nenci, G.G., Gresele, P., 2000. Platelets release their lysosomal content in vivo in humans upon activation. Thromb. Haemost. 83, 157–164. Cook, N., Samman, S., 1996. Flavonoids—chemistry, metabolism, cardioprotective effects, and dietary sources. J. Nutr. Biochem. 7, 66–76. Corona, G., Deiana, M., Incani, A., Vauzour, D., Dessì, M.A., Spencer, J.P., 2009. Hydroxytyrosol inhibits the proliferation of human colon adenocarcinoma cells through inhibition of ERK1/2 and cyclin D1. Mol. Nutr. Food Res. 53, 897–903. Côrtes, S.F., Rezende, B.A., Corriu, C., Medeiros, I.A., Teixeira, M.M., Lopes, M.J., Lemos, V.S., 2001. Pharmacological evidence for the activation of potassium channels as the mechanism involved in the hypotensive and vasorelaxant effect of dioclein in rat small resistance arteries. Br. J. Pharmacol. 133, 849–858.

99

European Journal of Pharmacology 807 (2017) 91–101

C. Faggio et al.

quercetin against sodium fluoride induced oxidative stress in rat's heart. Food Funct. 3, 437–441. Nabavi, S.F., Russo, G.L., Daglia, M., Nabavi, S.M., 2015. Role of quercetin as an alternative for obesity treatment: you are what you eat!. Food Chem. 179, 305–310. Navarro-Nunez, L., Castillo, J., Lozano, M.L., Martinez, C., Benavente-García, O., Vicente, V., Rivera, J., 2009. Thromboxane A2 receptor antagonism by flavonoids: structure− activity relationships. J. Agric. Food Chem. 57, 1589–1594. Németh, K., Plumb, G.W., Berrin, J.G., Juge, N., Jacob, R., Naim, H.Y., Williamson, G., Swallow, D.M., Kroon, P.A., 2003. Deglycosylation by small intestinal epithelial cell beta-glucosidases is a critical step in the absorption and metabolism of dietary flavonoid glycosides in humans. Eur. J. Nutr. 42 (1), 29–42. Nurden, A.T., Nurden, P., Sanchez, M., Andia, I., Anitua, E., 2007. Platelets and wound healing. Front. Biosci. 13, 3532–3548. Ostertag, L.M., Kroon, P.A., Wood, S., Horgan, G.W., Cienfuegos-Jovellanos, E., Saha, S., Duthie, G.G., de Roos, B., 2013. Flavan-3-ol-enriched dark chocolate and white chocolate improve acute measures of platelet function in a gender-specific way–a randomized-controlled human intervention trial. Mol. Nutr. Food Res. 57 (2), 191–202. Ottaviani, J.I., Balz, M., Kimball, J., Ensunsa, J.L., Fong, R., Momma, T.Y., Kwik-Uribe, C., Schroeter, H., Keen, C.L., 2015. Safety and efficacy of cocoa flavanol intake in healthy adults: a randomized, controlled, double-masked trial. Am. J. Clin. Nutr. 102 (6), 1425–1435. Peterson, J., Dwyer, J., 1998. Flavonoids: dietary occurrence and biochemical activity. Nutr. Res. 18, 1995–2018. Piao, M., Mori, D., Satoh, T., Sugita, Y., Tokunaga, O., 2006. Inhibition of endothelial cell proliferation, in vitro angiogenesis, and the down-regulation of cell adhesion–related genes by genistein combined with a cdna microarray analysis. Endothelium 13, 249–266. Pignatelli, P., Pulcinelli, F.M., Lenti, L., Gazzaniga, P.P., Violi, F., 1998. Hydrogen peroxide is involved in collagen-induced platelet activation. Blood 91 (2), 484–490. Pignatelli, P., Pulcinelli, F.M., Celestini, A., Lenti, L., Ghiselli, A., Gazzaniga, P.P., Violi, F., 2000. The flavonoids quercetin and catechin synergistically inhibit platelet function by antagonizing the intracellular production of hydrogen peroxide. Am. J. Clin. Nutr. 72 (5), 1150–1155. Pignatelli, P., Di Santo, S., Buchetti, B., Sanguigni, V., Brunelli, A., Violi, F., 2006. Polyphenols enhance platelet nitric