Anthocyanins: from sources and bioavailability to cardiovascular

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Anthocyanins: from sources and bioavailability to cardiovascular health benefits and molecular mechanisms of action Irena Krga, and Dragan Milenkovic J. Agric. Food Chem., Just Accepted Manuscript • DOI: 10.1021/acs.jafc.8b06737 • Publication Date (Web): 30 Jan 2019 Downloaded from http://pubs.acs.org on January 31, 2019

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Journal of Agricultural and Food Chemistry

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Anthocyanins: from sources and bioavailability to cardiovascular health benefits

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and molecular mechanisms of action

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Irena Krga 1,2 and Dragan Milenkovic 2,3*

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1 Centre

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University of Belgrade, Belgrade, Serbia;

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2 Université

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France;

of Research Excellence in Nutrition and Metabolism, Institute for Medical Research,

Clermont Auvergne, INRA, UNH, CRNH Auvergne, F-63000 Clermont-Ferrand,

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3 Department

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University of California Davis, Davis, California, 95616, United States of America

of Internal Medicine, Division of Cardiovascular Medicine, School of Medicine,

12 13

* Corresponding author:

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E-mail address: [email protected] (D. Milenkovic).

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Phone: +33(0)4 73 62 45 79

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Fax: +33(0)4 73 62 46 38

1 ACS Paragon Plus Environment


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Abstract

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Anthocyanins are phytochemicals widely found in plant foods, with berries and fruit-derived

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beverages as main dietary sources. Accumulating evidence suggests the positive role of

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anthocyanins in preserving cardiovascular health. Epidemiological data show an association

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between anthocyanin intake and lower risk of myocardial infarction and cardiovascular

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disease-related mortality. Clinical studies report the beneficial effects of the consumption of

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different anthocyanin-rich sources on surrogate markers of cardiovascular risk. Animal and in

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vitro evidence suggest the protective role of anthocyanins in dysfunctions related to the

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development of cardiovascular diseases. Still, the underlying molecular mechanisms of

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anthocyanin action seem complex and are not entirely clear. This review aims to give a

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comprehensive update on anthocyanins and their cardioprotective properties. It provides

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information on their sources, quantities consumed through diet, absorption, bioavailability,

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cardiovascular properties and underlying mechanisms of action including their effects on gene

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and protein expression, interaction with cell signaling pathways and miRNAs.

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Keywords: anthocyanin; sources; bioavailability; cardioprotection; mechanisms of action


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1. Introduction

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Cardiovascular diseases (CVD) are a group of disorders that affect heart and blood vessels and

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represent the leading cause of morbidity and mortality worldwide 1. Diet plays an essential role

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in the initiation and progression of CVD but also presents a significant lifestyle factor for the

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prevention of these and other diseases. Data from epidemiological and clinical studies have

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shown an inverse association between CVD development and consumption of diets rich in

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fruits and vegetables 2,3. The health properties of fruits and vegetables can be ascribed not only

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to their low-caloric value but also to their high content in fiber, essential micronutrients and

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bioactive compounds like phytochemicals. With over 500 species found in the human diet,

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polyphenols represent the largest and most prevalent of phytochemicals 4. In addition to being

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present in fruits and vegetables, they are also found in plant-derived beverages such as wine,

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fruit juices, herbal tea, and coffee, contributing to a dietary intake of more than 1 g/day 5.

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A growing body of evidence suggests that polyphenols contribute to the cardiovascular health

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benefits associated with diets rich in fruits and vegetables

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anti-inflammatory, antioxidant and antithrombotic activates by acting through complex

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mechanisms of action. Polyphenols present the capacity to interact with the cell membranes

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leading to changes in their structure and physical characteristics that could disturb cell function

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9.

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transcription factors and affect the expression of genes 7,10.

2,3,6–8.

These compounds can exert

Also, they can interact with cellular receptors, modulate the activities of enzymes and

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Over the past two decades the interest in anthocyanins, polyphenols particularly abundant

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in berries and berry-derived products, has dramatically increased, especially due to the

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cardioprotective effects associated with the consumption of anthocyanin-rich plant foods. This

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review aimed to give a complete update on anthocyanins and their cardiovascular health

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properties. It provides information on their sources, quantities consumed through normal diets,

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their absorption and bioavailability, cardiovascular effects as well as underlying mechanisms 3 ACS Paragon Plus Environment


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of their action by considering their impact on the expression of genes, proteins, interaction with

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cell signaling pathways and miRNA.

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2. Anthocyanins: chemistry, consumption and bioavailability

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2.1. Chemical structure and characteristics of anthocyanins

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Anthocyanins are water-soluble plant pigments that give red, purple and blue coloration of

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many fruits, flowers, and leaves. They are glycosylated, polyhydroxy, or polymetoxy

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derivatives of flavylium cation (2-phenylchromenylium)

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anthocyanins and 27 anthocyanidins (the sugar-free, aglycone forms of anthocyanins)

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identified in nature. However, only six anthocyanidins: cyanidin, delphinidin, pelargonidin,

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peonidin, malvidin, and petunidin are widely distributed in the human diet, accounting for more

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than 90% of all known anthocyanins 11 (Figure 1). The diversity of anthocyanin structure comes

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from differences in the 1) number and location of hydroxyl groups and a degree of their

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methylation, 2) nature, number and position of sugar attached to aglycone and 3) nature and

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number of aliphatic and aromatic acids linked to these sugars. Glucose, galactose, rutinose,

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rhamnose, arabinose, and xylose are commonly attached to anthocyanidins as mono-, di-, or

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trisaccharides mainly at C3-position of the C-ring or C5 or C7-position of the A-ring. These

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sugars can also be acylated with aromatic acids such as p-coumaric, ferulic, caffeic and p-

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hydroxybenzoic acids and aliphatic acids like malonic, acetic, malic and oxalic acids 11,12.

11.

There are 702 different

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Anthocyanins are reactive and very unstable compounds. Their stability can be affected by

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pH, temperature, light as well as the presence of oxygen, enzymes, other flavonoids, proteins

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and metal ions

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which relative abundance at equilibrium varies with pH and anthocyanidin structure.

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Anthocyanins are the most stable at low pH (1-3) where they occur as flavylium cations that

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are red-colored. As the pH increases, flavylium cation transforms either to the blue quinoidal

12.

In aqueous solution, anthocyanins can form several structural isoforms,


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base (pH 2-4) or to colorless hemiketal that further undergoes ring opening and produces the

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pale yellow chalcone (pH 5-6) 11. Chemical degradation of chalcone can further give rise to

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phenolic acids 11,12.

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2.2. Dietary consumption of anthocyanins

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Anthocyanins are substantial components of the human diet. They are regularly consumed in

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many fruits and fruit-derived products (e.g., wines, juices, and jams) and some dark-colored

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vegetables and cereals (e.g., eggplant, red onion, red cabbage, and black rice). They are also

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present in the diet as the coloring agent E163 that is increasingly used in the food industry for

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various foods and beverages. Among fruits, the most commonly eaten anthocyanin sources are

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berry fruits belonging to Vitaceae family (species Vitis vinifera), Rosaceae family (e.g.,

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strawberries, raspberries, blackberries, cherries, apples, plums) and genera Ribes (e.g.,

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blackcurrants) and Vaccinium (e.g., bilberries, blueberries, cranberries) of Ericaceae family 11.

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In these dietary sources anthocyanin concentration can reach several hundreds of mg in 100 g

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of fresh weight (F.W.), providing substantial anthocyanin doses in a single serving. The highest

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anthocyanin concentrations are found in elderberries (664-1816 mg/100 g F.W.), chokeberries

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(410-1480 mg/100 g F.W.), bilberries (300-698 mg/100 g F.W.), raspberries (20-687 mg/100

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g F.W.), blackcurrants (130-476 mg/100 g F.W.), blackberries (82.5-325.9 mg/100 g F.W.) and

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blueberries (61.8-299.6 mg/100 g F.W.) 13. It is noteworthy that anthocyanin concentrations in

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foods are influenced by genetic, environmental and agronomic factors such as light,

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temperature, humidity, fertilization, food processing and storage conditions 13. For example,

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when comparing different forms of processed blueberries, anthocyanins are better preserved in

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canned fruits (around 70%) and puree (57%) than in clarified juice (31%)

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produced blueberry juices have higher anthocyanin levels than those stored for six months at

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4°C (around 10% loss) and 25 °C (about 50% loss) 15.

14.

Also, freshly



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Daily anthocyanin intake varies greatly depending on the dietary habits of the studied

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population, which are influenced by socioeconomic, demographic and lifestyle factors. They

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are still poorly established, mainly due to the absence of available information in food

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composition databases and variations in results depending on the used dietary assessment. The

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estimated mean intakes in Europe range from 64.9 mg/day (Italy) to 19.8 (Netherlands) for

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men, and from 44.1 mg/day (Italy) to 18.4 (Spain) for women 16. In the USA, the reported mean

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consumption is 12.6 mg/day for women and 10.5 mg/day for men 17. Fruits, especially grapes,

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apples, pears and berries are responsible for approximately 50% of estimated habitual intake

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in Europe, while wines contribute by around 21%

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anthocyanin intakes are berries (20%), wines (16%), grapes (11%) and bananas (11 %) 17.

16.

In the USA the main contributors to

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2.3. Anthocyanin bioavailability

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Ingested dietary anthocyanins need to be available in the circulation and tissues to exert the

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effect in the human body. Bioavailability represents a portion of the ingested dose of a

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compound that reaches the general circulation and specific sites where it can exert its action 12.

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Anthocyanin bioavailability has been previously reported to be very low, with the recovery of

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less than 1% of the ingested anthocyanin dose 18. However, recent human bioavailability study

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that used isotopically labeled anthocyanin cyanidin-3-glucoside has reported the extensive

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anthocyanin metabolism and a recovery of 12.4% 19. These data suggest that the anthocyanin

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bioavailability may be much higher than previously thought due to newly identified

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metabolites. Therefore, a good understanding of the fate of dietary anthocyanins in the human

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body following their intake, i.e., their absorption, distribution, metabolism and excretion

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(ADME) is of great importance for assessing their possible biological effects.

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2.3.1. Anthocyanin absorption and metabolism

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In foods, anthocyanins are present as glycosides. After ingestion, these compounds move

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through different regions of the gastrointestinal tract (GIT) (Figure 2) where the specific pH,

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composition of the dissolved gases and metabolic activity can lead to their degradation and

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metabolism

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anthocyanin stability, allowing them to persist in glycoside form. The presence of glycosides

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in the human circulation within minutes of consumption suggests that unlike other flavonoids

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the ingested anthocyanins can be absorbed intact from the stomach

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are hydrophilic molecules that cannot pass the cell membranes by passive diffusion a

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transporter system needs to be involved. An organic anion membrane carrier named

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bilitranslocase that is expressed in the gastric mucosa as well as in the liver, kidneys and

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vascular endothelium has been proposed to mediate anthocyanin transport

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inhibition of its transport activity by quinoidal forms of different anthocyanins in vitro suggests

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that this transporter could be responsible for anthocyanins quick transport into the portal and

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general circulations 21. The involvement of glucose transporter (GLUT) 1 in the transport of

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anthocyanin glucosides has also been suggested 22. However, these proposed mechanisms of

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anthocyanin gastric absorption are based only on findings from in vitro studies, and their

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relevance to the absorption and metabolism in humans is still uncertain. The use of in vivo

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models, such as the genetic knockout mice, could elucidate the anthocyanin gastric absorption

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via the proposed transporters. Nevertheless, caution needs to be taken when using gene

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knockout model since deletion of one carrier can sometimes cause the alteration in the

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expression of other transporters/enzymes and change the physiology of the animal 23.

