2015
NPC Natural Product Communications
Activities of Tannins – From In Vitro Studies to Clinical Trials
Elwira Sieniawska
Department of Pharmacognosy with Medicinal Plant Unit, Medical University of Lublin, Lublin 20-093, Poland
elwira.sieniawska@gmail.com
Received: July 20th, 2015; Accepted: August 30th, 2015
Tannins are considered as valuable plant secondary metabolites providing many benefits for human health. In this review information was gathered about
bioactivity in vitro and in vivo, as well as about conducted clinical trials. The literature research was based on ScienceDirect, Scopus, and Cochrane databases
and presents a wide range of tested activities of tannins. The described clinical trials verify laboratory tests and show the effective health benefits taken from
supplementation with tannins.
Keywords: Tannins, Bioactivity in vitro, Bioactivity in vivo, Clinical trials,
Introduction
Tannins are polyphenolic secondary metabolites of higher plants
and are both galloyl esters and their derivatives, in which galloyl
moieties or their derivatives are attached to a variety of polyol-,
catechin- and triterpenoid cores, or they are oligomeric and
polymeric proanthocyanidins that can possess different interflavanyl
coupling and substitution patterns [1]. The classical concept for
tannins classification is based on their resistance to hydrolysis
caused by hot water or tannases. Hydrolysable tannins encompass
polyesters of gallic and hexahydroxydiphenic acid (gallotannins and
ellagitannins, respectively), whereas condensed tannins include
oligomers and polymers composed of flavan-3-ol nuclei
(proanthocyanidins) [2]. In the new approach tannins are divided
according to their structural characteristics into four major groups:
gallotannins, ellagitannins, complex tannins and condensed tannins
[1]. Another important classification of tannins is related to stability
of the chemical structure. Tannins and polyphenols related to
tannins can be categorized as polyphenols of constant chemical
structure (Type A) and polyphenols of variable composition (Type
B). Constancy in the chemical structure and composition is essential
for characterizing the biological and pharmacological properties of
any target compound [3].
Bioactivity of tannins
There are a lot of epidemiological data which suggest that tannins
are useful in external treatment of skin inflammation and injuries
and that intake of tannins may prevent the onset of chronic diseases
[4]. Biological effects of tannins have been extensively studied
using various in vitro or animal models, however, clinical data on
humans is still limited to several plant extracts. Tannins may exert
their biological effects in two different ways: as an unabsorbable,
complex structure with binding properties which may produce local
effects in the gastrointestinal tract (antioxidant, radical scavenging,
antimicrobial, antiviral, antimutagenic, and antinutrient), or as
absorbable tannins (low-molecular-weight) and absorbable
metabolites from colonic fermentation of tannins that may produce
systemic effects in various organs [4].
Bioactivity in vitro
Antioxidant and radical scavenging properties: Tannins are known
to inhibit lipid peroxidation in vitro and have the ability to scavenge
free radicals important in cellular prooxidant states. Most of the
activities of tannins, including their free radical-scavenging
Vol. 10
No. 11
1877 - 1884
capacity, largely depend on their structure and degree of
polymerization [5,6]. Lipid peroxidation in rat liver mitochondria
stimulated by adenine 5'-diphosphate and ascorbic acid was most
potently inhibited by the ellagitannin pedunculagin (Figure 1), a
molecule possessing two hexahydroxydiphenyl groups. Complex
tannins, such as epigallocatechingallate (EGCG) (Figure 2), also
displayed potent activity. Lipid peroxidation caused by the xanthin–
xanthine oxidase system in the cell membranes of mouse eye lens,
was strongly inhibited by several tannins among which were the
hydrolysable tannins pentagalloylglucose (Figure 3) and geraniin
(Figure 1) [5]. Highly condensed kaki-tannin significantly inhibited
the auto-peroxidation of lipid in the liver homogenates from healthy
mice and in the induced liver homogenate lipid oxidation model.
What is more, it was estimated that kaki-tannin was 100-times more
active as an inhibitor of autooxidation or induced oxidation than
grape seed proanthocyanidins [7]. However, procyanidins B1 and
B3 (Figure 4) exhibited stronger antioxidant activity than ascorbic
acid and α-tocopherol for linoleic acid in aqueous systems [8]. An
increase in the antiradical power is observed with the degree of
polymerization of proanthocyanidins up to seven [16]. Potent in
vitro scavenging effects were exhibited also in the 2,2-diphenyl-1-
picrylhydrazyl (DPPH) test by the hydrolysable tannins:
pentagalloyl-glucose (Figure 3), tellimagrandins I and II,
pedunculagin, isoterchebin, mallotusinic acid, geraniin and
chebulinic acid (Figure 1), whereas the effects of low molecular
weight polyphenols, except for EGCG and epicatechingallate
(ECG), were low [5].
Antimicrobial activity: Tannins seem to affect bacterial growth in
several ways, such as inhibition of extracellular microbial enzymes,
deprivation of the substrates required for microbial growth or direct
action on microbial metabolism through inhibition of oxidative
phosphorylation [9]. Complexation of metal ions in the bacterial
growth environment by tannins could also be a mechanism of their
antimicrobial action [3].
Antibacterial activity of tannins has been described for several
human gastrointestinal pathogens. Ellagitannins proved to be strong
inhibitors of Staphylococcus bacteria, Candida albicans and
Campylobacter jejuni [4]. Tea catechins have been tested for
antibacterial activity against Helicobacter pylori. Among several
compounds: (+)catechin (C), (−)epicatechin (EC), (−)epigallo-
catechin (EGC) (Figure 2), ECG and EGCG; only EGCG and ECG
1878 Natural Product Communications Vol. 10 (11) 2015
Figure 1: Gallotannins.
Figure 2: Elagitannins.
exhibited antibacterial activity against H. pylori at the 100 µg/mL
concentration level [10]. However, in another study catechin has
been shown to display a dose and time dependent growth inhibitory
effect (as measured either in liquid culture medium or in semisolid
agar plates) for H. pylori and Escherichia coli [11,12]. Funatogawa
and coworkers, who studied a wide range of tannins against H.
pylori, described that hydrolysable tannins and acid-treated
hydrolysable tannins demonstrated promising activity. Hydrolysable
tannin monomers revealed stronger activity than oligomers and
catechins. Procyanidins demonstrated minimal antibacterial activity
against H. pylori, with a few exceptions. Significant activity of
hydrolysable tannins may be attributed to their higher water-
solubility. What is more, acid-treatment hardly affected the
antibacterial activity of hydrolysable tannins in vitro, suggesting
that they can work in the acidic gastric environment. Hydrolysable
tannins did not affect the viability of gastric epithelial cells and may
have a narrow antibacterial spectrum since they demonstrated no
antibacterial activity against E. coli, which is a normal inhabitant of
the human intestinal tract [13]. Gallotannins were described as
active inhibitors of water-insoluble glucan synthesis leading to
bactericidal effects on Streptococcus mutans, S. salivarius, and
Actinomyces viscosus [14,15].
Sieniawska
Figure 3: Monomeric units and complex tannins.
Figure 4: Condensed tannins.