oxide by inhibiting protein kinase C-dependent NADPH oxidase activation: effect on platelet recruitment. FASEB J. 20 (8), 1082–1089. Ras, R.T., Zock, P.L., Zebregs, Y.E., Johnston, N.R., Webb, D.J., Draijer, R., 2013. Effect of polyphenol-rich grape seed extract on ambulatory blood pressure in subjects with pre- and stage I hypertension. Br. J. Nutr. 110 (12), 2234–2241. Rendu, F., Brohard-Bohn, B., 2001. The platelet release reaction: granules' constituents, secretion and functions. Platelets 12, 261–273. Rezende, B.A., Cortes, S.F., Lemos, V.S., 2004. Mechanisms involved in the vasodilator effect of the flavanol floranol in rat small mesenteric arteries. Planta Med. 70, 465–467. Rezende, B.A., Cortes, S.F., De Sousa, F.B., Lula, I.S., Schmitt, M., Sinisterra, R.D., Lemos, V.S., 2009. Complexation with β-cyclodextrin confers oral activity on the flavonoid dioclein. Int. J. Pharm. 367, 133–139. Ribeiro-Santos, R., Carvalho-Costa, D., Cavaleiro, C., Costa, H.S., Albuquerque, T.G., Castilho, M.C., Ramos, F., Melo, N.R., Sanches-Silva, A., 2015. A novel insight on an ancient aromatic plant: the rosemary (Rosmarinus officinalis L.). Trends Food Sci. Technol. 45, 355–368. Rink, T.J., Sage, S.O., 1990. Calcium signaling in human platelets. Annu. Rev. Physiol. 52, 431–449. Ross, J.A., Kasum, C.M., 2002. Dietary flavonoids: bioavailability, metabolic effects, and safety. Ann. Rev. Nutr. 22, 19–34. Rull, G., Mohd-Zain, Z.N., Shiel, J., Lundberg, M.H., Collier, D.J., Johnston, A., Warner, T.D., Corder, R., 2015. Effects of high flavanol dark chocolate on cardiovascular function and platelet aggregation. Vasc. Pharmacol. 71, 70–78. Schmidt, E.M., Schmid, E., Münzer, P., Hermann, A., Eyrich, A.K., Russo, A., Walker, B., Gu, S., vom Hagen, J.M., Faggio, C., 2013. Chorein sensitivity of cytoskeletal organization and degranulation of platelets. FASEB J. 27, 2799–2806. Schumacher, E., Vigh, E., Molnár, V., Kenyeres, P., Fehér, G., Késmárky, G., Tóth, K., Garai, J., 2011. Thrombosis preventive potential of chicory coffee consumption: a clinical study. Phytother. Res. 25 (5), 744–748. Scull, C.M., Hays, W.D., Fischer, T.H., 2010. Macrophage pro-inflammatory cytokine secretion is enhanced following interaction with autologous platelets. J. Inflamm. 7, 53. Senchenkova, E., Seifert, H., Granger, D.N., 2015. Hypercoagulability and platelet abnormalities in inflammatory bowel disease. Semin. Thromb. Hemost. 41 (6), 582–589. Shattil, S., Brass, L., 1987. Induction of the fibrinogen receptor on human platelets by intracellular mediators. J. Biol. Chem. 262, 992–1000. Sheu, J.R., Hsiao, G., Chou, P.H., Shen, M.Y., Chou, D.S., 2004. Mechanisms involved in the antiplatelet activity of rutin, a glycoside of the flavonol quercetin, in human platelets. J. Agric. Food Chem. 52 (14), 4414–4418. Simons, A.L., Renouf, M., Hendrich, S., Murphy, P.A., 2005. Metabolism of glycitein (7,4′-dihydroxy-6-methoxy-isoflavone) by human gut microflora. J. Agric. Food Chem. 53 (22), 8519–8525. Stephen, J., Emerson, B., Fox, K.A., Dransfield, I., 2013. The uncoupling of monocyte– platelet interactions from the induction of proinflammatory signaling in monocytes. J. Immunol. 191, 5677–5683. Thon, J.N., Peters, C.G., Machlus, K.R., Aslam, R., Rowley, J., Macleod, H., Devine, M.T., Fuchs, T.A., Weyrich, A.S., Semple, J.W., 2012. T granules in human platelets function in TLR9 organization and signaling. J. Cell Biol. 198, 561–574.