18.

In the stomach, the low pH (1.5-4) provides favorable conditions for

12,20.

Since anthocyanins

12,21.

Competitive

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The main site of anthocyanin absorption is the small intestine. Similar to other flavonoids,

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anthocyanins can there undergo deglycosylation (i.e., cleavage of the glycoside) producing

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lipophilic aglycones, which can then enter the epithelial cells by passive transport. 7 ACS Paragon Plus Environment


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Deglycosylation can be mediated by β-glucosidase in the intestinal lumen and lactase-

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phloridzin hydrolase in the brush border of the intestinal epithelial cells. Alternatively,

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absorption could involve the active transport of intact glycosides into the epithelial cells by the

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sodium-dependent glucose transporter 1 (SGLT1) or GLUT2 and their subsequent

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deglycosylation by cytosolic β-glucosidase 18,20. However, due to some contradictory results,

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the involvement of SGLT1 in the anthocyanin absorption is still uncertain 20.

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Anthocyanin aglycones that enter the intestinal epithelial cells may be metabolized there

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before reaching portal circulation. They undergo metabolic detoxification typical for many

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xenobiotics, which increases their hydrophilicity and facilitates elimination from the body

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through bile and urine. This metabolism includes the phase I (oxidation, reduction, hydrolysis

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reactions) and phase II metabolism (conjugation reaction). In the intestine, anthocyanins can

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undergo methylation, sulfation, and glucuronidation, mediated by phase II metabolizing

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enzymes

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glucuronosyltransferase, respectively 18,20. These reactions can also take place in the liver (i.e.,

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the most important site of xenobiotic metabolism) and kidneys. Consequently, following the

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intake of anthocyanin-rich foods, the methylated, sulfated and glucuronidated anthocyanins are

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detected in human plasma and urine 12,18,20.

catechol-O-methyltransferase,

sulfotransferase

and

uridine-5’-diphospho-

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Before conjugation, anthocyanin aglycones can alternatively undergo degradation to

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phenolic acids and aldehydes within the intestinal lumen or epithelial cells. Anthocyanin

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degradation can also be a result of the activity of colonic microbiota. Anthocyanins that reach

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the colon are exposed to 300-500 different bacterial species, with Bifidobacterium,

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Bacteroides, Eubacterium, and Clostridium presenting the most abundant genera. Gut

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microbiota releases many deglycosylation enzymes that cleave the sugar moiety, giving rise to

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aglycones that further undergo ring opening to produce different phenolic acids (e.g.,

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protocatechuic acid (PCA), vanillic, syringic, ferulic, hippuric acids) or aldehydes

20,24.


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Consequently, the portion of ingested anthocyanin forms decreases along the GIT, whereas the

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portion of phenolic acids increases. The products of anthocyanin degradation may also be

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absorbed from the intestines probably through epithelial monocarboxylic acids transporters and

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further metabolized in the liver or kidneys 24,25. Anthocyanins that are not absorbed in the GIT

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are eliminated from the body through feces.

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2.3.2. Anthocyanins in the circulation

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The results from the available human bioavailability studies with anthocyanin-rich foods show

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that anthocyanins and their phase II conjugates appear rapidly in the circulation. They reach

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the maximal concentration of around 100 nM within 1.5 h and disappear from the bloodstream

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by 6 h post-consumption (Supporting Information, Table S1). The maximal plasma

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concentrations (Cmax) of total anthocyanins reported in these studies were in the range of 1.4-

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591.7 nM (Table S1), while Cmax of individual anthocyanins in human plasma varied from

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0.06 to 367 nM (Figure 3). It should be noted that the plasma anthocyanin levels in most of

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these bioavailability studies were proportional to their levels in tested food. Therefore, the

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concentrations of the individual glycosides reached in circulation after anthocyanin intake

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could hypothetically be much higher depending upon intake.

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In addition to anthocyanins and their phase II conjugates, several anthocyanin bioavailability

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studies have also reported the presence of other compounds in plasma (e.g., different phenolic

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acids) that might present products of anthocyanin metabolism. However, since their origin was

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hard to trace, these compounds were ascribed to anthocyanin food source used in studies. A

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study providing 500 mg of isotopically labeled cyanidin-3-glucoside to eight male subjects also

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detected different phenolic acids (e.g., PCA, vanillic, ferulic, hippuric acids) and their phase I

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and phase II metabolites in the circulation 19,25. The use of isotope-labeling on both A- and B-

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rings of this anthocyanin allowed authors to unequivocally define the origin of identified 9 ACS Paragon Plus Environment


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phenolic compounds and report them as anthocyanin-derived metabolites. Interestingly, unlike

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their parent anthocyanin that reached Cmax of 141 nM by 1.8 h and disappeared from the

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bloodstream by 6 h, metabolites peaked (0.2-2 M) at around 10 h and were detectable in the

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circulation up to 48 h post-ingestion. Furthermore, metabolites displayed biphasic serum

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profiles, with the first peak between 0 and 5 h and a second, more pronounced peak between 6

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and 48 h that corresponded to absorption of anthocyanin bacterial metabolites from the colon.

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Therefore, this study provided evidence of the extensive anthocyanin metabolism in the human

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body. Furthermore, it demonstrated that metabolites, especially those derived from microbial

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metabolism, are the dominant anthocyanin forms in the circulation and could potentially

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contribute to the beneficial health effects associated with the consumption of anthocyanin-rich

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sources.

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3. Cardiovascular health benefits of anthocyanins

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3.1. Evidence from epidemiological studies

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Accumulating evidence suggests the protective effects of anthocyanin consumption against

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CVD. Data from several epidemiological studies have reported an inverse correlation between

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anthocyanin intake and risk of CVD or CVD-related mortality. In a prospective study,

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following 34489 healthy postmenopausal women (55-69 years old) for 16 years, the

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anthocyanin intake was associated with a 12% and 9% lower risk of coronary heart disease

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(CHD) and CVD mortality, respectively 26. Similarly, a study of 38180 men and 60289 women

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(mean age of 70 and 69 years) with 7-year follow-up, observed a 21% reduction in risk of CHD

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mortality and 14% in CVD mortality in men and women when comparing higher (≥ 16.7

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mg/day) with lower ( 5.5 mg/day) anthocyanin intakes 27. Higher habitual anthocyanin intakes

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were also inversely associated with a risk of total myocardial infarction (MI) in premenopausal

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women 28 and non-fatal MI in men 29. The magnitude of this effect of anthocyanin consumption 10 ACS Paragon Plus Environment

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was higher in the study following 93600 premenopausal women (25-42 years old) for 18 years,

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with the observed 32% decrease in MI risk when comparing extremes of anthocyanin

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consumption (2.5 and 25.1 mg/day) 28. Additionally, a food-based analysis in this study showed

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that the combined intake of strawberries and blueberries tended to be associated with a reduced

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risk of MI. A 34% lower risk of MI was observed in women that ate more than three portions

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per week compared with those that rarely consumed these anthocyanin-rich fruits 28.

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Several cross-sectional and prospective studies have also shown the inverse association

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between anthocyanin intake and biomarkers of CVD risk, providing possible mechanistic

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support for the reported reductions in risk of CHD or CVD-related mortality. In a cross-

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sectional study investigating 1898 women aged between 18 and 75 years, higher anthocyanin

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intake was associated with significantly lower central blood pressure, mean arterial pressure

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and pulse wave velocity (PWV), direct measures of atherosclerosis and arterial stiffness 30. In

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this study, a 44-mg increase in anthocyanin intake was associated with a reduction in PWV by

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3.9% and a decrease in systolic blood pressure of 3 mmHg, suggesting that these effects could

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be easily achieved by incorporating around 1 to 2 portions of berries in an everyday diet. In

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several cross-sectional studies, higher anthocyanin intakes were also inversely correlated with

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different inflammatory biomarkers (e.g., IL-18 and C-reactive protein (CRP)) and the overall

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inflammation score that combined several cytokines, markers of acute inflammation and

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oxidative stress 31–33. Furthermore, a large prospective study following 156957 subjects for 14

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years reported that the higher anthocyanin intake, mainly from strawberry and blueberry

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consumption, is associated with an 8% reduction in risk of hypertension 6. This reduction was

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the most pronounced in premenopausal women, which correlated with a decrease in risk of

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MI 28.



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Results from the available epidemiological studies are promising regarding the role of

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anthocyanins in reducing the risk factors for CVD, reporting the associations that may have

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important public health implications.

258 259

3.2. Evidence from clinical studies

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As a result of these epidemiological studies, numerous randomized controlled trials (RTCs)

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have been initiated aiming to provide cause-effect relationships that could explain the role of

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anthocyanins in CVD prevention. Results of several meta-analyses of RCTs investigating the

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effects of the consumption of different anthocyanin-rich sources on surrogate markers of CVD

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risk, such as hypertension, lipid profiles, and endothelial dysfunction support the

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cardiovascular health benefits of anthocyanins

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reported that the consumption of berries and purified anthocyanins (2.2–1230 mg

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anthocyanins/day) significantly increases HDL-cholesterol and reduces LDL-cholesterol, TG,

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systolic blood pressure, diastolic blood pressure as well as the inflammatory markers CRP and

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TNFα

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subjects with age greater or equal to 50 years and those with the increased risk of CVD are

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more susceptible to the protective effects of anthocyanin consumption. Results of another meta-

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analysis of 99 RCTs showed that the consumption of anthocyanin-containing products, such as

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berries, red grapes and wines significantly lowers both systolic and diastolic blood pressure

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independently of the participants’ health status

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found to reduce total cholesterol levels and increase flow-mediated vasodilatation (FMD) 35.

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The improvements in FMD were also reported by Fairlie-Jones and colleagues, in a meta-

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analysis of 24 RCTs that examined the effects of anthocyanin-rich sources (foods, extracts or

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purified anthocyanins) on measures of vascular reactivity and stiffness

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analysis showed significant increases in FMD following both acute (1-8 h) and chronic (up to

34.

34–36.

The recent meta-analysis of 45 RCTs

The subgroup analysis in this study further revealed that overweight individuals,

35.

Additionally, berry supplementation was

36.

Results from this


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6 months) supplementations with anthocyanins (1 to 724 mg/day). Furthermore, a significant

281

improvement in PWV was observed after acute supplementation with anthocyanins.