Flavan-3-ol monomers such as (+)catechin and EC may be capable
of influencing the large intestinal bacterial population, even in the
presence of other nutrients, such as carbohydrates and proteins. It
was found that (+)catechin has regulatory properties because it
significantly inhibited growth of Clostridium histolyticum and
enhanced growth of E. coli and members of the Clostridium
coccoides – Eubacterium rectale group. The growth of
Bifidobacterium and Lactobacillus spp. remained relatively
unaffected [16, 17].
The synergistic effects of ellagitannins and antibiotics against
antibiotic-resistant bacteria are one of the most noticeable
antimicrobial activities of tannins. Several dimeric tannins showed
anti-bacterial activity against methicillin-resistant Staphylococcus
aureus (MRSA) and lowered the resistance to oxacillin of this
microorganism. The complex tannin theasinensin A (Figure 2)
caused a noticeable reduction in MIC values of oxacillin for the
MRSA strains. Oxacillin in the presence of theasinensin A
completely inhibited cell growth over 10 hours. Similar reduction of
resistance to oxacillin was observed with the tea polyphenol
(-)-epicatechingallate, procyanidins B3 and B4 and for the
hydrolysable tannins tellimagrandin I, rugosin B and corilagin
(Figure 1). Theasinensin A was also effective in reducing the MIC
values of the other β-lactams, penicillin G and ampicillin, as well as
an aminoglycoside antibiotic, streptomycin. Two procyanidins, B3
and B4 (Figure 4), lowered the MIC of penicillin G, whereas
oenothein B (Figure 1), a macrocyclic ellagitannin dimer
suppressed the antibiotic resistance of MRSA [18, 19].
Interesting antimicrobial activity known for cranberry juice has
been associated with inhibition of E. coli adherence to human
uroepithelium [20]. This activity is attributed to cranberry
Bioactivities of Tannins
proanthocyanidins with unique molecular structure characterized by
a repetition of a catechin unit with one or more A-type linkages. A
bioactive proanthocyanidin metabolite is present in urine, or
properties of the urine are altered by the proanthocyanidins in such
a way that adhesion is inhibited [21].
Antiviral activity: Tannins have the ability to interact with multiple
viral targets. It has been demonstrated by radiolabelled virus
particles of Herpes simplex that the antiviral effects of hydrolysable
and galloylated condensed tannins were due to inhibition of virus
adsorption [22]. The antiherpetic activities of hydrolysable tannins
were dependent on the number of either galloyl or
hexahydroxydiphenoyl groups, while those of condensed tannins
increased with the degree of condensation [23]. Potent anti-human
immune-deficiency virus (HIV) activities were found for the
dimeric ellagitannins oenothein B, coriariin A, and agrimoniin
(Figure 1). Anti-HIV activity was at least partly mediated by
adsorption inhibition [18]. Other authors have suggested that the
inhibition in adsorption may be derived from its binding to
components of the viral envelope (in influenza A virus,
parainfluenza virus, herpes virus type 1 and 2 and hepatitis A),
resulting in inhibition of viral attachment and penetration of the
plasma membrane [24].
The ellagitannin casuarinin (Figure 1) was also found to possess
anti-herpes virus activity in inhibiting viral attachment to cells and
viral penetration, and also disturbing the late event(s) of infection.
[25]. Other hydrolysable tannins, chebulagic acid and punicalagin
were effective in abrogating infection by human cytomegalovirus
(HCMV), hepatitis C virus (HCV), dengue virus (DENV), measles
virus (MV), and respiratory syncytial virus (RSV), at micromolar
concentrations and in dose-dependent manners without significant
cytotoxicity. Moreover, these natural compounds inhibited viral
attachment, penetration, and spread to different degrees for each
virus. Specifically, the tannins blocked all these steps of infection
for HCMV, HCV, and MV [26]. Hydrolysable tannins possess
remarkable HCV protease inhibitory activities. Casuarinin and
chebulagic acid are non-protein and non-peptidomimetic inhibitors
of HCV protease [27]. It has also been demonstrated that
ellagitannins and several condensed tannins are potent inhibitors of
reverse transcriptase [28].
Cardioprotective activity: Tannins have cardioprotective activity
via stabilization of pericardial tissue [29], inhibition of enzymatic
degradation of elastin [30] and reduction of the calcification of
glutaraldehyde- fixed aortic wall [31]. Hydrolysable tannins, in
particular, have anti-ischemic activity and an endothelium-
dependent vasorelaxant effect through the interplay of different
factors such as cyclooxygenase pathway activation, TNF-alpha
inhibition, endothelial nitric oxide synthase activation, and
scavenging of free radical and reactive oxygen species [32]. It was
shown that extracts and purified tannins of Geum japonicum L.
(hydrolysable tannins) caused NO- and cGMP mediated potent
vasorelaxation in rat aortic rings that had been pre-contracted with
the α1-adrenergic receptor agonist phenylephrine. The phenolic
hydroxyl groups of penta-O-galloyl-β-glucoside are essential for its
vasorelaxant effects, because this structure without the hydroxyl
groups, does not cause vasorelaxation. The G. japonicum extract
markedly reduced arterial blood pressure of hypertensive and
normotensive rats in an NO-dependent manner [33]. Similarly,
tannic acid relaxed precontracted human coronary arteries and rat
aortic rings in an endothelium and NO-dependent manner,
accompanied by an increase in vascular GMP levels [34].
Hydrolysable tannins possess an inhibitory effect on propranolol-
induced negative inotropism, and the relative order of potency of
Natural Product Communications Vol. 10 (11) 2015 1879
the tested hydrolysable tannins (praecoxin, 1-desgalloyl rugosin F)
indicates that the galloyl group in the tannin structure is crucial for
the negative inotropic action [35].
Proanthocyanidins possess long-lasting antihypertensive and
vasorelaxing properties linked to endothelium-related factors in
which nitric oxide is involved [36], and also a significant effect in
the protection of heart against myocardial infarction induced by
isoproterenol [37]. In rabbit pulmonary artery, condensed tannin
prepared from cotton bract endothelium-dependently relaxed the
norepinephrine pre-contracted vessels, presumably via formation of
endothelium-derived relaxing factor [38].
Histamine release inhibition: The inhibitory activity of
hydrolysable tannins on O2-induced histamine release depends on
the number and composition of galloyl groups, HHDP groups and
its analogs, but not simply on the number of phenolic hydroxyl
groups [39]. Geraniin (Figure 1), a typical monomeric hydrolysable
tannin with an HHDP group, a galloyl and a DHHDP group in the
molecule, showed a significant inhibition of K02-induced histamine
release. However, the inhibitory effects of the other monomeric
hydrolysable tannins were less potent. Most of the dimeric
hydrolysable tannins exhibited stronger effects on K02 induced
histamine release. Agrimoniin, euphorbin C and oenothein B
(Figure 1), which have unique structures with several HHDP
groups, the euphorbinoyl group or macroring, respectively, showed
the most potent activity [39].
