Aristotelia chilensis: A Possible Nutraceutical or Functional Food

In recent years, both in the market of American nutraceuticals as in Chile, there have been a number of products whose formulation containing Aristotelia chilensis. This fruit and leaves have shown a important range of biological actions, including antioxidants, antiinflammatory and analgesic effects. These effects have led to use these products as complementary therapeutic agents in the treatment of various chronic and degenerative disorders. The fruits of Aristotelia chilensis have a high content of anthocyanins, of indole alkaloids and flavonoids, which makes them an attractive resource to produce antioxidant extracts for use in functional foods or nutraceuticals [1,2].


Introduction
A nutraceutical is a product made from a food, which besides having a nutritional role by providing nutrients and energy, is capable of delivering the body other active ingredients beneficial to health, differing from a functional food in the form of presentation and administration [1].
In recent years, both in the market of American nutraceuticals as in Chile, there have been a number of products whose formulation containing Aristotelia chilensis. This fruit and leaves have shown a important range of biological actions, including antioxidants, antiinflammatory and analgesic effects. These effects have led to use these products as complementary therapeutic agents in the treatment of various chronic and degenerative disorders. The fruits of Aristotelia chilensis have a high content of anthocyanins, of indole alkaloids and flavonoids, which makes them an attractive resource to produce antioxidant extracts for use in functional foods or nutraceuticals [1,2].
The Aristotelia chilensis (Mol.) Stuntz, belongs to the family of Elaeocarpareae, a Chilean native species, popularly known as Maqui.
It is a small tree that does not grow beyond 4-5 meters high, evergreen, with reddish stems, thin, flexible branches. Its bark is smooth and soft. Its leaves are stalked, lanceolate, with serrated edges, measuring 4-9 cm. It has small, whitish flowers arranged in axillary inflorescences. Its fruits are small berries of about 5 mm, bright blue or black color. The Maqui grows wild and is distributed from Limarí in Region IV to Region XI Aysén in Chile. It thrives in moist soil, invading ravines, slopes of hills and forest edges [1][2][3][4].
In Chile, this species is known for the use given to the fruit in the diet. However, little has been used for medicinal purposes [5]. Only native peoples (mainly Mapuches) have used its leaves as a tea for therapeutic purposes in the treatment of diarrhea, acute tonsillitis, throat and mouth ulcers [6].
In recent years there has been an active interest in the berries, which are a group of fruits red, violet or purple, with a high content of polyphenols. Among the highlights are cranberries, myrtle, raspberries, grapes, caulk and also maqui or blackberry [7].

Traditional Medical Purposes
Among the traditional medicinal uses, fruits and juices, as well as Maqui leaves have been used [1,6,8]. Internally, the fruits have been used as a tonic that has antidiarrheal, anti-inflammatory, healing,

Abstract
Aristotelia chilensis (Maqui) is a native berry from Chile that has a wide range of biological actions. It is mainly composed of anthocyanins, indole alkaloids and flavonoids. However, the high concentration of polyphenols has been identified as the main responsible for the anti inflammatory, analgesic and antioxidants effects of their extracts, juices and fruit. The nutraceuticals or functional foods development is an excellent opportunity to make a nutritious and healthy product. This review includes the chemical and nutritional characteristics of Maqui. The pharmacological and biological activities are included in this work. Also shows our need to develop new healthy products digestive, diaphoretic, expectorant, laxative and diuretic properties. Infusions of the leaves can be administered as gargling in inflammation of the oropharyngeal mucosa [8]. The Mapuches used the fruits of maqui externally to treat infected wounds and burn dry leaves as healing [1,6,8].

Nutritional Composition
In 1990 Schmidt-Hebbel et al. published tables of chemical composition of Chilean foods where maqui is included as a native fruit [9]: Schmidt-Hebbel et al. Composición Química de Alimentos Chilenos, 8°Edición. Universidad de Chile. According to Damascos et al. the maqui contains a significant amount of ascorbic acid. Native shrub fruits had higher content of Br, Zn, Co, Cr, Fe, Mo, and Na than those of the exotic naturalized species The fruit nutrient content found in the studied species was similar or higher than other values reported for fruits of temperate and tropical species in the world [10].
anti-inflammatory and mutagenic properties, make them interesting objects of research [2].

