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Blueberries are among the edible fruits that are recognized best for their potential health benefits. The crude extract from Vaccinium corymbosum was assessed in anti-inflammatory and antinociceptive models. The crude hydroalcoholic extract was administered orally at doses of 100, 200 or 300 mg kg (-1) for all the assays. In the carrageenan test, the crude extract reduced rat paw oedema by 9.8, 28.5 and 65.9%, respectively. For the histamine assay, the reductions of oedema were 70.1, 71.7 and 81.9%, respectively. In the myeloperoxidase (MPO) assay, 300 mg kg (-1) crude extract produced a significant inhibition of the MPO activity, at 6 h and 24 h after injection of carrageenan, by 42.8 and 46.2%, respectively. With the granulomatous tissue assay dexamethasone displayed significant activity, whereas the blueberry extract was inactive. For the abdominal constriction test, inhibitions of 49.0, 54.5, 53.5%, respectively, were observed for the crude extract, and 61.4% for indometacin. In the formalin test, the crude extract (200 and 300 mg kg (-1)) and indometacin inhibited only the second phase by 36.2, 35.3 and 45.8%, respectively. Considering that the crude extract of blueberry displayed antinociceptive and anti-inflammatory activity, its consumption may be helpful for the treatment of inflammatory disorders.
JPP 2007, 59: 591–596
© 2007 The Authors
Received August 11, 2006
Accepted December 18, 2006
DOI 10.1211/jpp.59.4.0015
ISSN 0022-3573
Anti-inflammatory and antinociceptive properties
of blueberry extract (Vaccinium corymbosum)
Eliane Torri, Marivane Lemos, Vinícius Caliari, Cândida A. L. Kassuya,
Jairo K. Bastos and Sérgio F. Andrade
Blueberries are among the edible fruits that are recognized best for their potential health ben-
efits. The crude extract from Vaccinium corymbosum was assessed in anti-inflammatory and
antinociceptive models. The crude hydroalcoholic extract was administered orally at doses of
100, 200 or 300 mgkg
for all the assays. In the carrageenan test, the crude extract reduced rat
paw oedema by 9.8, 28.5 and 65.9%, respectively. For the histamine assay, the reductions of
oedema were 70.1, 71.7 and 81.9%, respectively. In the myeloperoxidase (MPO) assay,
300 mgkg
crude extract produced a significant inhibition of the MPO activity, at 6 h and 24 h
after injection of carrageenan, by 42.8 and 46.2%, respectively. With the granulomatous tissue
assay dexamethasone displayed significant activity, whereas the blueberry extract was inactive.
For the abdominal constriction test, inhibitions of 49.0, 54.5, 53.5%, respectively, were observed
for the crude extract, and 61.4% for indometacin. In the formalin test, the crude extract (200
and 300 mgkg
) and indometacin inhibited only the second phase by 36.2, 35.3 and 45.8%,
respectively. Considering that the crude extract of blueberry displayed antinociceptive and anti-
inflammatory activity, its consumption may be helpful for the treatment of inflammatory
Blueberries are among the edible fruits that are recognized best for their potential health
benefits. Many of the health-promoting properties are thought to be attributable to their
main group of bioactive compounds, which belong to the proanthocyanidin and anthocy-
anin classes (Faria et al 2005). In this regard, the chemical profile of plants belonging to
the genus Vaccinium could be characterized mainly by the presence of anthocyanin
compounds in the fruits, which are called blueberry due to the high content of these com-
pounds. As a result, there has been growing interest in the anthocyanin content of some
Vaccinium species with regards to its pharmacological properties, particularly the effects
on blood vessels, its use in ophthalmology, and the potential inhibition of HIV (Cabrita &
Andersen 1999).
Anthocyanins are plant pigments responsible for the orange, red and blue colours of
fruits, flowers, vegetables and other storage tissues in plants (Strack & Wray 1993).