Cytotoxic activity: Procyanidins have been reported to be more
cytotoxic than monomer flavanols to a variety of human cancer cell
lines. Many studies that have compared the effectiveness of various
monomer flavanols indicate that the presence of a galloyl residue on
the 3 position of the C-ring enhances the cytotoxicity of these
compounds [40]. The potential anticancer properties evaluated for
dimer procyanidins that contain galloyl groups, digalloyl dimer B1
and B2 (Figure 4) esters in a number of human cancer cell lines was
associated with a reduction in cell number and an inhibition of cell
proliferation. Digalloyl dimer procyanidins exerted significantly
higher cytotoxic effects than the structurally related complex
tannins: EC, C, ECG, EGCG, (−)-catechingallate (CG) and
procyanidins B1 and B2 (Figures 2 and 4). These results support the
concept that the incorporation of galloyl groups and the
oligomerization of flavanols enhances the cytotoxic effects of
typical monomer flavanols [40].
In laboratory tests, the catechins present in tea leaves have been
shown to inhibit the growth of cancer cells. Experimental studies
demonstrate that anti-cancer activity is likely to be the result of the
antioxidant activity and the direct binding of green tea polyphenols
to proteins, resulting in the modulation of multiple cellular signaling
pathways [41]. Another way of altering the cancer cells is inhibition
of the release of tumor necrosis factor- α (TNF-α). This activity was
observed for the hydrolysable tannins geraniin and corilagin (Figure
1) in an assay for screening cancer-preventive agents suggesting
that tannins of various structures could prevent cancer [5].
Antitumor activities have been shown in studies using purified
ellagitannins: sanguiin H6 and lambertianin. Moreover, an apple
polyphenols extract rich in proanthocyanidins effectively
suppressed epidermal growth factor receptor phosphorylation,
inhibiting the growth of human colon carcinoma cell line HT29 in
vitro [4].
Antidiabetic activity: Tannins possess antidiabetic potential firstly
due to their ability to lower glucose levels by delaying intestinal
glucose absorption and an insulin-like effect on insulin-sensitive
tissues and secondly by delaying the onset of insulin-dependent
1880 Natural Product Communications Vol. 10 (11) 2015
diabetes mellitus by regulating the antioxidant environment of
pancreatic β-cells [4].The lowering of glucose levels can be
obtained by the inhibition of α-amylase and α-glucosidase activity.
Hydrolysable tannins (HT): vescalagin, acutissimin A/B (Figure 3),
epiacutissimin A/B, grandinin/roburin E, hexagalloyl glucose
(Figure 3) and heptagalloyl glucose have been reported to inhibit α-
amylase in the in vitro human starch digestion model [42]. The type
of enzyme inhibition was mixed non-competitive. Moreover, the
inhibitory activity of HT was significantly enhanced in the absence
of proteins in the food matrix, and was also highly dependent on the
sequence of addition of the enzyme and substrate to HT. Pre-
incubation of α-amylase with HT prior to addition of substrate
resulted in a significantly stronger inhibitory activity [42]. Another
in vitro and in silico investigation of inhibition of human pancreatic
α-amylase showed activity of catechins which followed the order:
(+)-gallocatechin-3-O-gallate > ECG > EGCG > EC > EGC > C.
The galloylated catechins have higher binding affinity with α-
amylase than non- galloylated catechins [43]. It has been suggested
that the interaction between tannins, such as galloylated quinic acid,
and human α-amylase depends on the free hydroxyl groups on the
tannins that are able to participate in hydrogen bonding [44].
Tannins have also been shown to inhibit intestinal α-glucosidase
activity effectively,with Ki values in the same range as that of
synthetic inhibitors (acarbose and voglibose), which are already
being used therapeutically to control non-insulin-dependent diabetes
mellitus [45]. The extent of inhibition of α-glucosidase is related to
the proanthocyanidin content, while α-amylase inhibition is caused
by hydrolysable tannins [46]. An insulin-like effect on insulin-
sensitive tissues has recently been suggested for tannins which act
on cells by modifying or interacting with specific proteins of
important intracellular signaling pathways and hence also affecting
their role in improving hyperglycemia. Grape seed procyanidin
extracts have exhibited insulinomimetic properties in vitro [4,47].
Antiobesity effects: Laboratory studies indicate that the antiobesity
effects of polyphenol-rich diets may be attributed to the ability of
polyphenols to interact, directly or indirectly, with adipose tissues
(preadipocytes, adipose stem cells and immune cells). The
antiobesity potential of green tea catechins, particularly EGCG, has
been shown in cell culture. EGCG (10–100 μM), and EC and EGC
(with lower potencies) induced dose- and time-dependent decreases
in adipocyte viability [48] and cell cycle arrest at the G0/G1 phase
[49]. At lower concentrations (0–10 μM), EGCG induced G2/M
growth arrest in a dose-dependent manner in mature adipocytes
[50]. Concurrently, EGCG (0–400 μM) and, less potently, ECG,
EGC and other catechins, induced apoptosis in murine
preadipocytes [51] and mature adipocytes [52] and increased
caspase-3 activity [51]. Furthermore, EGCG (0.5–10 μM) inhibited
preadipocyte differentiation and, at higher concentrations (50–200
μM), cellular triglyceride (TG) accumulation in adipocytes in a
dose- and time-dependent manner [53]. Also 1,2,3,4,6-penta-O-
galloyl-β-D-glucopyranose (β-PGG) and its natural anomer, α-PGG,
stimulated glucose transport in adipocytes by directly binding to the
insulin receptor. α-PGG did not bind to the insulin-binding site of
the insulin receptor, but to another site located on the α-subunit of
the insulin receptor. The binding of α-PGG to the insulin receptor
displaced insulin from the insulin-binding site [54].
Antiinflammatory and wound healing properties: Ellagitannins
(geraniin, corilagin, furosin) (Figure 1) from aqueous extracts of
Phyllanthus muellerianus have been reported as active compounds
with strong wound-healing properties (stimulating effects against
human keratinocytes and dermal fibroblasts) [55]. Also, for
pentagalloylglucose (main constituent of Paeonia suffruticosa) and
for trigalloyl-glucose and gallic acid from Terminalia chebula
Sieniawska
reports have been published on potential wound healing properties
[56,57]. Strong anti-hyaluronidase and anti-elastase activity was
also described for lythri herba [58] Furthermore, local delivery of
tannic acid prevents collagen matrix degradation by cross-linking
fibrous collagen and inhibiting matrix metalloproteinase activity
[59].
Bioactivity in vivo
In in vivo experiments extracts, juices or fruits with described
beneficial to health activity are mainly used.
Antiulcer activity: Tannins from Mouriri pusa were shown to
exhibit cytoprotective and cicatrizing effects on gastric ulcers in
rats. Reduction in ulcer formationin an acute model and
augmentation of regenerative mucosa were obtained in a 14-day
treatment with 25 mg/kg of the tannin fraction. The tannin fraction
promoted a mechanical barrier that protected the stomach from
ulcer formation and facilitated ulcer healing [60].
Antioxidant activity: Administration of tannin rich cocoa to rats
intoxicated with arsenic significantly protected against liver injury.
The toxic effect of arsenic is mediated through the generation of
reactive oxygen species (ROS) and ROS-driven oxidative stress.