Antioxidant activity
Polyphenolic compounds maqui fruit have shown to possess potent antioxidant effects in vitro [19,20]. Maqui juices showed prevent lipid peroxidation induced by copper on human endothelial cells. The mechanism is explained by the effect of polyphenols maqui juice and by their antioxidant effect against oxidative stress [21].
Similarly, combining maqui juice with lemon juice important antioxidant effect against DPPH radicals, superoxide anion and hydroxyl were evidenced. The results suggested that lemon juice enriched with berries could be of potential interest in the design of new drinks with a nutritive related function on health for chronic diseases [22].
The new isotonic blends with lemon and anthocyanins-rich berries showed an attractive colour, especially in maqui samples, which is essential for consumer acceptance. Significantly higher antioxidant and biological effects were determined in the new blends, in comparison with the commercial isotonic beverages [23].
In general, aqueous and methanolic extracts of the fruit of the A. chilensis have shown antioxidant effects on in vitro models and xanthine oxidase inhibiting DPPH, fluctuating in the range of 52.9 and 62.7% respectively [12].
Crude extracts of leaves and fruits also showed antioxidant activity. Their polyphenols inhibit alpha-glucosidase enzyme. This enzyme is essential for the metabolism of carbohydrates and using these crude extracts, can be a useful mechanism in the management of diabetic patients. However, is necessary to make more research for warrants a potential dietary use of this berry in therapies to combat diabetes symptoms [24,25].
The Anthocyanins from A. chilensis improve fasting blood glucose levels and glucose tolerance in obese diabetic mice. Decreased glucose production and enhanced insulin-induced gene expression of glucose-6-phosphatase was observed in liver cells. For other hand, the Delphinidin 3-sambubioside-5-glucoside (D3S5G) was identified as principal anthocyanin of Maqui berry and responsible for these effects on glucose utilization [25,26].
The Aristoteline, aristone, serratoline and hobartinol were isolated from mixture of alkaloids (ALK-MIX). Ursolic acid, friedelin and quercetin 5,3'-dimethyl ether were present in the dichloromethane extract while quercetin 3-O-β-d-glucoside and kaempferol were present in the methanol extract. From aqueous extract (INFU) were isolated protopine, aristoteline and caffeic and ferulic acids. The effects of A. chilensis are here in demonstrated, validating its use in traditional medicine. Protopine was reported for the first time in Elaeocarpaceae for Muñoz in 2011 [12].
Phenolics compounds are well-known antioxidants and their regular ingestion contributes to improve antioxidant status of plasma. Phytochemical findings describe phenolics compounds of leaves in Aristotelia chilensis (Molina) Stuntz, show that their administration in herbal teas may support the prevention of diseases whose etiology lies in the oxidative stress. The extracts obtained with polar solvents showed the stabilizing capacity of free radicals and the antioxidant capacity of plasma which, after taking A. chilensis tea, increased to 30.27% as an average. Herbal tea and extracts from A. chilensis exhibited antioxidant capacity under the described conditions [5]. The methanol extract (MeOH) from mature fruits of Aristotelia chilensis (Mol) Stuntz, showed antioxidant activities and cardioprotective effects on acute ischemia/ reperfusion in rat heart in vivo. On the other hand, the methanolic extract of the fruits was able to prevent theses harmful events in the animal's heart by diminishing lipid oxidation. Antioxidant activities of MeOH were strongly correlated with total polyphenol content. Consistent with this finding, MeOH had the greatest ORAC and FRAP values as percentage of activity. These results show that these fruits could be useful as antioxidant, cardioprotective and nutraceuticals sources (Table 1) [27]. The presence of the compound indole-3-OH showed significant antioxidant effects and may explain the chemopreventive, anti-inflammatory and cardiopreventive activities this fruit [2].

Anti-inflammatory and analgesic activities
Folk medicine attributes to the leaves of this species, analgesic and anti-inflammatory properties to be backed by scientific studies and has also determined that the leaves have antimicrobial and antioxidant activities [3,12,28]. The lack of effective and safe long-term therapies to treat the inflammatory process is an important problem in animal health. Thereby, the discovery of new anti-inflammatory alternatives is imperative. We have determined the anti-inflammatory effects of Aristotelia chilensis fruit concentrate in rats subjected to subplantar acute inflammation induced by carrageenan [29].
A bioguided study was carried out on serial extracts (hexane, dichloromethane, methanol, INFU and crude ALK-MIX). All extracts were evaluated for: a) topical administration against both arachidonic acid and 12-deoxyphorbol-13-decanoate (TPA)-induced inflammation in mice b) per-os administration against inflammation by deltacarrageenan-induced paw oedema in guinea-pigs and c) topical analgesia in tail flick and formalin model and per-os writhing test in mice. Greater anti-inflammatory effects were obtained against TPA with dichloromethane extract and methanol extract (63.9% and 66% respectively). INFU showed the most potent effect (56.2%) against arachidonic acid. Greater effects were obtained in the writhing test hexane and dichloromethane extracts (89.2%) [12].
In the topical analgesia models, all the extracts and ALK-MIX and the methanol extract were the most active (58.2 and 55.2%, respectively). In relation to the tail formalin assay, the methanol extract (74.1%) was the most active [12]. The proposed mechanism of action for analgesic activity of the hexane extracts, is considered which would be modulated by L-arginine serotonergic receptors (5-HT3), adrenergic (a-1), cholinergic-muscarinic (ACh-M) and via/NO (nitric oxide). Instead antinociceptive activity of dichloromethane extract would be modulated by the opioid, serotonergic, adrenergic and nitridergic pathways. In the analgesic effect of methanol extract were involved the opioid systems, adrenergic, muscarinic cholinergic and L-arginine/NO. Alkaloids (Aristone, aristotelone, aristoteline and aristotelinine) demonstrated relax intestinal smooth muscle, explaining its anti-inflammatory and antispasmodic actions [3,28].