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Unlike the relatively large number of RCTs investigating the effect of consumption of

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anthocyanin-rich sources on lipid profiles, blood pressure or vascular function, the number of

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studies assessing the impact on platelets is limited. Platelets contribute to the development of

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CVD by their interaction with 1eukocytes, endothelial cells, and other platelets that occur upon

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platelet activation. The increased platelet activation and formation of platelet-leukocyte

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aggregates are the biomarkers of acute CVD events and are more and more regarded as the

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marker of CVD risk 37. Despite the restricted number of studies, the available data do suggest

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that 4- to 24-week-long dietary interventions with anthocyanin-rich fruits or purified

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anthocyanins (202-320 mg anthocyanins/day) can affect platelet function by decreasing

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secretion of platelet chemokines, lowering platelet activation and their aggregation with

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leukocytes, and reducing thrombogenesis 38–41.

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It is important to note that together with a high number of RTCs that reported beneficial

294

effects of anthocyanin consumption on different surrogate markers of CVD risk there are

295

studies that failed to do so 42–45. These discrepancies in results may be due to differences in the

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studied population, baseline characteristics of subjects, duration of the interventions and the

297

anthocyanin concentrations in the tested anthocyanin-containing products. A high

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interindividual variation in the metabolism of plant-food bioactives, however, seems to be the

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critical factor contributing to differential responsiveness observed in RCTs. The

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polymorphisms of genes encoding enzymes involved in phase I and II metabolism may

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contribute to this variability 46. Nevertheless, microbiota is thought to be a main factor due to

302

its essential contribution to the metabolism of anthocyanins

303

show the high interindividual variability in type and levels of microbial metabolites created

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following the ingestion of anthocyanin-rich sources

25,47.

24,25.

Results from a few studies

These differences in the production 13

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of metabolites depend on the composition and function of microbiota that colonizes the gut of

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each individual and could result in different biological effects. The interactions between

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anthocyanins and microbiota seem to be bidirectional 48. In a recent RCT, high levels of fecal

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Bifidobacteria have been associated with increased levels of anthocyanin microbial metabolites

309

after the consumption of red wine 49. Additionally, a RCT investigating the effects of moderate

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consumption of red wine on modulation of the gut microbiota composition in metabolic

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syndrome patients revealed significant increases in fecal Bifidobacteria, Lactobacillus

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(intestinal barrier protectors) and butyrate-producing bacteria (Faecalibacterium prausnitzii

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and Roseburia) as well as the reduction in the levels of less desirable groups of bacteria

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(Escherichia coli and Enterobacter cloacae)

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microbiota composition were associated with the observed improvements in blood pressure

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and blood glucose levels in these subjects. Therefore, it seems that gut microbiota not only

317

plays a substantial role in anthocyanin health effects by producing various potentially bioactive

318

metabolites but also anthocyanin consumption can modulate the composition and activity of

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microbiota, thus enhancing its potential health effects.

50.

The red wine-induced modulations in gut

320

In conclusion, evidence from clinical studies shows that the intake of anthocyanin-rich

321

sources can improve different markers of CVD risk. However, well-designed RCTs that focus

322

on the effects of pure compounds, interindividual variability in anthocyanin metabolism and

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the importance of the microbiota are needed to strengthen the available evidence and establish

324

the role of anthocyanins in the prevention of CVD. Additionally, further research is required

325

to ascertain the impact of anthocyanin consumption on changes in microbiota and associated

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health benefits.

327 328

3.3. Evidence from animal studies


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Findings from animal studies also support the beneficial effects of anthocyanins on

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cardiovascular health. Majority of these data come from studies that used apolipoprotein E

331

knockout (ApoE−/−) mice that spontaneously develop atherosclerosis. Atherosclerosis

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represents a primary underlying pathologic process responsible for CVD development. This

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chronic inflammatory disorder of large and medium-sized arteries is characterized by

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atherosclerotic lesions that consist of accumulations of lipids, cellular and fibrous elements that

335

disturb the blood flow and can lead to MI and stroke 1. Several nutritional interventions have

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reported that supplementation of diet with anthocyanin-rich extracts or pure compounds

337

(0.003-1%) reduce the formation of atherosclerotic lesions in the aorta of ApoE−/− mice 51–53.

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For example, we have observed an average decrease of 25% in the aortic sinus lesion area of

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ApoE−/− after 20-week supplementation with a diet containing 0.02% bilberry extract (around

340

0.01% anthocyanins) 51. This supplementation may correspond to the equivalent human intake

341

of around 30 mg of anthocyanidins per day when expressed through diet content (for a human

342

food intake estimate of 500 g of dry weight). Anthocyanin supplementation was also shown to increase HDL 54, lower blood pressure 55–

343 344

57,

345

endothelium-dependent vasorelaxation

346

development 59,60 and increase survival following induced MI 61 in different animal models of

347

CVD.

LDL

57,58,

total cholesterol, TG

54,55,58

and inflammatory markers 56,

55,57,

as well as improve

decrease platelet hyperactivity, thrombus

348 349

3.4. Molecular mechanisms of action of anthocyanins underlying their cardiovascular health

350

properties

351

Numerous in vitro and in vivo studies have been performed to identify mechanisms by which

352

anthocyanins exert their cardioprotective effects. Many studies suggested that anthocyanin

353

action is mostly localized at the endothelial level, contributing to vascular homeostasis. 15 ACS Paragon Plus Environment


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354

Impaired endothelial function drives the development of atherosclerosis and is associated with

355

cardiovascular risk factors (e.g., hypertension, dyslipidemia) and characterized by reduced

356

endothelial vasodilatation, prothrombotic and proinflammatory properties of the endothelium

357

62.

358

leukocytes and their subsequent transendothelial migration into the blood vessel wall. In the

359

subendothelium, leukocytes differentiate into activated macrophages, take up oxidized

360

lipoproteins and form foam cells, which accumulation eventually leads to the formation of

361

atherosclerotic lesions and CVD development 1.

These changes in endothelial function promote the recruitment and adhesion of circulating

362

The mechanisms underlying the anthocyanin biological effects were previously ascribed to

363

their direct antioxidant properties, i.e., ability to transfer hydrogen (electron) to reactive oxygen

364

species and neutralize them. The imbalance between the production of reactive oxygen species

365

and antioxidant defenses in favor of the former that leads to disruption of redox signaling and

366

molecular damage (termed oxidative stress) is an important promoter of inflammatory reactions

367

and one of the hallmarks of endothelial dysfunction and atherosclerosis 62. It was proposed that

368

by exerting the direct antioxidant effect against reactive oxygen species, anthocyanins could

369

prevent LDL oxidation and associated inflammatory responses, thus attenuating the

370

development and progression of atherosclerosis 63. However, more recent studies have revealed

371

the implication of more complex molecular mechanisms of action including modulation of

372

gene expression, cell signaling and miRNA expression.

373 374

3.4.1. Effect on gene expression

375

Rather than acting as direct antioxidants, several in vitro studies reported that anthocyanins and

376

their microbial metabolites could modulate the expressions of genes coding for both anti- and

377

pro-oxidant enzymes. Anthocyanins at concentrations ranging from 1 µM to 40 µM have been

378

shown to increase the expressions of genes encoding enzymes involved in antioxidant defenses 16 ACS Paragon Plus Environment

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379

such as heme oxygenase-1 (HO-1) and NADPH quinone oxidoreductase 1 (NQO1) under basal

380

conditions or following the exposure of endothelial cells to different inflammatory mediators

381

64–67.

382

concentrations significantly increased the expressions of genes encoding HO-1, NQO1,

383

glutamate-cysteine ligase catalytic subunit (GCLC) and glutamate-cysteine ligase regulatory

384

subunit (GCLM) in gamma-radiated endothelial cells

385

HO-1 and GCLM were recently reported in hydrogen peroxide-treated endothelial cells pre-

386

exposed for 18 h to the mixture of phenolic acids observed in human plasma following the

387

intake of blueberry juice (PCA, 2-hydroxyhippuric, 4-hydroxyhippuric, syringic, gentisic,

388

vanillic, trans-ferulic, p-coumaric, dihydroferulic, dihydrocaffeic, dihydro-m-coumaric, and

389

homovanillic acids) 69. Few studies have also shown that anthocyanins and their metabolites

390

(at 0.1-200 µM levels) can downregulate the expressions of genes coding for different subunits

391

of NADPH oxidase, a pro-oxidant enzyme that represents a significant source of oxygen radical

392

production 70–72.

393

Similar to in vitro evidence, reduced expressions of genes encoding NADPH oxidase subunits,

394

NOX2 and NOX4, were observed in the coronary artery endothelial cells isolated from db-/db-

395

mice following a 10-week supplementation with strawberries 73. Furthermore, genes encoding

396

antioxidant enzymes like glutathione reductase, thioredoxin reductase 1 and superoxide

397

dismutase 1 and 2 were upregulated in the aorta of ApoE−/− mice after 20 weeks of dietary

398

supplementation with blueberries 53. This nutrigenomic effect was accompanied by the reduced

399

formation of atherosclerotic lesions in the aorta of these mice, suggesting that the modulation

400

of expression of genes involved in regulation of redox balance could present one of the

401

mechanisms by which anthocyanins and their metabolites could achieve their cardioprotective

402

effects. However, based on this evidence we cannot conclude that these compounds could

Ma et al. showed that the pre-treatment of endothelial cells with ferulic acid at 0.1-10 µM

68.

Similarly, increased expressions of



Page 18 of 46

403

restore the redox balance, and this indirect antioxidant action most probably contribute to other

404

mechanism mediating the observed effects.

405

Anthocyanins have also been shown to increase the expression of NOS3 and decrease EDN1

406

expression in endothelial cells 74,75[169,175,176]. NOS3 encodes nitric oxide (NO) synthase 3,

407

an enzyme that produces the vasoactive NO from L-arginine, leading to the NO-dependent

408

relaxation of vascular smooth muscle. EDN1 gene codes for a preproprotein that is

409

proteolytically processed to generate an endothelium-derived vasoactive peptide endothelin 1

410

that leads to the constriction of vascular smooth muscle. Decreased production of endothelial-

411

derived NO, the upregulation of EDN1 and subsequently increased contractibility are important

412

characteristics of endothelial dysfunction and pathogenesis of CVD 7. Therefore, these limited

413

studies suggest that the increased NOS3 expression and NO synthesis, as well as the

414

downregulation of EDN1, could represent the molecular mechanisms by which anthocyanins

415

contribute to the improved endothelial function reported in RTCs. However, it should be noted

416

that the main constraint of these studies is the use of high concentrations of anthocyanins (25-

417

100 µM) that are far from achievable in circulation. Also, the evidence of the impact of

418

anthocyanin metabolites on the expression of these genes in endothelial cells is lacking.

419

Majority of the available mechanistic evidence show that anthocyanins and their metabolites

420

can modulate the expression of genes regulating the inflammatory responses. During the

421

inflammatory insult, endothelial cells produce the proinflammatory cytokines like TNF and

422

interleukins that stimulate the surface expression of different cell adhesion molecules such as

423

the vascular cell adhesion molecule 1 (VCAM-1), intercellular adhesion molecule 1 (ICAM-1)

424

and E-selectin. Endothelium will also produce chemotactic cytokines such as monocyte

425

chemoattractant protein-1 (MCP-1) or IL-8 that guide leukocytes into the subendothelial space

426

7.