The tannin rich cocoa, administered by oral gavage for 4 weeks in
doses ranging from 30 to 300mg/kg, increased the levels of
superoxide dismutase and glutathione peroxidase activities by
directly scavenging ROS, as well as by inhibiting lipid peroxidation
[61]. Antioxidant effects were also obtained in obese-diabetic rats
during the 4 weeks supplementation with cocoa extracts (600 mg/kg
body weight/day). The decreased plasma oxidative stress biomarker
and increased superoxide dismutase activity were results of the
treatment [62].
Antidiabetic activity: The oral administration of Syzygium cumini
bark extract daily over a period of 45 days (300 mg/kg of body
weight) significantly decreased blood glucose levels and increased
levels of plasma insulin and C-peptide of both normal and diabetic
rats [63]. Kaki-tannin prevented a rise in plasma total cholesterol,
non-HDL cholesterol, triglycerides, and insulin in type 2 diabetic
mice fed a high fat diet. Moreover, intake of kaki-tannin prevented
fatty liver and induced the genes related to cholesterol metabolism
[64].
Antiinflammatory activity: The effect of corilagin (Figure 1) on lung
injury caused by bleomycin exposure was investigated in an animal
model of pulmonary fibrosis. Corilagin abrogated bleomycin-
induced lung fibrosis, reduced the number of apoptotic lung cells
and prevented lung epithelial cells from membrane breakdown. The
release of pro-inflammatory cytokines was reversed significantly by
high-dose corilagin (100 mg/kg i.p) and less dramatically by low
dose (10 mg/kg i.p). This lung-protective effect was attributed to its
anti-oxidative, anti-inflammatory and anti-apoptotic activities, as
well as inhibition of TGF-β1 expression and ECM synthesis [65].
Protecting effects on bone marrow hematopoietic stem cells:
Tannins in Sanguisorba root can effectively protect and treat
cyclophosphamide–induced myelosuppression in mice. The tannin
fraction was administered by oral gavage at a dose of 20 mg/kg for
10 days after ip administration of cyclophosphamide (200 mg/kg).
The mechanism might be related to protecting hematopoietic stem
cells of bone marrow, stimulating hematopoiesis recovery, as well
as preventing the apoptosis of hematopoietic stem cells induced by
cyclophosphamide. The effect may result by promoting cells in the
G2/M phase into the G1 phase, and it could promote the production
of DNA photolyase and repair the damaged DNA after
chemotherapy damage [66].
Bioactivities of Tannins
Inhibition of skin-tumor promotion: The complex tannin,
epigallocatechin gallate (Figure 2) reduced a number of the tumor
bearing mice treated with 7,12-dimethylbenzo[a]anthracene
(DMBA) (a tumor initiator) plus teleocidin (a tumor promoter) to
less than a quarter in the mice treated with combination of tumor
initiator and promoter plus EGCG. The average number of tumors
per mouse in the EGCG-treated mice was also reduced to about 5%
[5]. Inhibition by hydrolysable tannin 1,2,3,4,6-penta-O-galloyl-β-
D-glucose in a two-stage skin carcinogenesis assay using DMBA
plus teleocidin reduced the percentage of tumor- bearing mice to
about 50%, and the average number of tumors per mouse to ca. 33%
[5]. Anti-tumor promoting activity was also described for
tenophyllanin A and alienanin B, a C-glucosidic ellagitannin
monomer and dimer in the two stage mouse skin carcinogenesis
assay with DMBA and 12-O-tetradecanoylphorbol-13-acetate
(TPA). These observations suggest that there may be other tannins
that have anti-tumor promoting activity [5].
Tumor inhibition in the gastrointestinal tract: Inhibition of tumor
promotion in the gastrointestinal tract in a model system of mouse
duodenal carcinogenesis with N-ethyl-N-nitro-N-nitrosoguanidine
by feeding EGCG, resulted in the reduction of the number of tumor-
bearing mice to less than one-third of the control group. The
number of tumors-per-mouse was also reduced to about one-third.
EGCG was incorporated into the cytosol and the nuclei [5]. Apple
proanthocyanidins and red wine proanthocyanidins have also been
demonstrated to antagonize colon carcinoma promotion in vivo. In
F344 rats with azoxymethane-induced colon carcinomas, red wine
polyphenol-treated animals had a consistently lower tumor yield
compared with the control. In animals with drinking water
containing 0.01% apple proanthocyanidinsthe total number of
hyperproliferative crypts and aberrant crypt foci was reduced by
50% showing that apple proanthocyanidins alter intracellular
signaling pathways, polyamine biosynthesis and trigger apoptosis in
tumor cells [4]. A tannin-rich diet may therefore have positive local
health effects in the colon.
Host-mediated antitumor activity of ellagitannin oligomers: The
tumor growth inhibiting effect achieved by administration either
before or after intraperitoneal inoculation of tumor cells was
exhibited by several ellagitannin macrocyclic oligomers, such as
oenotheins A and B, camelliin B and woodfordins C, D, E and F.
The most notable inhibitory effect was found for the macrocyclic
dimers oenothein B and woodfordin C, macrocyclic trimers
oenothein A and woodfordin D, and the macrocyclic tetramer
woodfordin F. The effect is attributable to the immune response of
host animals, as shown by stimulation of interleukin 1 (IL-1)
production from human peripheral macrophages by these
ellagitannin oligomers. The activity of dehydroellagitannins, as
either monomers or dimers, was weak. However acyclic dimers,
such as agrimoniin, coriariin A and C, cornusiin A, hirtellins A and
B, rugosins D and E, isorugosin D and tamarixinin A possessed
some activity. Among the monomers, only tellimagrandin II was
active [5, 18].
Clinical trials
As shown in the previous sections tannins have been widely
investigated in order to find their potential health benefits. Pure
tannins, as well as extracts and beverages, have been subjected to
clinical trials on humans. The influence of tannins on metabolic
syndrome, cardiovascular disease, plasma lipid profiles and
inflammation has been studied.
Green tea: Green tea has long been the beverage and medicine of
choice within Asian countries and has recently sparked the interest
Natural Product Communications Vol. 10 (11) 2015 1881
of Western societies due to its polyphenolic content. The effect of
green tea administration on metabolic syndrome and obesity-related
parameters has been reported in a number of human studies since
1999. Green tea was given to fourteen normal healthy persons
(mean age 30 years) to elucidate its effect on certain constituents of
the metabolic syndrome. Arterial blood flow and cytokine
measurements after taking either a 6 g dose of green tea or a 125-
mg dose of caffeine (amount of caffeine in 6 g of green tea)
revealed that flow-mediated dilation of the brachial artery, a
predictor of cardiovascular disease, was improved with green tea
but not with caffeine alone [67]. One hundred and thirty-two obese
individuals (average BMI 32.2 kg/m2) drank a beverage containing
either 625 mg of green tea catechins or a placebo daily for 3
months. Subjects drinking the green tea extract did not lose more
weight, but they had slightly lower triacylglycerol levels [68]. Sixty
subjects 32 to 73 years of age who had serum hyperglycemia
consumed a tea with 544 mg/day of polyphenols for 2 months in a
crossover study. The individuals who drank the tea showed a
modest but significant lowering of glycated hemoglobin [69].