These compounds are involved in a wide range of biological activity (Kong et al 2003),
including antioxidant (Mazza et al 2002), anti-inflammatory (Youdim et al 2002) and
anticarcinogenic (Katsube et al 2003). Moreover, anthocyanins may also display neuro-
protective action and the ability to reduce the risk of coronary heart disease (Renaud &
De Lorgeril 1992) through vasoprotective activity, effects on arterial vasomotion, and
inhibition of platelet aggregation (Colanttuoni et al 1991). Anthocyanins from
Vaccinium are used to enhance vision and to increase capillary resistance (Peterson &
Dwyer 1998).
In the literature there are many compounds of plant origin reported as possessing import-
ant biological activity, such as anti-inflammatory (Namiki 1990). The purpose of this study
was to investigate the anti-inflammatory and antinociceptive effects of the hydroalcoholic
extract of blueberry (Vaccinium corymbosum).
Núcleo de Ciência e Tecnologia,
Área de Ciências Biológicas e da
Saúde, Universidade do Oeste de
Santa Catarina, Campus de
Videira, Rua Paese, 198,
Bairro das Torres, Videira-SC,
89560-000, SC, Brazil
Eliane Torri, Marivane Lemos,
Cândida A. L. Kassuya,
Sérgio F. Andrade
Programa de Mestrado em
Ciências Farmacêuticas,
Núcleo de Investigações
(NIQFAR), Universidade do Vale
do Itajaí – UNIVALI, Itajaí,
Santa Catarina, Brazil
Sérgio F. Andrade
Estação Experimental de Videira,
Empresa de Pesquisa
Agropecuária e Extensão Rural
do Estado de Santa Catarina
(EPAGRI) Videira, SC, Brazil
Vinícus Caliari
Faculdade de Ciências
Farmacêuticas de Ribeirão Preto,
Universidade de São Paulo,
Ribeirão Preto, SP, Brazil
Jairo K. Bastos
Correspondence: S. F. Andrade,
Núcleo de Ciência e Tecnologia,
Área de Ciências Biológicas e da
Saúde, Universidade do Oeste de
Santa Catarina, Campus de
Videira, Rua Paese, 198, Bairro
das Torres, Videira-SC,
89560-000, SC, Brazil. E-mail: Page 591 Tuesday, March 6, 2007 10:21 AM
592 Eliane Torri etal
Plant material
Berries of the blueberry (V. corymbosum) were collected
in the experimental field of “Pesquisa Agropecuária e
Extensão Rural” company of the State of Santa Catarina
(EPAGRI), Videira, in 2005. All berries were picked at the
commercially ripe stage. The berries were selected by
removing damaged, diseased, pest-infested fruits, stems
and leaves. They were maintained in polyethylene bags
at 20°C until extract preparation. Before extraction, fro-
zen berries were crushed by using a food processor. The
crushed berries (1.0 kg) were macerated with aqueous eth-
anol 70% (v/v) at room temperature for seven days. The
crude extract was obtained by filtration, followed by con-
centration under reduced pressure, yielding 138.0 g
(13.8%, w/w).
Phytochemical studies carried out with Vaccinium ber-
ries have demonstrated the presence mainly of phenolic
acid compounds, such as: gentisic, gallic, o-pyrocatechuic,
protocatechuic, salicylic, syringic, vanillic, veratric, caffeic,
m-coumaric, o-coumaric, p-coumaric, 3,4-dimethoxycin-
namic, ferulic, hydroxycaffeic, sinapic, and p-hydroxyphenil-
acetic. In addition many anthocyanin compounds have been
identified, such as: delphinidin-3-galactose, delphinidin-
3-glucose, delphinidin-3-arabinose, cyanidin-3-galactose,
cyanidin-3-glucose, petunidin-3-galactose, cyanidin-3-
arabinose, petunidin-3-glucose, peonidin-3-galactose, petuni-
din-3-arabinose, peonidin-3-glucose, malvidin-3-galactose,
peonidin-3-arabinose, malvidin-3-glucose and malvidin-
3-arabinose (Blumenthal et al 2000; Faria et al 2005;
Zadernowski et al 2005).