427

M have been reported to modulate the expressions of genes encoding the adhesion molecules

Anthocyanin-based extracts and pure compounds at concentrations ranging from 0.1 to 100


Page 19 of 46


428

such as VCAM-1, ICAM-1 and E selectin, cytokines like IL-6, IL-8 and MCP-1 as well as the

429

transcription factor NF-B in endothelial cells exposed to different inflammatory mediators 76–

430

81.

431

79,82–84.

432

ICAM-1 and IL-8 in palmitate-activated endothelial cells pre-treated with the mixture of

433

metabolites (hydroxyhippuric acid, hippuric acid, benzoic acid-4-sulfate, isovanillic acid-3-

434

sulfate, and vanillic acid-4-sulfate) detected in the human plasma following blueberry

435

consumption

436

stimulated endothelial cells pre-treated with the mixtures of metabolites identified in human

437

circulation 1, 6 and 24 h after the ingestion of 500 mg isotopically labeled cyanidin-3-glucoside

438

83.

439

human gut microorganisms (Enterococcus faecalis, Hafnia alvei) were recently reported to

440

lower the expressions of genes encoding E-selectin, ICAM-1, VCAM-1, IL-8, IL-6 and NF-

441

B-p65 in TNF-activated endothelial cells co-cultured with Caco-2 cells 84. In our latest work,

442

using a macroarray analysis, we showed that physiologically relevant mixtures of anthocyanins

443

and their microbial metabolites modulated the expressions of different genes involved in

444

chemokine signaling, cell adhesion and migration in TNFα-stimulated endothelial cells 10. In

445

the majority of these studies, the reported nutrigenomic effects were accompanied by a

446

decrease in monocyte adhesion to activated endothelial cells

447

evidence, the inhibition of aortic expression of several proinflammatory factors and adhesion

448

molecules involved in the recruitment of inflammatory cells (e.g., VCAM-1, ICAM-1 and

449

TNF) were also observed in studies with ApoE−/− mice that consumed diet enriched in

450

anthocyanin-based extracts or pure compounds

451

were accompanied by lower leukocyte infiltration and reduced atherosclerotic lesions in the

452

aorta of these mice. Furthermore, using a microarray-based holistic approach that allows the

A similar action of anthocyanin metabolites has also been shown in few studies

10,71,72,77–

Bharat et al. recently reported the reduced expression of genes encoding VCAM-1,

72.

Similarly, VCAM1 and IL6 downregulation were observed in TNFα-

Furthermore, different metabolites produced from anthocyanin-rich grape/berry extract by

52,53,57.

10,72,76,78–80.

Similar to in vitro

These changes in the gene expression



Page 20 of 46

453

simultaneous study of the impact on several thousand genes, we have observed the changes in

454

the expression of 1261 genes in the aorta of ApoE−/− mice after 2-week dietary supplementation

455

with anthocyanin-rich bilberry extract

456

implicated in different cellular processes such as inflammation, cell adhesion and

457

transmigration, proposing the multi-targeted mode of action of anthocyanins and their

458

metabolites.

85.

These identified differentially expressed genes are

459

In summation, mechanistic evidence shows that the modulation of expression of different

460

genes implicated in processes such as inflammation, monocyte adhesion and transendothelial

461

migration (Figure 4) seems to be the important mechanism by which anthocyanins and their

462

metabolites mediate the protection against the development of atherosclerosis and associated

463

cardiovascular complications.

464 465

3.4.2. Regulation of cell signaling pathways

466

Expression of genes is regulated at the transcriptional level by transcription factors which

467

activity is controlled by cell signaling pathways that depend on protein phosphorylation. Recent

468

data suggest that the reported nutrigenomic effects of anthocyanins and their metabolites are

469

achieved by their direct binding to the cell signaling proteins, which induce changes in protein

470

function and activation of downstream signaling proteins and transcription factors

471

compounds could affect cell signaling pathways such as the NF-κB and MAPK pathways that

472

play an essential role in the initiation and regulation of inflammatory processes. NF-κB is a

473

transcription factor that is kept in an inactive state in the cytoplasm by binding to the inhibitor

474

of kappa B (IκB). Diverse extracellular stimuli, including oxidized LDL or cytokines such as

475

TNFα, can activate the NF-κB signaling pathway and stimulate the IκB kinase complex that

476

triggers the phosphorylation of IκB, eventually causing its ubiquitination and proteasomal

477

degradation. Activated NF-κB heterodimer then translocates to the nucleus where it stimulates

10.

These


Page 21 of 46


478

the expression of proinflammatory genes (e.g., genes encoding cell adhesion molecules and

479

cytokines), inducing the inflammatory response and promoting the leukocyte adhesion 86. In a

480

few studies, anthocyanin-rich extracts or pure compounds have been shown to suppress

481

oxidized LDL- or cytokine-stimulated NF-κB activation in endothelial cells through the

482

inhibition of IκB phosphorylation and translocation of p65 subunit into the nucleus 76,81,87–89.

483

In our latest work, we reported that physiologically relevant mixtures of anthocyanins and their

484

microbial metabolites reduced the phosphorylation of NF-κB-p65 in TNF-activated

485

endothelial cells

486

encoding inflammatory mediators and adhesion molecules as well as reduced monocyte

487

adhesion and transendothelial migration. Reduced NF-κB activation and downregulation of

488

VCAM1 and ICAM1 expressions were also described in the aorta of ApoE−/− mice and

489

associated with the observed reduction of aortic sinus plaque area following the 20-week

490

dietary supplementation with PCA

491

signaling pathway, a group of serine/threonine protein kinases includes the extracellular signal-

492

regulated kinase (ERK), p38 and c-Jun N-terminal kinase (JNK) that play an important role in

493

many cellular processes including inflammation, proliferation and apoptosis

494

studies, the anthocyanin-mediated decrease in the expression of proinflammatory genes in

495

activated endothelial cells has been associated with their ability to reduce JNK, p38 or ERK1/2

496

phosphorylation

497

and MAPK signaling pathways might present the key targets of the action of anthocyanins and

498

their metabolites, mediating their protective effects against CVD.

10.

These effects were accompanied by changes in the expression of genes

10,87,90.

52.

Anthocyanins have been shown to affect MAPK

86.

In a few

The above findings suggest that the inflammation-associated NF-κB

499

Several studies have reported that anthocyanins can also act on nuclear factor erythroid 2-

500

related factor 2 (Nrf2) pathways. Nrf2 is a transcription factor that regulates the activity of

501

many genes involved in cellular protection against impaired redox balance. It is bound to the

502

cytoskeletal-associated protein Kelch-like ECH-associated protein-1, which represses its 21 ACS Paragon Plus Environment


Page 22 of 46

503

transcriptional activity. Upon stimulation, Nrf2 dissociates from its inhibitor and translocates

504

into the nucleus to initiate the expression of antioxidant genes

505

studies have shown that both anthocyanins and their metabolites may increase the nuclear

506

translocation of Nrf2 and upregulate the expression of genes involved in redox signaling (e.g.,

507

genes encoding HO-1, NOQ1 and GLCM), thereby affecting antioxidant defenses in

508

endothelial cells 64,65,68,69. However, it should be noted that despite the in vitro evidence of the

509

action of various polyphenols on Nrf2 signaling, the relevance in humans is yet to be confirmed

510

by mechanistic RCTs

511

might modulate nicotinamide adenine dinucleotide (NAD+), a cofactor shown to act as a key

512

mediator in regulating a broad spectrum of processes including redox balance and vascular

513

repair 93,94. Thus, NAD+ might present the new potential target for future investigation.

92.

91.

Results from few in vitro

Recently, studies on redox signaling suggested that anthocyanins

514 515

3.4.3. Modulation of microRNA (miRNA) expression

516

Modulation of miRNA expression has recently emerged as a possible mechanism by which

517

polyphenols, including anthocyanins, may exert their beneficial health effects. These

518

endogenous, non-coding, single-stranded RNAs of around 22 nucleotides regulate the gene

519

expression on post-transcriptional level depending on the degree of complementarity with their

520

targets. miRNAs that perfectly base-pair with their mRNA targets induce their cleavage, while

521

imperfect binding blocks protein translation. By changing mRNA availability and

522

consequently protein synthesis, miRNAs control both physiological and pathological processes

523

such as the development of CVD 95. Polyphenols have been reported to modulate more than

524

100 miRNAs involved in the regulation of different cellular processes like inflammation and

525

apoptosis

526

expression is still largely unknown. In our in vivo study, we observed that the supplementation

527

of diet with nutritionally relevant doses of anthocyanins or ferulic acid modulated the

95,96.

However, the effect of anthocyanins and their metabolites on miRNA


Page 23 of 46


528

expression of 45 and 28 miRNAs, respectively, in the liver of ApoE−/− mice 96. Recently, we

529

have shown for the first time the capacity of physiologically-relevant concentrations of

530

anthocyanins and their microbial metabolites to significantly modulate miRNA expression in

531

TNF-activated endothelial cells 10. Tested compounds profoundly affected the expression of

532

miRNAs involved in the regulation of endothelial cell permeability and atherosclerosis

533

development. However, more studies with different anthocyanins and their metabolites are

534

necessary to fully establish the modulation of miRNA expression as one of the mechanisms

535

underlying the cardioprotective properties of these compounds.

536 537

3.4.4. Limitations of mechanistic evidence and future directions

538

Due to the unique role of the endothelium as sensor and contributor to disturbed vascular

539

homeostasis and associated diseases, a large body of mechanistic studies have focused their

540

attention to the investigation of the action of anthocyanins and their metabolites on endothelial

541

cell function. These studies propose several molecular mechanisms of action of these

542

compounds that could underlie the reported cardioprotective effects associated with the

543

consumption of anthocyanin-rich sources. Anthocyanins and their metabolites seem to regulate

544

different cellular processes involved in the development of CVD by controlling the activity of

545

cell signaling proteins and transcription factors and modulating the gene and miRNA

546

expressions. The evidence is especially convincing regarding the anti-inflammatory action of

547

anthocyanins and their metabolites, suggesting that the modulation of inflammatory responses

548

and consequent improvements in endothelial function is probably the most important action of

549

these compounds. However, the majority of mechanistic evidence comes from studies that have

550

significant limitations. One of the main constraints is the study design that does not consider

551

the ADME of anthocyanins in the human body. Following the consumption of dietary

552

anthocyanin, these compounds are extensively metabolized in the human body. Anthocyanin 23 ACS Paragon Plus Environment


Page 24 of 46

553

metabolites, especially those derived from microbial metabolism, present the dominant forms

554

in the circulation and might contribute to the beneficial health effects associated with the

555

consumption of anthocyanin-rich sources. However, the large number of the available studies

556

used extracts, aglycones or parent compounds, rather than circulating metabolites, at

557

supraphysiological concentrations (up to 200 µM) and long periods of cell exposure, producing

558

results that lack physiological relevance. Recently, a few studies have tried to overcome this

559

restraint and provide more philological evidence by using circulating microbial metabolites

560

and mixtures of compounds with similar plasma resident time 10,69,72,79,83,84. Still, more reliable

561

in vitro evidence is necessary to support the results reported in vivo. The use of targeted-based

562

approaches, focused on the assessments of a few specific target genes or cell signaling proteins,

563

present another limitation of mechanistic studies. More studies with holistic approaches that

564

allow the simultaneous study of the impact on several thousand targets are needed to create a

565

global picture and adequately address the complexity of the mechanisms underlying the

566

cardioprotective action of anthocyanins and their metabolites. Similarly, the pitfalls of

567

anthocyanin research have been recently discussed in a review that assessed the effect of these

568

compounds on diabetes 97.