However, another trial did not show a benefit of green tea on the
metabolic syndrome. Thirty-five individuals (average age 42.5
years) with an average BMI of 36 kg/m2 were provided with green
tea in bags (four cups a day), green tea extract capsules (two a day,
total 800 mg of catechins), or a placebo for 2 months. There were
no differences in parameters of the metabolic syndrome among
groups or in proinflammatory cytokine levels [70]. Some other
studies did not find positive results for obesity-related measures.
For example 12-week green tea supplementation to a low-energy
diet, independent of habitual caffeine intake, had no effect on
measures of body weight or body composition in overweight
women at either 4 weeks or 3 months [71]. Similar non-significant
findings were observed in an EGCG-supplemented group of
overweight/obese postmenopausal women who attended regular
aerobic exercise [72]. Lack of beneficial impact of green tea extract
on obesity was also reported in obese children [73].
Phytopreparations of grape seeds: Several clinical studies
examined the effects of grape seed products on obesity-related
energy expenditure, fat oxidation, plasma lipid profiles, glucose
hemostasis and inflammation.Twenty-seven individuals, 25 to 80
years old, with metabolic syndrome consumed either 150 to 300
mg/day of grape seed extract or a placebo for 1 month. There was a
significant decrease in systolic and diastolic blood pressures in
subjects taking either dose of grape seed extract. The higher was the
baseline of oxidized LDL, the greater the decrease in oxidized LDL
with a 300 mg dose [74]. In another study, forty-four women
(premenopausal, mean age 39.7 years, and postmenopausal, mean
age 58.5 years) consumed 36 g/day of a 182 g lyophilized grape
powder per day (equivalent of 1.5 cups/day of grapes) or a sugar-
sweetened placebo mixed with water for 1 month in a single-
blinded crossover investigation. Pre- and postmenopausal women
who drank a grape beverage showed lower triacylglycerols by 15%
and 6%, respectively. There were no changes in the cytokines TNF-
α, C-reactive protein, or IL-6 [75].
Polyphenol rich cocoa: Several human trials have examined the
effect of polyphenol-rich cocoa on lipid levels. The polyphenols
found in cocoa, augment nitric oxide production [76]. In a recent
study, eight patients with coronary artery disease were given cocoa
drinks containing a 375 mg dose of cocoa polyphenols twice a day
for 1 month. Their brachial arteries had an improved vasodilatory
capacity, and they were able to produce larger numbers of
endothelial progenitor cells. The systolic blood pressure of those
subjects taking the dose of cocoa decreased, which was not seen in
another group that consumed only 9 mg twice a day [77]. Meta-
1882 Natural Product Communications Vol. 10 (11) 2015 Sieniawska

analyses of human trials have concluded that cocoa induces slight ears, high blood pressure, muscle soreness, pain, osteoarthritis,
decrease in total cholesterol and LDL and that coca can lower diabetes, attention deficit-hyperactivity disorder (ADHD), a disease
systolic blood pressure by 4.5 mmHg and diastolic blood pressure of the female reproductive system called endometriosis, menopausal
by 2.5 mmHg [78, 79]. symptoms, painful menstrual periods, erectile dysfunction, and an
eye disease called retinopathy. It is also used for preventing
Cranberry phytopreparations: The meta-analyses of data of all disorders of the heart and blood vessels, including stroke, heart
randomized controlled trials (RCTs) or quasi-RCTs of cranberry disease, and varicose veins. Pycnogenol is used to slow the aging
products for the prevention of urinary tract infections showed that, process, maintain healthy skin, improve athletic endurance, and
compared with placebo, water or no treatment, cranberry products improve male fertility [83].
did not significantly reduce the occurrence of symptomatic urinary
tract infections overall or for any of the subgroups: women with Randomized controlled trials evaluating the effectiveness of
recurrent urinary tract infections; older people; pregnant women; Pycnogenol® in adults or children with any chronic disorder were
children with recurrent urinary tract infections; cancer patients; or included in the meta analysis. Fifteen trials with a total of 791
people with neuropathic bladder or spinal injury. The effectiveness participants have evaluated Pycnogenol® for the treatment of seven
of cranberry was not significantly different to antibiotics for women different chronic disorders. These included asthma (two studies; N
and children. Many studies reported low compliance and high = 86), attention deficit hyperactivity disorder (one study; N = 61),
withdrawal/dropout problems which they attributed to chronic venous insufficiency (two studies; N = 60), diabetes
palatability/acceptability of the products, primarily the cranberry mellitus (four studies; N = 201), erectile dysfunction (one study; N
juice. Most studies of other cranberry products (tablets and = 21), hypertension (two studies; N = 69) and osteoarthritis (three
capsules) did not report how much of the 'active' ingredient the studies; N = 293). Two of the studies were conducted exclusively in
product contained, and, therefore, the products may not have had children; the others involved adults. However, due to small sample
enough potency to be effective [80,81]. size, limited numbers of trials per condition, variation in outcomes
evaluated and outcome measures used, as well as the risk of bias in
Hawthorn extracts: Fourteen clinical trials were used in the meta- the included studies, no definitive conclusions regarding the
analysis of effectiveness of hawthorn extract for treating chronic efficacy or safety of Pycnogenol® are possible [84].
heart failure. In most of the studies, hawthorn was used as an
adjunct to conventional treatment. Ten trials including 855 patients Conclusions: This review shows a wide range of studied activities
with chronic heart failure (New York Heart Association classes I to of tannins and reflects the gap between in vitro and iv vivo testing
III) provided data that were suitable for meta-analysis. For the and clinical trials. This gap is obvious when the physiological
physiological outcome of maximal workload, treatment with processes taking place during tannins ingestion, absorption, and
hawthorn extract was more beneficial than placebo. Exercise metabolism are considered. The chemical properties and molecular
tolerance was significantly increased by hawthorn extract. The mass as well as intestinal bacterial interactions with tannins
pressure-heart rate product, an index of cardiac oxygen influence their bioavailability.However, in vitro and in vivo testing
consumption, also showed a beneficial decrease with hawthorn is a first step to clinical trials. Clinical trials are usually conducted
treatment. Symptoms such as shortness of breath and fatigue using plant extracts rich in tannins or functional food instead of
improved significantly with hawthorn treatment as compared with purified compounds and they show that many laboratory tests often
placebo [82]. do not have confirmation in experiments on humans. The results of
clinical trials strongly depend on the participating subjects, their
Pycnogenol: Pycnogenol is a patented specific blend of health conditions and accompanying diseases. What is more,
procyanidins extracted from the bark of the pine (Pinus pinaster clinical trials investigating the same compounds may produce
Aiton., formerly known as P. maritima Mill.). Pycnogenol is used inconsistent results, but they shed light on the actual usefulness and
for treating circulation problems, allergies, asthma, ringing in the health benefits given by tannins.
References
[1] Khanbabaee K, Van Ree T. (2001) Tannins: Classification and definition. Natural Products Reports, 18, 641–649.
[2] He F, Pan QH, Shi Y, Duan ChQ. (2008) Biosynthesis and genetic regulation of proanthocyanidins in plants. Molecules, 13, 2674-2703.
[3] De Bruyne T, Pieters L, Deelstra H, Vlietinck A. (1999) Condensed vegetable tannins: Biodiversity in structure and biological activities.
Biochemical Systematics & Ecology, 27, 445-459.