Quantification of total phenolics and
The concentration of phenolic compounds in the extract was
determined by the Folin-Ciocalteu colorimetric method
(Singleton & Rossi 1965). Analyses were carried out in tripli-
cate, and the quantification was calculated from a calibration
curve obtained with catequin. Total phenolics were expressed
as catequin equivalents (mg (g extract)
Anthocyanidins were quantified using a Varian Pro Star
HPLC system equipped with a type VA CP29257 C18 col-
umn (250 mm × 4.6 mm i.d., 5 m m, VARIAN), Pro Star 400
auto sampler and detector Pro Star 310 UV-vis at 524 nm.
The mobile phase consisted of water:acetonitrile (85:15, v/v),
acidified (pH 2.2) with phosphoric acid. The elution was
undertaken using an isocratic mode at a flow-rate of
1.2 mLmin
. Under analytical conditions standard samples
were injected (10 m L; 5, 15 and 25 mg L
) obtaining standard
curves with three levels, 10% of tolerance and r
coefficient = 0.999 for malvidin, cyanidin and delphinidin.
Detection was performed at 280 nm. Samples of extract were
filtered through a 0.45-m m syringe filter before injection of a
10-m L sample into the HPLC system. The concentrations of
malvidin, cyanidin and delphinidin were determined and
expressed as mg g
Male Wistar rats (200–250 g) and male Swiss mice (40–45 g)
were provided by the Central Animal House of University of
West of Santa Catarina (UNOESC). The animals were
housed in groups of five in standard cages at room tempera-
ture (25 ± 3°C), under a 12-h dark/light cycle, the real period
of light, with food and water freely available. Twelve hours
before the experiments animals were transferred to the labo-
ratory and were maintained only with water freely available.
The experiments were authorized by the Ethical Committee
for Animal Care of University of West of Santa Catarina,
Brazil, in accordance with the Federal Government legisla-
tion on animal care. All efforts were made to minimize
animal suffering and to reduce the number of animals used.
Carrageenan-induced rat paw oedema
The method used was described by Winter et al (1962).
Groups of rats (n = 6) were treated orally with blueberry crude
extract dissolved in water (100, 200 or 300 mg kg
indometacin (10 mg kg
) or vehicle (water), 30 min before
the injection of the stimulus (carrageenan 1000 m g/paw) into
the right hind paw plantar surface. Sterile saline solution
(0.9%, 0.1 mL) was injected into the left paw as control refer-
ence for the tested paw. The foot volumes of the animals were
determined by a plestimographic method described by
Ferreira (1979). Foot volume was measured before and at
hourly intervals for 3 h after the injection of the inflammatory
stimulus into the right hind paw plantar surface. The inhibi-
tion of inflammation was calculated by measuring the volume
difference between the right and left paws in comparison with
the control group.
Histamine-induced rat paw oedema
The animals were treated in a manner similar to that of the
carrageenan-induced paw oedema protocol. However, the
paw oedema was induced by subplantar injection of hista-
mine (50 m g/paw), and the oedema was measured as men-
tioned earlier at hourly intervals for 2 h. A positive control
group was treated with hydroxyzine (10 mg kg
, p.o.).
Myeloperoxidase activity
The anti-inflammatory activity of V. corymbosum was inves-
tigated by evaluating neutrophil infiltration indirectly by
measuring myeloperoxidase (MPO) activity, which was
determined at 6- and 24-h time points (De Young et al 1989).