Res. 54 (10), 1405–1413. Hao, H.Z., He, A.D., Wang, D.C., Yin, Z., Zhou, Y.J., Liu, G., Liang, M.L., Da, X.W., Yao, G.Q., Xie, W., Xiang, J.Z., Ming, Z.Y., 2015. Antiplatelet activity of loureirin A by attenuating Akt phosphorylation: In vitro studies. Eur. J. Pharmacol. 746, 63–69. Hathaway, D.R., Adelstein, R.S., 1979. Human platelet myosin light chain kinase requires the calcium-binding protein calmodulin for activity. Proc. Natl. Acad. Sci. 76, 1653–1657. Heijnen, H., van der Sluijs, P., 2015. Platelet secretory behaviour: as diverse as the granules … or not? J. Thromb. Haemost. 13 (12), 2141–2151. Heim, K.E., Tagliaferro, A.R., Bobilya, D.J., 2002. Flavonoid antioxidants: chemistry, metabolism and structure-activity relationships. J. Nutr. Biochem. 13, 572–584. Heiss, C., Kleinbongard, P., Dejam, A., Perré, S., Schroeter, H., Sies, H., Kelm, M., 2005. Acute consumption of flavanol-rich cocoa and the reversal of endothelial dysfunction in smokers. J. Am. Coll. Cardiol. 46, 1276–1283. Hollman, P.C., 2004. Absorption, bioavailability, and metabolism of flavonoids. Pharm. Biol. 42, 74–83. Hubbard, G., Stevens, J., Cicmil, M., Sage, T., Jordan, P., Williams, C., Lovegrove, J., Gibbins, J., 2003. Quercetin inhibits collagen‐stimulated platelet activation through inhibition of multiple components of the glycoprotein VI signaling pathway. J. Thromb. Haemost. 1, 1079–1088. Hung, C.C., Tsai, W.J., Kuo, L.M., Kuo, Y.H., 2005. Evaluation of caffeic acid amide analogues as anti-platelet aggregation and anti-oxidative agents. Bioorg. Med. Chem. 13 (5), (1791-1717). Jackson, S.P., 2007. The growing complexity of platelet aggregation. Blood 109, 5087–5095. Jeong, Y.J., Choi, Y.J., Kwon, H.M., Kang, S.W., Park, H.S., Lee, M., Kang, Y.H., 2005. Differential inhibition of oxidized LDL-induced apoptosis in human endothelial cells treated with different flavonoids. Br. J. Nutr. 93, 581–591. Karuppagounder, S., Madathil, S., Pandey, M., Haobam, R., Rajamma, U., Mohanakumar, K., 2013. Quercetin up-regulates mitochondrial complex-I activity to protect against programmed cell death in rotenone model of Parkinson's disease in rats. Neuroscience 236, 136–148. Kim, D.H., Jung, E.A., Sohng, I.S., Han, J.A., Kim, T.H., Han, M.J., 1998. Intestinal bacterial metabolism of flavonoids and its relation to some biological activities. Arch. Pharm. Res. 21 (1), 17–23. Kim, J.M., Yun-Choi, H.S., 2008. Anti-platelet effects of flavonoids and flavonoidglycosides from Sophora japonica. Arch. Pharm. Res. 31, 886–890. Kozlowska, A., Szostak-Wegierek, D., 2014. Flavonoids-food sources and health benefits. Rocz. Państwowego Zakładu Hig., 65. Kumar, S., Pandey, A.K., 2013. Chemistry and biological activities of flavonoids: an overview. Sci. World J. 2013, 162750. Lemos, V.S., Freitas, M.R., Muller, B., Lino, Y.D., Queiroga, C.E., Côrtes, S.F., 1999. Dioclein, a new nitric oxide-and endothelium-dependent vasodilator flavonoid. Eur. J. Pharmacol. 386, 41–46. Li, N., 2008. Platelet–lymphocyte cross-talk. J. Leukoc. Biol. 83, 1069–1078. Liang, M.L., Da, X.W., He, A.D., Yao, G.Q., Xie, W., Liu, G., Xiang, J.Z., Ming, Z.Y., 2015. Pentamethylquercetin (PMQ) reduces thrombus formation by inhibiting platelet function. Sci. Rep. 5, 11142. Lindahl, M., Tagesson, C., 1997. Flavonoids as phospholipase A2 inhibitors: importance of their structure for selective inhibition of group II phospholipase A2. Inflammation 21, 347–356. Ludwig, A., Lorenz, M., Grimbo, N., Steinle, F., Meiners, S., Bartsch, C., Stangl, K., Baumann, G., Stangl, V., 2004. The tea flavonoid epigallocatechin-3-gallate reduces cytokine-induced VCAM-1 expression and monocyte adhesion to endothelial cells. Biochem. Biophys. Res. Commun. 316, 659–665. Luzak, B., Kassassir, H., Rój, E., Stanczyk, L., Watala, C., Golanski, J., 2017. Xanthohumol from hop cones (Humulus lupulus L.) prevents ADP-induced platelet reactivity. Arch. Physiol. Biochem. 123 (1), 54–60. Maheswaraiah, A., Rao, L.J., Naidu, K.A., 2015. Anti-platelet activity of water dispersible curcuminoids in rat platelets. Phytother. Res. 29 (3), 450–458. Mantena, S.K., Baliga, M.S., Katiyar, S.K., 2006. Grape seed proanthocyanidins induce apoptosis and inhibit metastasis of highly metastatic breast carcinoma cells. Carcinogenesis 27, 1682–1691. Maynard, D., Heijnen, H., Horne, M., White, J., Gahl, W., 2007. Proteomic analysis of platelet α‐granules using mass spectrometry. J. Thromb. Haemost. 5, 1945–1955. Mazereeuw, G., Herrmann, N., Bennett, S.A., Swardfager, W., Xu, H., Valenzuela, N., Fai, S., Lanctôt, K.L., 2013. Platelet activating factors in depression and coronary artery disease: a potential biomarker related to inflammatory mechanisms and neurodegeneration. Neurosci. Biobehav. Rev. 37 (8), 1611–1621. Meng, S., Wu, B., Singh, R., Yin, T., Morrow, J.K., Zhang, S., Hu, M., 2012. SULT1A3mediated regiospecific 7-O-sulfation of flavonoids in Caco-2 cells can be explained by the relevant molecular docking studies. 9 (4), 862. Messina, F., Guglielmini, G., Curini, M., Orsini, S., Gresele, P., Marcotullio, M.C., 2015. Effect of substituted stilbenes on platelet function. Fitoterapia 105, 228–233. Mitchelmore, C., Troelsen, J.T., Spodsberg, N., Sjöström, H., Norén, O., 2000. Interaction between the homeodomain proteins Cdx2 and HNF1alpha mediates expression of the lactase-phlorizin hydrolase gene. Biochem. J. 346 (Pt 2), 529–535. Mladěnka, P., Zatloukalová, L., Filipský, T., Hrdina, R., 2010. Cardiovascular effects of flavonoids are not caused only by direct antioxidant activity. Free Radic. Biol. Med. 49, 963–975. Mower, R.L., Landolfi, R., Steiner, M., 1984. Inhibition in vitro of platelet aggregation and arachidonic acid metabolism by flavone. Biochem. Pharmacol. 33, 357–363. Muñoz, Y., Garrido, A., Valladares, L., 2009. Equol is more active than soy isoflavone itself to compete for binding to thromboxane A(2) receptor in human platelets. Thromb. Res. 123 (5), 740–744. Nabavi, S.F., Nabavi, S.M., Mirzaei, M., Moghaddam, A.H., 2012. Protective effect of