[4] Serrano J, Puupponen-Pimi R, Dauer A, Aura AM, Saura-Calixto F. (2009) Tannins: Current knowledge of food sources, intake, bioavailability and
biological effects. Molecular Nutrition & Food Research, 53, 310–329.
[5] Okuda T. (2005) Systematics and health effects of chemically distinct tannins in medicinal plants Phytochemistry, 66, 2012–2031.
[6] Jerez M, Tourio S, Sineiro J, Torres JL, Nuez MJ. (2007) Procyanidins from pine bark: Relationships between structure, composition and
antiradical activity. Food Chemistry, 104, 518–27.
[7] Tian Y, Zou B, Li CM, Yang J, Xu SF, Hagerman AE. (2012) High molecular weight persimmon tannin is a potent antioxidant both ex vivo and in
vivo. Food Research International, 45, 26–30.
[8] Ariga T, Koshiyama I, Fukushima D. (1988) Antioxidative properties of procyanidins B-1 and B-3 from adzuki beans in aqueous system.
Agricultural & Biological Chemistry, 52, 2717–2722.
[9] Scalbert A. (1991) Antimicrobial properties of tannins. Phytochemistry, 30, 3875–3883.
[10] Yilmaz Y. (2006) Novel uses of catechins in foods. Trends in Food Science & Technology, 17, 64–71.
[11] Díaz-Gomez R, Toledo-Araya H, Lopez-Solís R, Obreque-Slier E. (2014) Combined effect of gallic acid and catechin against Escherichia coli.
LWT-Food Science and Technology, 59, 896-900.
[12] Díaz-Gomez R, Lopez-Solís R, Obreque-Slier E, Toledo-Araya H. (2013) Comparative antibacterial effect of gallic acid and catechin against
Helicobacter pylori. LWT-Food Science and Technology, 54, 331-335.
[13] Funatogawa K, Hayashi S, Shimomura H, Yoshida T, Hatano T, Ito H, Hiraii Y. (2004) Antibacterial activity of hydrolysable tannins derived from
medicinal plants against Helicobacter pylori; Microbiology and Immunology, 48, 251–261.
[14] Wu-Yuan CD, Chen CY, Wu RT. (1988) Gallotannins inhibit growth, water insoluble glucan synthesis, and aggregation of mutans streptococci.
Journal of Dental Research, 67, 51–55.
[15] Singh IP, Bharate SB, Bhutani KK. (2005) Anti-HIV natural products. Current Science, 89, 269–290.
Bioactivities of Tannins Natural Product Communications Vol. 10 (11) 2015 1883

[16] Tzonuis X, Vulevic J, Kuhnle GG, George T, Leonczak J, Gibson GR, Kwik-Uribe C, Spencer JP. (2008) Flavanol monomer-induced changes to
the human faecal microflora. British Journal of Nutrition, 99, 782–792.
[17] Cardona F, Andrés-Lacuev C, Tulipani S, Tinahones FJ, Queipo-Ortuño MI. (2013) Benefits of polyphenols on gut microbiota and implications in
human health. Journal of Nutritional Biochemistry, 24, 1415–1422.
[18] Okuda T, Ito H. (2011) Tannins of constant structure in medicinal and food plants - hydrolysable tannins and polyphenols related to tannins.
Molecules, 16, 2191-2217.
[19] Hatano T, Kusuda M, Inada K, Ogawa T, Shiota S, Tsuchiya T, Yoshida T. (2005) Effects of tannins and related polyphenols on methicillin-
resistant Staphylococcus aureus. Phytochemistry, 66, 2047–2055.
[20] Howell AB, Vorsa N, Der Marderosian A, Foo LY. (1998) Inhibition of the adherence of P-fimbriated Escherichia coli to uroepithelial-cell surfaces
by proanthycyanidin extracts from cranberries. New England Journal of Medicine, 339, 1085–1086.
[21] Howell AB, Leahy M, Kurowska E, Guthrie N. (2001) In vivo evidence that cranberry proanthocyanidins inhibit adherence of P-fimbriated E. coli
bacteria to uroepithelial cells. FASEB Journal, 15, A284.
[22] Fukuchi K, Sakagami H, Okuda T, Hatano T, Tanuma S, Kitajima K, Inoue Y, Inoue S, Ichikawa S, Nonoyama M, Konno K. (1989) Inhibition of
herpes simplex virus infection by tannins and related compounds. Antiviral Research, 11, 285–297.
[23] Takechi M, Tanaka Y, Takehara M, Nonaka GI, Nishioka I. (1985) Structure and antiherpetic activity among the tannins. Phytochemistry, 24,
2245–2250.
[24] Ubillas R, Jolad SD, Bruening RC, Kernan MR, King SR, Sesin DF, Barrett M, Stoddart CA, Flaster T, Kuo J, Ayala F, Meza E, Castanel M,
McMekkin D, Rozhon E, Tempesta MS, Barnard D, Huffman J, Smee D, Sidwell R, Soike K, Brazier A, Safrin S, Orlando R, Kenny PT, Berova N,
Nakanishi K. (1994) SP-303 an antiviral oligomeric proantocyanidin from the latex of Croton lechleri (Sangre de Drago). Phytomedicine, 1, 77–
106.
[25] Cheng H, Lin C, Lin T. (2002) Antiherpes simplex virus type 2 activity of casuarinin from the bark of Terminalia arjuna Linn. Antiviral Research,
55, 447–455.
[26] Lin LT, Chen TY, Lin SC, Chun CY, Lin TC, Wang GH, Anderson R, Lin CC, Richardson CD. (2013) Broad-spectrum antiviral activity of
chebulagic acid and punicalagin against viruses that use glycosaminoglycans for entry. BMC Microbiology, 13, 187–202.
[27] Ajala OS, JukovA, MaCM. (2014) Hepatitis C virus inhibitory hydrolysable tannins from the fruits of Terminalia chebula. Fitoterapia, 99, 117–
123.
[28] Kakiuchi N, Kusumoto I, Hattori M, Namba T, Hatano T, Okuda T. (1991) Effect of condensed tannins and related compounds on reverse
transcriptase. Phytotherapy Research, 5, 270–272.
[29] Jastrzebska M, Zalewska-Rejdak J, Wrzalik R, Kocot A, Mroz I, Barwinski B, Turek A, Cwalina B. (2006) Tannic acid-stabilized pericardium
tissue: IR spectroscopy, atomic force microscopy, and dielectric spectroscopy investigations. Journal of Biomedical Materials Research, 78A, 148–
156.
[30] Isenburg JC, Simionescu DT, Vyavahare NR. (2004) Elastin stabilization In cardiovascular implants: improved resistance to enzymatic degradation
by treatment with tannic acid. Biomaterials, 25, 3293–3302.
[31] Isenburg JC, Simionescu DT, Vyavahare NR. (2005) Tannic acid treatment enhances biostability and reduces calcification of glutaraldehyde fixed
aortic wall. Biomaterials, 25, 1237–1245.
[32] Beretta G, Rossoni G, Santagati NA, Facino RM. (2009) Anti-ischemic activity and endothelium-dependent vasorelaxant effect of hydrolysable
tannins from the leaves of Rhus coriaria (Sumac) in isolated rabbit heart and thoracic aorta. Planta Medica, 75, 1482-1488.