569

Another interesting target for exploration of the possible cardioprotective effect of

570

anthocyanins and their metabolites are platelets that also contribute to disturbed vascular

571

homeostasis. Indeed, data from limited in vitro and ex vivo studies suggest that these

572

compounds can modulate platelet function

573

anthocyanins and their microbial metabolites, at physiologically relevant concentrations,

574

reduced platelet activation in response to agonist adenosine diphosphate 100. Additionally, we

575

reported for the first time the capacity of these compounds to reduce platelet aggregation with

576

leukocytes. Increased platelet activation and subsequent aggregation with leukocytes have been

577

shown to promote the leukocyte adhesion and transendothelial migration, further increasing

59,60,98,99.

We have recently shown that several


Page 25 of 46


578

the inflammatory responses and progressing atherosclerosis 37. Still, more studies with different

579

glycosides and metabolites are needed to establish the effects of these compounds on platelet

580

function.

581

In conclusion, despite the growing body of epidemiological clinical and preclinical evidence

582

on the positive role of dietary anthocyanins in preserving cardiovascular health, the underlying

583

mechanisms of their action are still not fully established. More in vitro studies with the

584

physiologically relevant design and integrated, holistic approaches investigating the effect of

585

anthocyanins and their metabolites on the gene, protein and miRNA expressions are needed to

586

completely understand the role of these compounds in CVD prevention. Additionally, special

587

attention should be made in the future to perform well-designed clinical trials to provide solid

588

evidence and ascertain the exact role of anthocyanins in the cardioprotective effect associated

589

with the consumption of anthocyanin-rich foods and decipher underlying molecular

590

mechanisms of action.

591 592

Abbreviations: ADME, absorption, distribution, metabolism and excretion, ApoE−/−,

593

apolipoprotein E knockout; CALD1, caldesmon 1; CALM1, calmodulin 1; CAPN1, calpain 1;

594

CASK, calcium/calmodulin-dependent serine protein kinase; CAV1, caveolin 1; CCL2, C-C

595

Motif Chemokine Ligand 2; CDH5, cadherin 5; CHD, coronary heart disease; CLDN1, claudin

596

1; Cmax, maximal plasma concentration; CRP, C-reactive protein; CVD, cardiovascular

597

diseases; CXCL12, C-X-C motif chemokine ligand 12; CXCL8, C-X-C Motif Chemokine

598

Ligand 8; CYBA, cytochrome B-245 alpha chain; ERK, extracellular signal-regulated kinase;

599

F11R, F11 receptor; F.W., fresh weight; FMD, flow-mediated vasodilatation; GCLC,

600

glutamate-cysteine ligase catalytic subunit, GCLM, glutamate-cysteine ligase regulatory

601

subunit; GIT, gastrointestinal tract; GJA4, gap junction protein alpha 4; GLUT, glucose

602

transporter; HO-1, heme oxygenase-1; ICAM-1, intercellular adhesion molecule 1; IGF1R, 25 ACS Paragon Plus Environment


Page 26 of 46

603

insulin-like growth factor 1 receptor; IKBKB, inhibitor of nuclear factor kappa B kinase

604

subunit beta; IL6, interleukin 6; ITGA5, integrin alpha 5; IκB, inhibitor of kappa B; JAM3,

605

junctional adhesion molecule 3; JNK, c-Jun N-terminal kinase; MCP-1, monocyte

606

chemoattractant protein-1; MI, myocardial infarction; NCF1, neutrophil cytosolic factor 1;

607

NOS2, nitric oxide synthase 2; NOS3, nitric oxide synthase 3; NOX2, NADPH oxidase 2;

608

NOX4, NADPH oxidase 4; NQO1, NADPH quinone oxidoreductase 1; Nrf2, nuclear factor

609

erythroid 2-related factor 2; PCA, protocatechuic acid; PWV, pulse wave velocity; RAGE,

610

advanced glycosylation end product-specific receptor; RCTs, randomized controlled trials;

611

RELA, nuclear factor NF-Kappa-B P65 subunit; RHOC, ras homolog family member C; SELE,

612

E-Selectin; SGLT1, sodium-dependent glucose transporter 1; VCAM-1, vascular cell adhesion

613

molecule 1; TLN1, talin 1.

614 615

Conflict of interest

616

The authors declare no competing financial interest.


Page 27 of 46


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5. References

618

(1)

619 620

Biology of Atherosclerosis. Nature 2011, 473 (7347), 317–325. (2)

621 622

Libby, P.; Ridker, P. M.; Hansson, G. K. Progress and Challenges in Translating the

Tang, G. Y.; Meng, X.; Li, Y.; Zhao, C. N.; Liu, Q.; Li, H. Bin. Effects of Vegetables on Cardiovascular Diseases and Related Mechanisms. Nutrients. 2017, p 857.

(3)

Zhao, C.-N.; Meng, X.; Li, Y.; Li, S.; Liu, Q.; Tang, G.-Y.; Li, H.-B.; Zhao, C.-N.;

623

Meng, X.; Li, Y.; et al. Fruits for Prevention and Treatment of Cardiovascular Diseases.

624

Nutrients 2017, 9 (6), 598.

625

(4)

Pérez-Jiménez, J.; Neveu, V.; Vos, F.; Scalbert, A. Systematic Analysis of the Content

626

of 502 Polyphenols in 452 Foods and Beverages: An Application of the Phenol-Explorer

627

Database. J. Agric. Food Chem. 2010, 58 (8), 4959–4969.

628

(5)

Pérez-Jiménez, J.; Fezeu, L.; Touvier, M.; Arnault, N.; Manach, C.; Hercberg, S.; Galan,

629

P.; Scalbert, A. Dietary Intake of 337 Polyphenols in French Adults. Am. J. Clin. Nutr.

630

2011, 93 (6), 1220–1228.

631

(6)

Cassidy, A.; O’Reilly, É. J.; Kay, C.; Sampson, L.; Franz, M.; Forman, J.; Curhan, G.;

632

Rimm, E. B. Habitual Intake of Flavonoid Subclasses and Incident Hypertension in

633

Adults. Am. J. Clin. Nutr. 2011, 93 (2), 338–347.

634

(7)

Krga, I.; Milenkovic, D.; Morand, C.; Monfoulet, L.-E. An Update on the Role of

635

Nutrigenomic Modulations in Mediating the Cardiovascular Protective Effect of Fruit

636

Polyphenols. Food Funct. 2016, 7 (9), 3656–3676.

637

(8)

Tom-Carneiro, J.; Visioli, F. Polyphenol-Based Nutraceuticals for the Prevention and

638

Treatment of Cardiovascular Disease: Review of Human Evidence. Phytomedicine

639

2015, 23 (11), 1145–1174. https://doi.org/10.1016/j.phymed.2015.10.018.

640 641

(9)

Oteiza, P. I.; Erlejman, A. G.; Verstraeten, S. V.; Keen, C. L.; Fraga, C. G. FlavonoidMembrane Interactions: A Protective Role of Flavonoids at the Membrane Surface? 27 ACS Paragon Plus Environment


642 643

Page 28 of 46

Clin. Dev. Immunol. 2005, 12 (1), 19–25. (10)

Krga, I.; Tamaian, R.; Mercier, S.; Boby, C.; Monfoulet, L.-E.; Glibetic, M.; Morand,

644

C.; Milenkovic, D. Anthocyanins and Their Gut Metabolites Attenuate Monocyte

645

Adhesion and Transendothelial Migration through Nutrigenomic Mechanisms

646

Regulating Endothelial Cell Permeability. Free Radic. Biol. Med. 2018, 124, 364–379.

647

(11)

Andersen, Ø. M.; Jordheim, M. Basic Anthocyanin Chemistry and Dietary Sources. In

648

Anthocyanins in Health and Disease; Wallace, T., Giusti, M., Eds.; CRC Press: New

649

York, 2013; pp 13–90.

650

(12)

Fang, J. Bioavailability of Anthocyanins. Drug Metab. Rev. 2014, 46 (4), 508–520.

651

(13)

de Pascual-Teresa, S.; Sanchez-Ballesta, M. T. Anthocyanins: From Plant to Health.

652 653

Phytochem. Rev. 2008, 7 (2), 281–299. (14)

Hager, T. J.; Howard, L. R.; Prior, R. L. Processing and Storage Effects on Monomeric

654

Anthocyanins, Percent Polymeric Color, and Antioxidant Capacity of Processed

655

Blackberry Products. J. Agric. Food Chem. 2008, 56 (3), 689–695.

656

(15)

Howard, L. R.; Brownmiller, C.; Mauromoustakos, A.; Prior, R. L. Improved Stability

657

of Blueberry Juice Anthocyanins by Acidification and Refrigeration. J. Berry Res. 2016,

658

6 (2), 189–201.

659

(16)

Zamora-Ros, R.; Knaze, V.; Luján-Barroso, L.; Slimani, N.; Romieu, I.; Touillaud, M.;

660

Kaaks, R.; Teucher, B.; Mattiello, A.; Grioni, S.; et al. Estimation of the Intake of

661

Anthocyanidins and Their Food Sources in the European Prospective Investigation into

662

Cancer and Nutrition (EPIC) Study. Br. J. Nutr. 2011, 106 (07), 1090–1099.

663

(17)

Sebastian, R. S.; Wilkinson Enns, C.; Goldman, J. D.; Martin, C. L.; Steinfeldt, L. C.;

664

Murayi, T.; Moshfegh, A. J. A New Database Facilitates Characterization of Flavonoid

665

Intake, Sources, and Positive Associations with Diet Quality among US Adults. J. Nutr.

666

2015, 145 (6), 1239–1248. 28 ACS Paragon Plus Environment

Page 29 of 46

667


(18)

668 669

Lila, M. A.; Burton-Freeman, B.; Grace, M.; Kalt, W. Unraveling Anthocyanin Bioavailability for Human Health. Annu. Rev. Food Sci. Technol. 2016, 7 (1), 375–394.

(19)

Czank, C.; Cassidy, A.; Zhang, Q.; Morrison, D. J.; Preston, T.; Kroon, P. a.; Botting,

670

N. P.; Kay, C. D. Human Metabolism and Elimination of the Anthocyanin, Cyanidin-3-

671

Glucoside: A 13C-Tracer Study. Am. J. Clin. Nutr. 2013, 97 (5), 995–1003.