100

European Journal of Pharmacology 807 (2017) 91–101

C. Faggio et al.

distributed on external and internal membranes of resting platelets. Am. J. Pathol. 155, 2127–2134. Williamson, J., Cooper, R., Joseph, S., Thomas, A., 1985. Inositol trisphosphate and diacylglycerol as intracellular second messengers in liver. Am. J. Physiol. Cell Physiol. 248, C203–C216. Wright, B., Gibson, T., Spencer, J., Lovegrove, J.A., Gibbins, J.M., 2010a. Plateletmediated metabolism of the common dietary flavonoid, quercetin. PLoS One 5 (3), e9673. Wright, B., Moraes, L.A., Kemp, C.F., Mullen, W., Crozier, A., Lovegrove, J.A., Gibbins, J.M., 2010b. A structural basis for the inhibition of collagen‐stimulated platelet function by quercetin and structurally related flavonoids. Br. J. Pharmacol. 159, 1312–1325. Wright, B., Spencer, J.P., Lovegrove, J.A., Gibbins, J.M., 2013. Insights into dietary flavonoids as molecular templates for the design of anti-platelet drugs. Cardiovasc. Res. 97, 13–22. Wu, B., Kulkarni, K., Basu, S., Zhang, S., Hu, M., 2011. First-pass metabolism via UDPglucuronosyltransferase: a barrier to oral bioavailability of phenolics. J. Pharm. Sci. 100 (9), 3655–3681. Yang, Z., Zhu, W., Gao, S., Yin, T., Jiang, W., Hu, M., 2012. Breast cancer resistance protein (ABCG2) determines distribution of genistein phase II metabolites: reevaluation of the roles of ABCG2 in the disposition of genistein. Drug Metab. Dispos. 40 (10), 1883–1893. Yeaman, M.R., 2010. Platelets in defense against bacterial pathogens. Cell. Mol. Life Sci. 67, 525–544. Zern, T.L., Wood, R.J., Greene, C., West, K.L., Liu, Y., Aggarwal, D., Shachter, N.S., Fernandez, M.L., 2005. Grape polyphenols exert a cardioprotective effect in pre-and postmenopausal women by lowering plasma lipids and reducing oxidative stress. J. Nutr. 135, 1911–1917. Zubair, M.H., Zubair, M.H., Zubair, M.N., Zubair, M.M., Aftab, T., Asad, F., 2011. Augmentation of anti-platelet effects of aspirin. J. Pak. Med. Assoc. 61 (3), 304–307. Zufferey, A., Schvartz, D., Nolli, S., Reny, J.L., Sanchez, J.C., Fontana, P., 2014. Characterization of the platelet granule proteome: evidence of the presence of MHC1 in alpha-granules. J. Proteom. 101, 130–140.