[33] Xie YW, Xuc HX, Dongc H, Fiscus RR, But PPH. (2007) Role of nitric oxide in the vasorelaxant and hypotensive effects of extracts and purified
tannins from Geum japonicum. Journal of Ethnopharmacology, 109, 128–133.
[34] Flesch M, Schwarz A, Bohm M. (1998) Effects of red and white wine on endothelium-dependent vasorelaxation of rat aorta and human coronary
arteries. American Journal of Physiology, 275, 1183–1190.
[35] Lee H, Lee JY, Suh MH, Sim SS, Lee M, Kim CJ. (2010) Hydrolysable tannins depress cardiac papillary muscle contraction and propranolol-
induced negative inotropism. Fitoterapia, 81, 820–825.
[36] Magos GA, Mateos JC, Páez E, Fernández G, Lobato C, Márquez C, Enriquez RG. (2008) Hypotensive and vasorelaxant effects of the procyanidin
fraction from Guazuma ulmifolia bark in normotensive and hypertensive rats. Journal of Ethnopharmacology, 117, 58–68.
[37] Karthikeyan K, Bai BR, Devaraj SN. (2007) Cardioprotective effect of grape seed proanthocyanidins on isoproterenol-induced myocardial injury in
rats. International Journal of Cardiology, 115, 326–333.
[38] Russell JA, Rohrbach MS. (1989) Tannin induces endothelium-dependent contraction and relaxation of rabbit pulmonary artery. American Review
of Respiratory Disease, 139, 498–503.
[39] Kanohl R, Hatano T, Ito H, Yoshida T, AkagiM. (2000) Effects of tannins and related polyphenols on superoxide-induced histamine release from
rat peritoneal mast cells. Phytomedicine, 7, 297-302.
[40] Actis-Goretta L, Romanczyk LJ, Rodriguez CA, Kwik-Uribe C, Keen CL. (2008) Cytotoxic effects of digalloyl dimer procyanidins in human
cancer cell lines. Journal of Nutritional Biochemistry, 19, 797–808.
[41] Gadkaria PV, Balaraman M. (2015) Catechins: Sources, extraction and encapsulation.Food and Bioproducts Processing, 93, 122–138.
[42] Tong WY, Wang H, Waisundara VY. (2014) Huang D. Inhibiting enzymatic starch digestion by hydrolysable tannins isolated from Eugenia
jambolana; LWT-Food Science and Technology, 59, 389-395.
[43] Miao M, Jiang H, Jiang B, Li Y, Cui SW, Zhang T. (2014) Structure elucidation of catechins for modulation of starch digestion. LWT-Food Science
and Technology, 57, 188-193.
[44] Kandra L, Gyemant G, Zajacz A, Batta G. (2004) Inhibitory effects of tannin on human salivary α-amylase. Biochemical and Biophysical Research
Communications, 319, 1265–1271.
[45] Matsui T, Ueda T, Oki T, Sugita K, Terahara N, Matsumoto K. (2001) Alpha-glucosidase inhibitory action of natural acylated anthocyanidins.
Survey of natural pigments with potent inhibitory activity. Journal of Agricultural and Food Chemistry, 49, 1948–1951.
[46] McDougall GJ, Shpiro F, Dobson P, Smith P, Blake A, Stewart D. (2005) Different polyphenolic components of soft fruits inhibit α-amylase and
α-glucosidase. Journal of Agricultural and Food Chemistry, 53, 2760–2766.
[47] Pinent M, Blay M, Blade MC, Salvado, MJ, Arola L, Ardevol A. (2004) Grape seed-derived procyanidins have an antihyperglycemic effect in
streptozotocin-induced diabetic rats and insulinomimetic activity in insulin-sensitive cell lines. Endocrinology, 145, 4985-4990.
[48] Moon HS, Chung CS, Lee HG, Kim TG, Choi YJ, Cho CS. (2007) Inhibitory effect of (−)-epigallocatechin-3-gallate on lipid accumulation of 3 T3-
L1 cells. Obesity, 15, 2571–2582.
[49] Hung PF, Wu BT, Chen HC, Chen YH, Chen CL, Wu MH. (2005) Antimitogenic effect of green tea (−)-epigallocatechin gallate on 3 T3-L1
preadipocytes depends on the ERK and Cdk2 pathways. American Journal of Physiology - Cell Physiology, 288, 1094–1108.
[50] Chan CY, Wei L, Castro-Munozledo F, Koo WL. (2011) (−)-Epigallocatechin-3-gallate blocks 3T3-L1 adipose conversion by inhibition of cell
proliferation and suppression of adipose phenotype expression. Life Science, 89, 779–785.
1884 Natural Product Communications Vol. 10 (11) 2015 Sieniawska

[51] Wu BT, Hung PF, Chen HC, Huang RN, Chang HH, Kao YH. (2005) The apoptotic effect of green tea (−)-epigallocatechin gallate on 3 T3-L1
preadipocytes depends on the Cdk2 pathway. Journal of Agricultural and Food Chemistry, 53,5695–5701.
[52] Lin J, Della-Fera MA, Baile CA. (2005) Green tea polyphenol epigallocatechin gallate inhibits adipogenesis and induces apoptosis in 3 T3-L1
adipocytes. Obesity Research, 13, 982–990.
[53] Wang S, Moustaid-Moussa N, Chen L, Mo H, Shastri A, Su R, Bapat P, Kwun I, Shen CL. (2014) Novel insights of dietary polyphenols and
obesity. Journal of Nutritional Biochemistry, 25, 1–18.
[54] Li Y, Kim J, Li J, Liu F, Liu X, Himmeldirk K, Ren Y, Wagner TE, Chen X. (2005) Natural anti-diabetic compound 1,2,3,4,6-penta-O-galloyl-D-
glucopyranose binds to insulin receptor and activates insulin-mediated glucose transport signaling pathway.Biochemical and Biophysical Research
Communications, 336, 430–437.
[55] Agyare C, Lechtenberg M, Deters A, Petereit F, Hensel A. (2011) Ellagitannins from Phyllanthus muellerianus (Kuntze) Exell.:geraniin and furosin
stimulate cellular activity, differentiation and collagen synthesis of human skin keratinocytes and dermal fibroblasts. Phytomedicine, 18, 617–624.
[56] Wang R, Lechtenberg M, Sendker J, Petereit F, Deters A, Hensel A. (2013) Wound healing plants from TCM: in vitro investigations on selected
TCM plants and their influence on human dermal fibroblasts and keratinocytes. Fitoterapia, 84, 308–317.
[57] Singh D, Choi SM, Zo SM, Painuli RM, Kwon SW, Han SS. (2014) Effect of extracts of Terminalia chebula on proliferation of keratinocytes and
fibroblasts cells: an alternative approach for wound healing. Evidenced Based and Complementary and Alternative Medicine, 2014; ID701656.
[58] Piwowarski JP, Kiss AK, Kozłowska-Wojciechowska M. (2011) Anti-hyaluronidase and anti-elastase activity screening of tannin- rich plant
materials used in traditional Polish medicine for external treatment of diseases with inflammatory background. Journal of Ethnopharmacology, 137,
937–941.