Calories
In general these compounds that occur in these Aristotelia species have been considered as the active principles of many anti-inflammatory plants. Thus, many phenolic acids, anthocyanins and flavonoids type have shown inhibitory activities on nitric oxide implicated in physiological and pathological process as chronic inflammation. These finding shows that the anthocyanins, flavonoids and phenolic acids may be responsible of the anti-inflammatory activity of this fruit. Actually, researchers are working in the kinetic of inhibition of these plant extracts and compounds as anti-inflammatory and they are dissecting the sites and mechanism of action as iNOS, Cyclooxygenase (COX), and TNF, among others [14]. Likewise, these extracts were able to reduce the production of nitric oxide (3.7-25.5%) and prostaglandin E2 (9.1-89.1%) and the expression of inducible oxide nitric synthase (9.8-61.8%) and cyclooxygenase-2 (16.6-62.0%) in RAW 264.7 macrophages stimulated by lipopolysaccharide [30].
Anthocyanins have effect during the inflammatory process, since they act by inhibiting the COX. Besides its antioxidant purpose and it inhibits adhesion of leukocytes to endothelial reaction to suppress the expression of molecules VCAM cells. They also inhibit mast cell degranulation and decreased levels of IL-2, IFN-y, and TNF-α [31].
It has been shown that anthocyanins inhibit the secretion of inflammatory cytokines such as IL-8, MCP-1, IL-1B-1 in neutrophil chemotactic cytokine-induced and IL-6 in cell and animal model after an inflammatory stimulus [32,33]. This suggests that it is feasible to use a formulation of dermatological use with A. chilensis to treat inflammatory skin diseases such as rosacea, sunburn and more circumscribed inflammatory lesions [34]. Other studies have suggested that supplementation with anthocyanins in vivo inhibits activation of NF-kB regulates many pro-inflammatory chemokines, cytokines and inflammatory mediators [35].
Anthocyanins exhibit a wide range of antioxidant protection and beneficial therapeutic effects including genomic DNA integrity, cardiovascular protection, neurological protection, anti-inflammatory and anti-cancer and can significantly reduce the production of NO, reducing oxidative stress in the anti-inflammatory process [29].

Others biological activities of general interest
A few years ago, the effects of concentrated of A. chilensis on the expression of COX-2, their signaling pathways and cell viability of colon cancer was assessed. Treatment of Caco-2 cells with A. chilensis for 24 hours reduced the expression of the protein and mRNA of COX-2, and reduced the luciferase activity regulated by NF-ĸB or NFAT.
Treatment of Caco-2 cells for 4 hours with A. chilensis temporarily reduced IĸBα cytoplasmic levels, increased phosphorylation of ERK1/2 and Akt and the expression of c-fos. Also cell viability at concentrations that reduced expression of COX-2 was not affected. These results suggest a potential anticancer and anti-inflammatory effect [36].
On the other hand, the tannins of the fruit of the maqui have astringent properties and anti-diarrheal and the polyphenols of the leaves showed effects anti-hemolytic in vitro [24].
An experimental study evaluated the neuroprotective potential of an extract made from maqui polyphenols in Alzheimer's disease model induced by soluble oligomers B-amyloid.
Neurons cultures of rat hippocampus were used and neuroprotection was observed when neurons were incubated with amyloid B (0.5 uM) more maqui extract for 24 hrs. In the mechanism of action was observed a recovery in the frequency transients calcium channel, compared with the control group receiving only B-amyloid (Only B-amiloide=72 ± 3% and B-amiloide+Maqui= 86 ± 2%; n=5). This was positively correlated with observed in spontaneous synaptic activity and the preservation of dendritic branches changes. The antioxidant activity of maqui is primarily responsible for these changes [37]. Alzheimer's disease is characterized by the presence of different types of extracellular and neurotoxic aggregates of B-amyloid. Recently, bioactive compounds extracted from natural sources showing neuroprotective properties have become of interest in brain neuroregeneration. The results suggest that the maqui induces changes in the aggregation kinetics of Aβ producing variations in the nucleation phase. In conclusion, the maqui induces a potent neuroprotection by direct interaction with the B-amyloid aggregates, generating far less toxic species and in this way protecting the neuronal network [37].
The use of methanol extract of the ripe fruit provides cardioprotective activity in models of acute ischemia reperfusion in an in vivo study in rats [27]. Metabolically, has been evaluated in vitro the ability the some extracts of maqui, in reducing adipogenesis and lipid accumulation in adipocytes. In this regard, the phenolic extracts showed inhibit lipid accumulation between 4 to 10.8% in mature adipocytes and between 5.9 and 37.9% in those in differentiation state [1,38]. Concentrated juice of Aristotelia chilensis has a high content of phenolic compounds with antioxidant capacity, which protect both LDL from oxidation and endothelial cells from intracellular oxidative stress, suggesting that Aristotelia chilensis could have anti atherogenic properties [36].
Obesity is characterized by an increase in the infiltration of monocytes into the adipose tissue, causing an inflammatory condition associate with, for example, the development of insulin resistance. Thus, anti-inflammatory based treatments could emerge as a novel and interesting approach. It has been reported that Chilean native fruits maqui (Aristotelia chilensis) and calafate (Berberis microphylla) present high contents of polyphenols, which are known for their antioxidant and anti-inflammatory properties. The extracts of theses fruits present important inhibitory-like features over the inflammatory response of the interaction between adipocytes and macrophages, comprising a potential therapeutic tool against comorbidities associated with obesity development [39].
One study showed the protective effect of topical formulations containing quercetin on damage caused by oxidative stress induced by ultraviolet radiation. Two different formulations were used; a nonionic emulsion with high lipid content and other anionic emulsion with low concentration of lipids. The results showed that the inflammatory damage induced by ultraviolet radiation decreased with both formulations, with better results when using non-ionic formulation rich in lipids [34].