672

(20)

673 674

(1), 3–13. (21)

675 676

Hribar, U.; Ulrih, N. P. The Metabolism of Anthocyanins. Curr. Drug Metab. 2014, 15

Passamonti, S.; Vrhovsek, U.; Mattivi, F. The Interaction of Anthocyanins with Bilitranslocase. Biochem. Biophys. Res. Commun. 2002, 296 (3), 631–636.

(22)

Oliveira, H.; Fernandes, I.; Brás, N. F.; Faria, A.; De Freitas, V.; Calhau, C.; Mateus, N.

677

Experimental and Theoretical Data on the Mechanism by Which Red Wine

678

Anthocyanins Are Transported through a Human MKN-28 Gastric Cell Model. J. Agric.

679

Food Chem. 2015, 63 (35), 7685–7692.

680

(23)

Liu, Z.; Liu, K. The Transporters of Intestinal Tract and Techniques Applied to Evaluate

681

Interactions between Drugs and Transporters. Asian J. Pharm. Sci. 2013, 8 (3), 151–

682

158.

683

(24)

684 685

Faria, A.; Fernandes, I.; Norberto, S.; Mateus, N.; Calhau, C. Interplay between Anthocyanins and Gut Microbiota. J. Agric. Food Chem. 2014, 62 (29), 6898–6902.

(25)

de Ferrars, R.; Czank, C.; Zhang, Q.; Botting, N.; Kroon, P.; Cassidy, A.; Kay, C. The

686

Pharmacokinetics of Anthocyanins and Their Metabolites in Humans. Br. J. Pharmacol.

687

2014, 171 (13), 3268–3282.

688

(26)

Mink, P. J.; Scrafford, C. G.; Barraj, L. M.; Harnack, L.; Hong, C. P.; Nettleton, J. A.;

689

Jacobs, D. R. Flavonoid Intake and Cardiovascular Disease Mortality: A Prospective

690

Study in Postmenopausal Women. Am. J. Clin. Nutr. 2007, 85 (3), 895–909.

691

(27)

McCullough, M. L.; Peterson, J. J.; Patel, R.; Jacques, P. F.; Shah, R.; Dwyer, J. T. 29 ACS Paragon Plus Environment


Page 30 of 46

692

Flavonoid Intake and Cardiovascular Disease Mortality in a Prospective Cohort of US

693

Adults. Am. J. Clin. Nutr. 2012, 95 (2), 454–464.

694

(28)

Cassidy, A.; O’Reilly ÉJ; Liu, L.; Franz, M.; Eliassen, a. H.; Rimm, E. B. High

695

Anthocyanin Intake Is Associated With a Reduced Risk of Myocardial Infarction in

696

Young and Middle-Aged Women. Circulation 2013, 127 (2), 188–196.

697

(29)

Cassidy, A.; Bertoia, M.; Chiuve, S.; Flint, A.; Forman, J.; Rimm, E. B. Habitual Intake

698

of Anthocyanins and Flavanones and Risk of Cardiovascular Disease in Men. Am. J.

699

Clin. Nutr. 2016, 104 (3), 587–594.

700

(30)

Jennings, A.; Welch, A. A.; Fairweather-Tait, S. J.; Kay, C.; Minihane, A. M.;

701

Chowienczyk, P.; Jiang, B.; Cecelja, M.; Spector, T.; Macgregor, A.; et al. Higher

702

Anthocyanin Intake Is Associated with Lower Arterial Stiffness and Central Blood

703

Pressure in Women. Am. J. Clin. Nutr. 2012, 96 (4), 781–788.

704

(31)

Jennings, A.; Welch, A. A.; Spector, T.; Macgregor, A.; Cassidy, A. Intakes of

705

Anthocyanins and Flavones Are Associated with Biomarkers of Insulin Resistance and

706

Inflammation in Women 1,2. J. Nutr. 2014, 144 (16), 202–208.

707

(32)

Chun, O. K.; Chung, S.-J.; Claycombe, K. J.; Song, W. O. Serum C-Reactive Protein

708

Concentrations Are Inversely Associated with Dietary Flavonoid Intake in U.S. Adults.

709

J. Nutr. 2008, 138 (4), 753–760.

710

(33)

Cassidy, A.; Rogers, G.; Peterson, J. J.; Dwyer, J. T.; Lin, H.; Jacques, P. F. Higher

711

Dietary Anthocyanin and Flavonol Intakes Are Associated with Anti-Inflammatory

712

Effects in a Population of US Adults. Am. J. Clin. Nutr. 2015, 102 (1), 172–181.

713

(34)

Luis, Â.; Domingues, F.; Pereira, L. Association between Berries Intake and

714

Cardiovascular Diseases Risk Factors: A Systematic Review with Meta-Analysis and

715

Trial Sequential Analysis of Randomized Controlled Trials. Food Funct. 2018, 9 (2),

716

740–757. 30 ACS Paragon Plus Environment

Page 31 of 46

717


(35)

García-Conesa, M. T.; Chambers, K.; Combet, E.; Pinto, P.; Garcia-Aloy, M.; Andrés-

718

Lacueva, C.; Pascual-Teresa, S. De; Mena, P.; Ristic, A. K.; Hollands, W. J.; et al. Meta-

719

Analysis of the Effects of Foods and Derived Products Containing Ellagitannins and

720

Anthocyanins on Cardiometabolic Biomarkers: Analysis of Factors Influencing

721

Variability of the Individual Responses. Int. J. Mol. Sci. 2018, 19 (3), E694.

722

(36)

Fairlie-Jones, L.; Davison, K.; Fromentin, E.; Hill, A. The Effect of Anthocyanin-Rich

723

Foods or Extracts on Vascular Function in Adults: A Systematic Review and Meta-

724

Analysis of Randomised Controlled Trials. Nutrients 2017, 9 (8), 908.

725

(37)

Konic-Ristic, A.; Kroon, P. A.; Glibetic, M. Modulation of Platelet Function with

726

Dietary Polyphenols. A Promising Strategy in the Prevention of Cardiovascular Disease.

727

Agro FOOD Indusrty Hi Tech 2015, 26 (2), 15–19.

728

(38)

Thompson, K.; Hosking, H.; Pederick, W.; Singh, I.; Santhakumar, A. B. The Effect of

729

Anthocyanin Supplementation in Modulating Platelet Function in Sedentary Population:

730

A Randomised, Double-Blind, Placebo-Controlled, Cross-over Trial. Br. J. Nutr. 2017,

731

118 (5), 368–374.

732

(39)

Thompson, K.; Pederick, W.; Singh, I.; Santhakumar, A. B. Anthocyanin

733

Supplementation in Alleviating Thrombogenesis in Overweight and Obese Population:

734

A Randomized, Double-Blind, Placebo-Controlled Study. J. Funct. Foods 2017, 32,

735

131–138.

736

(40)

Zhang, X.; Zhu, Y.; Song, F.; Yao, Y.; Ya, F.; Li, D.; Ling, W.; Yang, Y. Effects of

737

Purified Anthocyanin Supplementation on Platelet Chemokines in Hypocholesterolemic

738

Individuals: A Randomized Controlled Trial. Nutr. Metab. (Lond). 2016, 13 (1), 86.

739

(41)

Santhakumar, A. B.; Kundur, A. R.; Fanning, K.; Netzel, M.; Stanley, R.; Singh, I.

740

Consumption of Anthocyanin-Rich Queen Garnet Plum Juice Reduces Platelet

741

Activation Related Thrombogenesis in Healthy Volunteers. J. Funct. Foods 2015, 12, 31 ACS Paragon Plus Environment


742 743

Page 32 of 46

11–22. (42)

Hollands, W. J.; Armah, C. N.; Doleman, J. F.; Perez-Moral, N.; Winterbone, M. S.;

744

Kroon, P. A. 4-Week Consumption of Anthocyanin-Rich Blood Orange Juice Does Not

745

Affect LDL-Cholesterol or Other Biomarkers of CVD Risk and Glycaemia Compared

746

with Standard Orange Juice: A Randomised Controlled Trial. Br. J. Nutr. 2018, 119 (4),

747

415–421.

748

(43)

Hassellund, S. S.; Flaa, A.; Sandvik, L.; Kjeldsen, S. E.; Rostrup, M. Effects of

749

Anthocyanins on Blood Pressure and Stress Reactivity: A Double-Blind Randomized

750

Placebo-Controlled Crossover Study. J. Hum. Hypertens. 2012, 26 (6), 396–404.

751

(44)

Curtis, P. J.; Kroon, P. A.; Hollands, W. J.; Walls, R.; Jenkins, G.; Kay, C. D.; Cassidy,

752

A. Cardiovascular Disease Risk Biomarkers and Liver and Kidney Function Are Not

753

Altered in Postmenopausal Women after Ingesting an Elderberry Extract Rich in

754

Anthocyanins for 12 Weeks. J. Nutr. 2009, 139 (12), 2266–2271.

755

(45)

Giordano, L.; Coletta, W.; Tamburrelli, C.; D’Imperio, M.; Crescente, M.; Silvestri, C.;

756

Rapisarda, P.; Reforgiato Recupero, G.; De Curtis, A.; Iacoviello, L.; et al. Four-Week

757

Ingestion of Blood Orange Juice Results in Measurable Anthocyanin Urinary Levels but

758

Does Not Affect Cellular Markers Related to Cardiovascular Risk: A Randomized

759

Cross-over Study in Healthy Volunteers. Eur. J. Nutr. 2012, 51 (5), 541–548.

760

(46)

Milenkovic, D.; Morand, C.; Cassidy, A.; Konic-Ristic, A.; Tomás-Barberán, F.;

761

Ordovas, J. M.; Kroon, P.; De Caterina, R.; Rodriguez-Mateos, A. Interindividual

762

Variability in Biomarkers of Cardiometabolic Health after Consumption of Major Plant-

763

Food Bioactive Compounds and the Determinants Involved. Adv. Nutr. 2017, 8 (4), 558–

764

570.

765 766

(47)

Feliciano, R. P.; Mills, C. E.; Istas, G.; Heiss, C.; Rodriguez-Mateos, A. Absorption, Metabolism and Excretion of Cranberry (Poly)Phenols in Humans: A Dose Response 32 ACS Paragon Plus Environment

Page 33 of 46


767 768

Study and Assessment of Inter-Individual Variability. Nutrients 2017, 9 (3), 268. (48)

Espín, J. C.; González-Sarrías, A.; Tomás-Barberán, F. A. The Gut Microbiota: A Key

769

Factor in the Therapeutic Effects of (Poly)Phenols. Biochem. Pharmacol. 2017, 139,

770

82–93.

771

(49)

Boto-Ordóñez, M.; Urpi-Sarda, M.; Queipo-Ortuño, M. I.; Tulipani, S.; Tinahones, F.

772

J.; Andres-Lacueva, C. High Levels of Bifidobacteria Are Associated with Increased

773

Levels of Anthocyanin Microbial Metabolites: A Randomized Clinical Trial. Food

774

Funct. 2014, 5 (8), 1932–1938.