[59] Zhang H, Zhu SJ, Wang D, Wei YJ, Hu SS. (2009) Intra myocardial injection of tannic acid attenuates postinfarction remodeling: a novel approach
to stabilize the breaking extracellular matrix. Journal of Thoracic and Cardiovascular Surgery, 137, 216–222.
[60] Vasconcelos PCP, Andreo MA, Vilegas W, Hiruma-Lima CA, Pellizzon CH. (2010) Effect of Mouriri pusa tannins and flavonoids on prevention
and treatment against experimental gastric ulcer. Journal of Ethnopharmacology, 131, 146–153.
[61] Chandranayagam Ch, Veeraraghavan G, Subash A, Vasanthi HR. (2013) Restoration of arsenite induced hepato-toxicity by crude tannin rich
fraction of Theobroma cacao in Sprague Dawley rats. Food Research International, 50, 46–54.
[62] Abbe MJ, Amin I, Chong P, Muhajir H, Hasbullah SK. (2008) Effects of cocoa extract on glucometabolism, oxidative stress, and antioxidant
enzymes in obese-diabetic (ob-db) rats. Journal of Agricultural and Food Chemistry, 56, 7877–7884.
[63] Saravanan G, Leelavinothan P. (2006) Effects of Syzygium cumini bark on blood glucose, plasma insulin and C-peptide in streptozotocin induced
diabetic rats. International Journal of Endocrinology and Metabolism, 4, 96-105.
[64] Matsumoto K, Yokoyama S. (2012) Induction of uncoupling protein-1 and -3 in brown adipose tissue by kaki-tannin in type 2 diabetic NSY/Hos
mice. Food and Chemical Toxicology, 50, 184–190.
[65] Wang Z, Guo QY, Zhang XJ, Li X, Li WT, Ma XT, Ma LJ. (2014) Corilagin attenuates aerosol bleomycin-induced experimental lung injury
International Journal of Molecular Sciences, 15, 9762-9779.
[66] Xiong Z, Yu QN, Zou ZB, He ZH, Zhang SJ, Xu RC, Yang M. (2014) Protective effects of tannins in Sanguisorbae radix on myelosuppression
mice. Chinese Herbal Medicines, 6, 222-227.
[67] Alexopoulos N, Vlachopoulos C, Aznaouridis K, Baou K, Vasiliadou C, Pietri P, Xaplanteris P, Stefanadi E, Stefanadis C. (2008) The acute effect
of green tea consumption on endothelial function in healthy individuals. European Journal of Cardiovascular Prevention & Rehabilitation, 15,
300–305.
[68] Maki KC, Reeves MS, Farmer M, Yasunaga K, Matsuo N, Katsuragi Y, Komikado M, Tokimitsu I, Wilder D, Jones F, Blumberg JB, Cartwright Y.
(2009) Green tea catechin consumption enhances exercise-induced abdominal fat loss in overweight and obese adults. Journal of Nutrition, 139,
264–270.
[69] Fukino Y, Ikeda A, Maruyama K, Aoki N, Okubo T, Iso H. (2008) Randomized controlled trial for an effect of green tea-extract powder
supplementation on glucose abnormalities. European Journal of Clinical Nutrition, 62, 953–960.
[70] Basu A, Du M, Sanchez K, Leyva MJ, Betts NM, Blevins S, Wu M, Aston CE, Lyons TJ. (2011) Green tea minimally affects biomarkers of
inflammation in obese subjects with metabolic syndrome. Nutrition, 27, 206-213.
[71] Diepvens K, Kovacs EM, Nijs IM, Vogels N, Westerterp-Plantenga MS. (2005) Effect of green tea on resting energy expenditure and substrate
oxidation during weight loss in overweight females. British Journal of Nutrition, 94, 1026–1034.
[72] Hill AM, Coates AM, Buckley JD, Ross R, Thielecke F, Howe PR. (2007) Can EGCG reduce abdominal fat in obese subjects? Journal of the
American College of Nutrition, 26, 396–402.
[73] Matsuyama T, Tanaka Y, Kamimaki I, Nagao T, Tokimitsu I. (2008) Catechin safely improved higher levels of fatness, blood pressure, and
cholesterol in children. Obesity, 16, 1338–1348.
[74] Sivaprakasapillai B, Edirisinghe I, Randolph J, Steinberg F, Kappagoda T. (2009) Effect of grape seed extract on blood pressure in subjects with the
metabolic syndrome. Metabolism, 58, 1743–1746.
[75] Zern TL, Wood RJ, Greene C, West KL, Liu Y, Aggarwal D, Shachter NS, Fernandrz ML. (2005) Grape polyphenols exert a cardioprotective effect
in pre- and postmenopausal women by lowering plasma lipids and reducing oxidative stress. Journal of Nutrition, 135, 1911–1917.
[76] Campia U, Panza JA. (2008) Flavanol-rich cocoa a promising new dietary intervention to reduce cardiovascular risk in type 2 diabetes? Journal of
the American College of Cardiology, 51, 2150–2152.
[77] Heiss C, Jahn S, Taylor M, Real WM, Angeli FS, Wong ML, Amabile N, Prasad M, Rassaf T, Ottaviani JI, Mihardja S, Keen CL, Springer ML,
Boyle A., Grossman W, Glantz SA, Schroeter H, Yeghiazarians Y. (2010) Improvement of endothelial function with dietary flavanols in
associated with mobilization of circulating angiogenic cells in patients with coronary artery disease. Journal of the American College of
Cardiology, 56, 218–224.
[78] Ried K, Sullivan TR, Fakler P, Frank OR, Stocks NP. (2012) Effect of cocoa on blood pressure. Cochrane Database Systemic Reviews,
DOI: 10.1002/14651858.CD008893.pub2.
[79] Jia L, Liu X, Bai YY, Li SH, Sun K, He C, Hui R. (2010) Short-term effect of cocoa product consumption on lipid profile: a meta-analysis of
randomized controlled trials. American Journal of Clinical Nutrition, 92, 218–225.
[80] Jepson RG, Williams G, Craig JC. (2012) Cranberries for preventing urinary tract infections Cochrane Database Systemic Reviews,
DOI: 10.1002/14651858.CD001321.pub5.
[81] Barbosa-Cesnik C, Brown MB, Buxton M, Zhang L, DeBusscher J, Foxman B. (2011) Cranberry juice fails to prevent recurrent urinary tract
infection: results from a randomized placebo-controlled trial. Clinical Infection Diseases, 52, 23–30.
[82] Guo R, PittlerMH, Ernst E. (2008) Hawthorn extract for treating chronic heart failure. Cochrane Database Systemic Reviews,
DOI: 10.1002/14651858.CD005312.pub2.
[83] D'Andrea G. (2010) Pycnogenol: A blend of procyanidins with multifaceted therapeutic applications? Fitoterapia, 81, 724–736.
[84] Schoonees A, Visser J, Musekiwa A, Volmink J. (2011) Pycnogenol® (extract of French maritime pine bark) for the treatment of chronic disorders.
Cochrane Database Systemic Reviews,DOI: 10.1002/14651858.CD008294.pub4.