In search of a nutraceutical formulations or a functional food based on A. chilensis
Infusions of its leaves have long been used in the traditional native herbal medicine to treat different ailments. Phytochemical studies of chemical composition of the plant indicate the presence of indolic alkaloids, flavonoids, cianidine glucosides, delfidine, malvidine, petunidine, cumarines and triterpenes. These compounds, particularly the flavonoids, have antioxidant properties. In order to evaluate the mechanisms of its toxicity and their antioxidant properties, the leaves aqueous extracts were induced to interact with human red cells, their isolated unsealed membranes (IUM), and molecular models of the human erythrocyte membrane. Results of the study indicate that aqueous extracts of A. chilensis induced an alteration of human erythrocyte morphology from the normal discoid shape to an echinocytic form, changes that are explained in terms of the extract interaction with the membranes outer phospholipid monolayer .
In recent years, the food industry has shown great interest in developing maqui juice for its extraordinary potential as an antioxidant and functional food. In this sense, the development of a nutraceutical made from A. chilensis seems a logical step to have a product rich in polyphenols, flavonoids and anthocyanins [40].
For the evaluation of products and chemical characterization of the nutrients and healthy compounds, the quality of the product in dry and fresh should consider [41,42].
1) Drying and juice production substantially affected the stability of maqui anthocyanins 2) Stability of cyanidin and delphinidin derivatives differed during processing.
3) Maqui pomace was a valuable source for obtaining polyunsaturated fruit oils. 4) Maqui products exhibited nutritionally relevant mineral and trace element levels.
Increased domestic and international demand of maqui since 2006, increasing from 733 kg to more than 50,000 kg in 2011 (data from Chile Foundation 2012) showing the interest in this fruit [7].
The maqui has been used to make jams, fresh juices with water and sugar or functional foods by providing a number of compounds potentially beneficial for the health and rich in Polyphenolics components. The physical and chemical characteristics of the maqui juice with and without added sugar are showing in the Evidently, the total polyphenols content is not always the same, can be higher or lower depending on the ripeness of the fruit at harvest time, the type of climate, storage conditions and genotype [4,43].
The maqui is characterized as an important source of phenolics compounds [24] and a product with a high content of polyphenols of natural origin, is a healthy product and considered functional [44].
The fruit of the maqui is considered the healthiest berry exotic, because of its particularly high concentration of bioactive polyphenols [27,38] and the use of this fruit rich in polyphenols in preparing of other foods, showed a reduction in the use of artificial additives antioxidants obtaining thus healthier foods [44,45].
The polyphenols, can prevent heart disease, by possessing vasodilatory effects, antibiotics, anti-inflammatory, anti-apoptotic, anti-atherogenic. Also could be prevent colon cancer [36,46], and providing a possible protective effect in preventing diseases of the retina [47]. Regardless of the course to follow; develop functional foods or nutraceuticals, the Maqui is a super food capable of providing innovative alternatives in food and natural product for health.  Table 2: Physical-chemical analysis obtained maqui juice with and without sugar (40).

Parameter
Juice with sugar Juice without sugar