775

(50)

Moreno-Indias, I.; Sánchez-Alcoholado, L.; Pérez-Martínez, P.; Andrés-Lacueva, C.;

776

Cardona, F.; Tinahones, F.; Queipo-Ortuño, M. I. Red Wine Polyphenols Modulate

777

Fecal Microbiota and Reduce Markers of the Metabolic Syndrome in Obese Patients.

778

Food Funct. 2016, 7 (4), 1775–1787.

779

(51)

Mauray, A.; Milenkovic, D.; Besson, C.; Caccia, N.; Morand, C.; Michel, F.; Mazur, A.;

780

Scalbert, A.; Felgines, C. Atheroprotective Effects of Bilberry Extracts in Apo E-

781

Deficient Mice. J. Agric. Food Chem. 2009, 57 (23), 11106–11111.

782

(52)

Wang, D.; Wei, X.; Yan, X.; Jin, T.; Ling, W. Protocatechuic Acid, a Metabolite of

783

Anthocyanins, Inhibits Monocyte Adhesion and Reduces Atherosclerosis in

784

Apolipoprotein E-Deficient Mice. J. Agric. Food Chem. 2010, 58 (24), 12722–12728.

785

(53)

Wu, X.; Kang, J.; Xie, C.; Burris, R.; Ferguson, M. E.; Badger, T. M.; Nagarajan, S.

786

Dietary Blueberries Attenuate Atherosclerosis in Apolipoprotein E-Deficient Mice by

787

Upregulating Antioxidant Enzyme Expression. J. Nutr. 2010, 140 (9), 1628–1632.

788

(54)

Suh, J.-H.; Romain, C.; González-Barrio, R.; Cristol, J.-P.; Teissèdre, P.-L.; Crozier, A.;

789

Rouanet, J.-M. Raspberry Juice Consumption, Oxidative Stress and Reduction of

790

Atherosclerosis Risk Factors in Hypercholesterolemic Golden Syrian Hamsters. Food

791

Funct. 2011, 2 (7), 400–405. 33 ACS Paragon Plus Environment


792

(55)

Page 34 of 46

Bhaswant, M.; Shafie, S. R.; Mathai, M. L.; Mouatt, P.; Brown, L. Anthocyanins in

793

Chokeberry and Purple Maize Attenuate Diet-Induced Metabolic Syndrome in Rats.

794

Nutrition 2017, 41, 24–31.

795

(56)

Rodriguez-Mateos, A.; Ishisaka, A.; Mawatari, K.; Vidal-Diez, A.; Spencer, J. P. E.;

796

Terao, J. Blueberry Intervention Improves Vascular Reactivity and Lowers Blood

797

Pressure in High-Fat-, High-Cholesterol-Fed Rats. Br. J. Nutr. 2013, 109, 1746–1754.

798

(57)

Joo, H. K.; Choi, S.; Lee, Y. R.; Lee, E. O.; Park, M. S.; Park, K. B.; Kim, C. S.; Lim,

799

Y. P.; Park, J. T.; Jeon, B. H. Anthocyanin-Rich Extract from Red Chinese Cabbage

800

Alleviates Vascular Inflammation in Endothelial Cells and Apo E−/−mice. Int. J. Mol.

801

Sci. 2018, 19 (3), E816.

802

(58)

Jiang, Y.; Dai, M.; Nie, W. J.; Yang, X. R.; Zeng, X. C. Effects of the Ethanol Extract

803

of Black Mulberry (Morus Nigra L.) Fruit on Experimental Atherosclerosis in Rats. J.

804

Ethnopharmacol. 2017, 200, 228–235.

805

(59)

Yang, Y.; Shi, Z.; Reheman, A.; Jin, J. W.; Li, C.; Wang, Y.; Andrews, M. C.; Chen, P.;

806

Zhu, G.; Ling, W.; et al. Plant Food Delphinidin-3-Glucoside Significantly Inhibits

807

Platelet Activation and Thrombosis: Novel Protective Roles against Cardiovascular

808

Diseases. PLoS One 2012, 7 (5), e37323.

809

(60)

Yao, Y.; Chen, Y.; Adili, R.; McKeown, T.; Chen, P.; Zhu, G.; Li, D.; Ling, W.; Ni, H.;

810

Yang, Y. Plant-Based Food Cyanidin-3-Glucoside Modulates Human Platelet

811

Glycoprotein VI Signaling and Inhibits Platelet Activation and Thrombus Formation. J.

812

Nutr. 2017, 147 (10), 1917–1925.

813

(61)

Ahmet, I.; Spangler, E.; Shukitt-Hale, B.; Joseph, J. A.; Ingram, D. K.; Talan, M.

814

Survival and Cardioprotective Benefits of Long-Term Blueberry Enriched Diet in

815

Dilated Cardiomyopathy Following Myocardial Infarction in Rats. PLoS One 2009, 4

816

(11), e7975. 34 ACS Paragon Plus Environment

Page 35 of 46

817


(62)

818 819

Hadi, H. A. R.; Carr, C. S.; Al Suwaidi, J. Endothelial Dysfunction: Cardiovascular Risk Factors, Therapy, and Outcome. Vasc. Health Risk Manag. 2005, 1 (3), 183–198.

(63)

Tsuda, T.; Shiga, K.; Ohshima, K.; Kawakishi, S.; Osawa, T. Inhibition of Lipid

820

Peroxidation and the Active Oxygen Radical Scavenging Effect of Anthocyanin

821

Pigments Isolated from Phaseolus Vulgaris L. Biochem. Pharmacol. 1996, 52 (7), 1033–

822

1039.

823

(64)

Cimino, F.; Speciale, A.; Anwar, S.; Canali, R.; Ricciardi, E.; Virgili, F.; Trombetta, D.;

824

Saija, A. Anthocyanins Protect Human Endothelial Cells from Mild Hyperoxia Damage

825

through Modulation of Nrf2 Pathway. Genes Nutr. 2013, 8 (4), 391–399.

826

(65)

Fratantonio, D.; Speciale, A.; Ferrari, D.; Cristani, M.; Saija, A.; Cimino, F. Palmitate-

827

Induced Endothelial Dysfunction Is Attenuated by Cyanidin-3-O-Glucoside through

828

Modulation of Nrf2/Bach1 and NF-ΚB Pathways. Toxicol. Lett. 2015, 239 (3), 152–160.

829

(66)

Speciale, A.; Anwar, S.; Canali, R.; Chirafisi, J.; Saija, A.; Virgili, F.; Cimino, F.

830

Cyanidin-3-O-Glucoside Counters the Response to TNF-Alpha of Endothelial Cells by

831

Activating Nrf2 Pathway. Mol. Nutr. Food Res. 2013, 57 (11), 1979–1987.

832

(67)

Sorrenti, V.; Mazza, F.; Campisi, A.; Di Giacomo, C.; Acquaviva, R.; Vanella, L.;

833

Galvano, F. Heme Oxygenase Induction by Cyanidin-3-O-Beta-Glucoside in Cultured

834

Human Endothelial Cells. Mol Nutr Food Res 2007, 51 (5), 580–586.

835

(68)

Ma, Z.-C.; Hong, Q.; Wang, Y.-G.; Tan, H.-L.; Xiao, C.-R.; Liang, Q.-D.; Zhang, B.-

836

L.; Gao, Y. Ferulic Acid Protects Human Umbilical Vein Endothelial Cells from

837

Radiation Induced Oxidative Stress by Phosphatidylinositol 3-Kinase and Extracellular

838

Signal-Regulated Kinase Pathways. Biol. Pharm. Bull. 2010, 33 (1), 29–34.

839

(69)

Tang, J. S.; Vissers, M. C. M.; Anderson, R. F.; Sreebhavan, S.; Bozonet, S. M.;

840

Scheepens, A.; Melton, L. D. Bioavailable Blueberry-Derived Phenolic Acids at

841

Physiological Concentrations Enhance Nrf2-Regulated Antioxidant Responses in 35 ACS Paragon Plus Environment


842 843

Page 36 of 46

Human Vascular Endothelial Cells. Mol. Nutr. Food Res. 2018, 62 (5), 1–34. (70)

Chen, C. Y.; Yi, L.; Jin, X.; Zhang, T.; Fu, Y. J.; Zhu, J. D.; Mi, M. T.; Zhang, Q. Y.;

844

Ling, W. H.; Yu, B. Inhibitory Effect of Delphinidin on Monocyte-Endothelial Cell

845

Adhesion Induced by Oxidized Low-Density Lipoprotein via ROS/P38MAPK/NF-ΚB

846

Pathway. Cell Biochem. Biophys. 2011, 61 (2), 337–348.

847

(71)

Toma, L.; Sanda, G. M.; Niculescu, L. S.; Deleanu, M.; Stancu, C. S.; Sima, A. V.

848

Caffeic Acid Attenuates the Inflammatory Stress Induced by Glycated LDL in Human

849

Endothelial Cells by Mechanisms Involving Inhibition of AGE-Receptor, Oxidative,

850

and Endoplasmic Reticulum Stress. BioFactors 2017, 43 (5), 685–697.

851

(72)

Bharat, D.; Cavalcanti, R. R. M.; Petersen, C.; Begaye, N.; Cutler, B. R.; Costa, M. M.

852

A.; Ramos, R. K. L. G.; Ferreira, M. R.; Li, Y.; Bharath, L. P.; et al. Blueberry

853

Metabolites Attenuate Lipotoxicity-Induced Endothelial Dysfunction. Mol. Nutr. Food

854

Res. 2018, 62 (2).

855

(73)

Petersen, C.; Bharat, D.; Cutler, B. R.; Gholami, S.; Denetso, C.; Mueller, J. E.; Cho, J.

856

M.; Kim, J. S.; Symons, J. D.; Anandh Babu, P. V. Circulating Metabolites of

857

Strawberry Mediate Reductions in Vascular Inflammation and Endothelial Dysfunction

858

in Db/Db Mice. Int. J. Cardiol. 2018, 263, 111–117.

859

(74)

Lazzè, M. C.; Pizzala, R.; Perucca, P.; Cazzalini, O.; Savio, M.; Forti, L.; Vannini, V.;

860

Bianchi, L. Anthocyanidins Decrease Endothelin-1 Production and Increase Endothelial

861

Nitric Oxide Synthase in Human Endothelial Cells. Mol. Nutr. Food Res. 2006, 50 (1),

862

44–51.

863

(75)

Paixão, J.; Dinis, T. C. P.; Almeida, L. M. Malvidin-3-Glucoside Protects Endothelial

864

Cells up-Regulating Endothelial NO Synthase and Inhibiting Peroxynitrite-Induced NF-

865

KB Activation. Chem. Biol. Interact. 2012, 199 (3), 192–200.

866

(76)

Ferrari, D.; Cimino, F.; Fratantonio, D.; Molonia, M. S.; Bashllari, R.; Busà, R.; Saija, 36 ACS Paragon Plus Environment

Page 37 of 46


867

A.; Speciale, A. Cyanidin-3-O-Glucoside Modulates the in Vitro Inflammatory

868

Crosstalk between Intestinal Epithelial and Endothelial Cells. Mediators Inflamm. 2017,

869

2017, 3454023.