Tian, X., Chang, L., Ma, G., Wang, T., Lv, M., Wang, Z., Chen, L., Wang, Y., Gao, X., Zhu, Y., 2016. Delineation of platelet activation pathway of scutellarein revealed its intracellular target as protein kinase C. Biol. Pharm. Bull. 39 (2), 181–191. Tran, H., Anand, S.S., 2004. Oral antiplatelet therapy in cerebrovascular disease, coronary artery disease, and peripheral arterial disease. JAMA 292, 1867–1874. Trischitta, F., Faggio, C., 2006. Effect of the flavonol quercetin on ion transport in the isolated intestine of the eel, Anguilla anguilla. Comp. Biochem. Physiol. C: Toxicol. Pharmacol. 143, 17–22. Tzeng, S.H., Ko, W.C., Ko, F.N., Teng, C.M., 1991. Inhibition of platelet aggregation by some flavonoids. Thromb. Res. 64, 91–100. Vieira-de-Abreu, A., Campbell, R.A., Weyrich, A.S., Zimmerman, G.A., 2012. Platelets: versatile effector cells in hemostasis, inflammation, and the immune continuum. Semin. Immunopathol. 34, 5–30. Walle, T., 2004. Absorption and metabolism of flavonoids. Free Radic. Biol. Med. 36, 829–837. Wang, S.B., Jang, J.Y., Chae, Y.H., Min, J.H., Baek, J.Y., Kim, M., Park, Y., Hwang, G.S., Ryu, J.S., Chang, T.S., 2015. Kaempferol suppresses collagen-induced platelet activation by inhibiting NADPH oxidase and protecting SHP-2 from oxidative inactivation. Free Radic. Biol. Med. 83, 41–53. Wang, W., Heideman, L., Chung, C.S., Pelling, J.C., Koehler, K.J., Birt, D.F., 2000. Cell‐ cycle arrest at G2/M and growth inhibition by apigenin in human colon carcinoma cell lines. Mol. Carcinog. 28, 102–110. Watson, S., Auger, J., McCarty, O., Pearce, A., 2005. GPVI and integrin αIIbβ3 signaling in platelets. J. Thromb. Haemost. 3, 1752–1762. Wei, Y., Wu, B., Jiang, W., Yin, T., Jia, X., Basu, S., Yang, G., Hu, M., 2013. Revolving door action of breast cancer resistance protein (BCRP) facilitates or controls the efflux of flavone glucuronides from UGT1A9-overexpressing HeLa cells. Mol. Pharm. 10 (5), 1736–1750. Weseler, A.R., Ruijters, E.J., Drittij-Reijnders, M.J., Reesink, K.D., Haenen, G.R., Bast, A., 2011. Pleiotropic benefit of monomeric and oligomeric flavanols on vascular health–a randomized controlled clinical pilot study. PLoS One 6 (12), e28460. White, J.G., Clawson, C., 1980. The surface-connected canalicular system of blood platelets–a fenestrated membrane system. Am. J. Pathol. 101, 353–364. White, J.G., Krumwiede, M.D., Escolar, G., 1999. Glycoprotein Ib is homogeneously

101