870

(77)

Amin, H. P.; Czank, C.; Raheem, S.; Zhang, Q.; Botting, N. P.; Cassidy, A.; Kay, C. D.

871

Anthocyanins and Their Physiologically Relevant Metabolites Alter the Expression of

872

IL-6 and VCAM-1 in CD40L and Oxidized LDL Challenged Vascular Endothelial

873

Cells. Mol. Nutr. Food Res. 2015, 59 (6), 1095–1106.

874

(78)

Kuntz, S.; Asseburg, H.; Dold, S.; Römpp, A.; Fröhling, B.; Kunz, C.; Rudloff, S.

875

Inhibition of Low-Grade Inflammation by Anthocyanins from Grape Extract in an in

876

Vitro Epithelial-Endothelial Co-Culture Model. Food Funct. 2015, 6 (4), 1136–1149.

877

(79)

Krga, I.; Monfoulet, L. E.; Konic-Ristic, A.; Mercier, S.; Glibetic, M.; Morand, C.;

878

Milenkovic, D. Anthocyanins and Their Gut Metabolites Reduce the Adhesion of

879

Monocyte to TNFα-Activated Endothelial Cells at Physiologically Relevant

880

Concentrations. Arch. Biochem. Biophys. 2016, 599, 51–59.

881

(80)

Chao, P. Y.; Huang, Y. P.; Hsieh, W. Bin. Inhibitive Effect of Purple Sweet Potato Leaf

882

Extract and Its Components on Cell Adhesion and Inflammatory Response in Human

883

Aortic Endothelial Cells. Cell Adhes. Migr. 2013, 7 (2), 237–245.

884

(81)

Speciale, A.; Canali, R.; Chirafisi, J.; Saija, A.; Virgili, F.; Cimino, F. Cyanidin-3-O-

885

Glucoside Protection against TNF-α-Induced Endothelial Dysfunction: Involvement of

886

Nuclear Factor-ΚB Signaling. J. Agric. Food Chem. 2010, 58 (22), 12048–12054.

887

(82)

Ma, Z.-C.; Hong, Q.; Wang, Y.-G.; Tan, H.-L.; Xiao, C.-R.; Liang, Q.-D.; Cai, S.-H.;

888

Gao, Y. Ferulic Acid Attenuates Adhesion Molecule Expression in Gamma-Radiated

889

Human Umbilical Vascular Endothelial Cells. Biol. Pharm. Bull. 2010, 33 (5), 752–758.

890 891

(83)

Warner, E. F.; Smith, M. J.; Zhang, Q.; Raheem, K. S.; O’Hagan, D.; O’Connell, M. A.; Kay, C. D. Signatures of Anthocyanin Metabolites Identified in Humans Inhibit 37 ACS Paragon Plus Environment


Page 38 of 46

892

Biomarkers of Vascular Inflammation in Human Endothelial Cells. Mol. Nutr. Food Res.

893

2017, 1700053.

894

(84)

Kuntz, S.; Kunz, C.; Domann, E.; Würdemann, N.; Unger, F.; Römpp, A.; Rudloff, S.

895

Inhibition of Low-Grade Inflammation by Anthocyanins after Microbial Fermentation

896

in Vitro. Nutrients 2016, 8 (7), 411.

897

(85)

Mauray, A.; Felgines, C.; Morand, C.; Mazur, A.; Scalbert, A.; Milenkovic, D. Bilberry

898

Anthocyanin-Rich Extract Alters Expression of Genes Related to Atherosclerosis

899

Development in Aorta of Apo E-Deficient Mice. Nutr. Metab. Cardiovasc. Dis. 2012,

900

22 (1), 72–80.

901

(86)

902 903

Warboys, C. The Role of Blood Flow in Determining the Sites of Atherosclerotic Plaques. F1000 Med. Rep. 2011, 3 (5), 1–8.

(87)

Yi, L.; Chen, C. Y.; Jin, X.; Zhang, T.; Zhou, Y.; Zhang, Q. Y.; Zhu, J. D.; Mi, M. T.

904

Differential Suppression of Intracellular Reactive Oxygen Species-Mediated Signaling

905

Pathway in Vascular Endothelial Cells by Several Subclasses of Flavonoids. Biochimie

906

2012, 94 (9), 2035–2044.

907

(88)

Xia, M.; Ling, W.; Zhu, H.; Wang, Q.; Ma, J.; Hou, M.; Tang, Z.; Li, L.; Ye, Q.

908

Anthocyanin Prevents CD40-Activated Proinflammatory Signaling in Endothelial Cells

909

by Regulating Cholesterol Distribution. Arterioscler. Thromb. Vasc. Biol. 2007, 27 (3),

910

519–524.

911

(89)

Chao, P. Y.; Lin, K.-H.; Chiu, C.-C.; Yang, Y.-Y.; Huang, M.-Y.; Yang, C.-M.

912

Inhibitive Effects of Mulberry Leaf-Related Extracts on Cell Adhesion and

913

Inflammatory Response in Human Aortic Endothelial Cells. Evidence-based

914

Complement. Altern. Med. 2013, 2013 (Article ID 267217), 14.

915 916

(90)

Xia, M.; Ling, W.; Zhu, H.; Ma, J.; Wang, Q.; Hou, M.; Tang, Z.; Guo, H.; Liu, C.; Ye, Q. Anthocyanin Attenuates CD40-Mediated Endothelial Cell Activation and Apoptosis 38 ACS Paragon Plus Environment

Page 39 of 46


917 918

by Inhibiting CD40-Induced MAPK Activation. Atherosclerosis 2009, 202 (1), 41–47. (91)

Hsieh, C. Y.; Hsiao, H. Y.; Wu, W. Y.; Liu, C. A.; Tsai, Y. C.; Chao, Y. J.; Wang, D.

919

L.; Hsieh, H. J. Regulation of Shear-Induced Nuclear Translocation of the Nrf2

920

Transcription Factor in Endothelial Cells. J. Biomed. Sci. 2009, 16 (1).

921

(92)

922 923

Visioli, F. Xenobiotics and Human Health: A New View of Their Pharma-Nutritional Role. PharmaNutrition 2015, 3 (2), 60–64.

(93)

Wang, X.; Zhang, Z.-F.; Zheng, G.-H.; Wang, A.-M.; Sun, C.-H.; Qin, S.-P.; Zhuang,

924

J.; Lu, J.; Ma, D.-F.; Zheng, Y.-L. The Inhibitory Effects of Purple Sweet Potato Color

925

on Hepatic Inflammation Is Associated with Restoration of NAD+ Levels and

926

Attenuation of NLRP3 Inflammasome Activation in High-Fat-Diet-Treated Mice.

927

Molecules 2017, 22 (8), E1315.

928

(94)

Khoo, H. E.; Azlan, A.; Ismail, A.; Abas, F.; Hamid, M. Inhibition of Oxidative Stress

929

and Lipid Peroxidation by Anthocyanins from Defatted Canarium Odontophyllum

930

Pericarp and Peel Using in Vitro Bioassays. PLoS One 2014, 9 (1), e81447.

931

(95)

932 933

Milenkovic, D.; Jude, B.; Morand, C. MiRNA as Molecular Target of Polyphenols Underlying Their Biological Effects. Free Radic. Biol. Med. 2013, 64, 40–51.

(96)

Milenkovic, D.; Deval, C.; Gouranton, E.; Landrier, J. F.; Scalbert, A.; Morand, C.;

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Mazur, A. Modulation of MiRNA Expression by Dietary Polyphenols in ApoE Deficient

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Mice: A New Mechanism of the Action of Polyphenols. PLoS One 2012, 7 (1), e29837.

936

(97)

Crespo, M. C.; Visioli, F. A Brief Review of Blue- and Bilberries’ Potential to Curb

937

Cardio-Metabolic Perturbations: Focus on Diabetes. Curr. Pharm. Des. 2017, 23 (7),

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983–988.

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(98)

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Rechner, A. R.; Kroner, C. Anthocyanins and Colonic Metabolites of Dietary Polyphenols Inhibit Platelet Function. Thromb. Res. 2005, 116, 327—334.

(99)

Baeza, G.; Bachmair, E.-M.; Wood, S.; Mateos, R.; Bravo, L.; de Roos, B. The Colonic 39 ACS Paragon Plus Environment


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Metabolites Dihydrocaffeic Acid and Dihydroferulic Acid Are More Effective

943

Inhibitors of in Vitro Platelet Activation than Their Phenolic Precursors. Food Funct.

944

2017, 8 (3), 1333–1342.

945

(100) Krga, I.; Vidovic, N.; Milenkovic, D.; Konic-Ristic, A.; Stojanovic, F.; Morand, C.;

946

Glibetic, M. Effects of Anthocyanins and Their Gut Metabolites on Adenosine

947

Diphosphate-Induced Platelet Activation and Their Aggregation with Monocytes and

948

Neutrophils. Arch. Biochem. Biophys. 2018, 645, 34–41.


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949

6. Figure captions

950

Figure 1. Chemical structure of common anthocyanidins. Gly, glycoside.

951 952

Figure 2. Scheme of anthocyanin absorption and metabolism. Acy, anthocyanidins; Gly,

953

glycosides, GLUT-1, glucose transporter 1.

954 955

Figure 3. The graphical representation of the average Cmax values of individual anthocyanins

956

in human plasma reported in bioavailability studies. Data are derived from references presented

957

in Table S1.

958 959

Figure 4. Summary of genes related to endothelial dysfunction which expression has been

960

identified as modulated by anthocyanins and their metabolites in vitro. Red-upregulation,

961

green-downregulation. CALD1, caldesmon 1; CALM1, calmodulin 1; CAPN1, calpain 1;

962

CASK, calcium/calmodulin-dependent serine protein kinase; CAV1, caveolin 1; CCL2, C-C

963

Motif Chemokine Ligand 2; CDH5, cadherin 5; CLDN1, claudin 1; CXCL12, C-X-C motif

964

chemokine ligand 12; CXCL8, C-X-C Motif Chemokine Ligand 8; CYBA, cytochrome B-245

965

alpha chain; F11R, F11 receptor; GJA4, gap junction protein alpha 4; HMOX1, heme

966

oxygenase-1; IGF1R, insulin-like growth factor 1 receptor; IKBKB, inhibitor of nuclear factor

967

kappa B kinase subunit beta; IL6, interleukin 6; ITGA5, integrin alpha 5; JAM3, junctional

968

adhesion molecule 3; NCF1, neutrophil cytosolic factor 1; NOS2, nitric oxide synthase 2;

969

NOS3, nitric oxide synthase 3; NOX2, NADPH oxidase 2; NOX4, NADPH oxidase 4; NQO1,

970

NADPH quinone oxidoreductase 1; Nrf2, nuclear factor erythroid 2-related factor 2; RAGE,

971

advanced glycosylation end product-specific receptor; RELA, nuclear factor NF-Kappa-B P65

972

subunit; RHOC, ras homolog family member C; SELE, E-Selectin; TLN1, talin 1.



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7. Figures

Figure 1


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Figure 2



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Figure 3


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Figure 4



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Table of contents graphics


Anthocyanins: from sources and bioavailability to cardiovascular

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