The crude extract (300 mg kg
) and vehicle (water) were
administered to the animals as described above. Briefly, 6
and 24 h after carrageenan injection, the subcutaneous tissue
of the injected paws was removed and homogenized in 5%
(w/v) 80 m
M phosphate buffer, pH 5.4, containing 0.5% hexa-
decyltrimethylammonium bromide. The homogenate was
centrifuged at 12 000 g and 4°C for 20 min. Samples (30 mL)
of each supernatant were mixed with 100 mL phosphate buffer
80 m
M, 85 m L phosphate buffer 0.22 M and 15 m L 0.017%
in 96-well plates. The reaction was triggered with 20 mL
tetramethylbenzidine (dissolved in dimethylformamide). The
Materials and Methods Page 592 Tuesday, March 6, 2007 10:21 AM
Anti-inflammatory effects of Vaccinium corymbosum 593
plate was kept at 37°C for 3 min. It was then placed on ice,
and the reaction was stopped by adding 30 m L sodium acetate
M, pH 3.0. The enzymatic activity was determined by
measuring the optical density at 630 nm, and it was expressed
as mOD (mg protein)
Granulomatous tissue induction
This assay was described by Niemegeers (1975). Pellets
weighing approximately 40 mg each were made with 5 mm of
dental cotton tampons and implanted into rats (n = 6). The
pellets were sterilized and impregnated with 0.4 mL ampicil-
lin water solution at the moment of implantation. Animals
were anaesthetized with thiopental sodium, and four pellets
were subcutaneously introduced through an abdominal skin
incision. Each group was treated orally, daily for six consec-
utive days, with crude extract (300 mg kg
), dexamethasone
(0.2 mg kg
) or vehicle (0.5 mL water). On the seventh day,
the animals were killed, the pellets were dissected out, and
the wet weights were immediately determined. After that,
granulomas were dried at 60°C overnight to determine the
dried weight. The difference between the initial and final
weight was considered as the weight of the granulomatous
tissue produced.
Abdominal constriction test
The abdominal constriction test was carried out as described
by Koster et al (1959). Groups of mice (n = 8) were treated
with crude extract (100, 200 or 300 mg kg
), indometacin
(10 mg kg
) or vehicle (0.5 mL water), orally. The abdominal
constrictions were induced by an intraperitoneal injection of
0.6% acetic acid solution (0.25 mL/animal), 30 min after the
treatment. The number of abdominal constrictions was
counted starting at 5 min after acetic acid injection for a
period of 20 min. Data represented the average of the total
abdominal constrictions observed.
Formalin test
The test was carried out as described by Hunskaar & Hole
(1987). Animals were injected subcutaneously with 20 m L
formalin into the dorsal hind paw. Groups of mice (n = 8)
were treated with crude extract (100, 200 or 300 mg kg
indometacin (10 mg kg
) or vehicle (0.5 mL water), orally,
30 min before formalin injection. The time spent by mice
licking the injected paw was recorded. On the basis of the
response pattern described by Tjolsen et al (1992), two dis-
tinct periods of intensive licking activity were identified and
scored separately. The first period, early phase, was recorded
for the first 5 min, after the injection of formalin, and the
second period, late phase, was recorded between 20 and 30 min
after the injection of the stimulus.
Statistical analysis
All the data were analysed for n 5. Data were reported as
mean ± s.e.m., and it was analysed statistically by analysis of
variance, followed by Dunnett’s test (Sokal & Rohlf 1995).
Results with P < 0.05 were considered significant.
The total concentration of phenolic compounds in the extract
was 5.66 ± 0.01 mg catechuin g
. Malvidin, cyanidin and
delphinidin were present in the extract at concentrations of
0.131 ± 0.06, 0.099 ± 0.05 and 0.063 ± 0.06 mg g
, respec-
tively (Figure 1).
Figures 2 and 3 show the effect of V. corymbosum crude
extract on the carrageenan- and histamine-induced rat paw
oedema, respectively. In the carrageenan protocol, the crude
extract reduced the paw oedema, measured 3 h after carra-
geenan injection, which corresponded to the peak of oedema
formation, by 9.8, 28.5 and 65.9%, at doses of 100, 200 and
300 mgkg
, respectively. For the group treated with
indometacin, a 63.8% reduction of oedema was observed.
These results were significant (P < 0.05) only for the groups
treated with 300 mg kg
crude extract or indometacin
(Figure 2). In the histamine protocol, a significant reduction
of the oedema formation (P < 0.05) was observed at 1 h,
which corresponded with the peak of oedema. For the groups
treated with crude extract at 100, 200 or 300 mgkg
, inhibi-
tions of 70.1, 71.7 and 81.9%, respectively, were observed.
The positive control, hydroxyzine (10 mgkg
), reduced the
oedema by 68.6% (Figure 3).
With the myeloperoxidase assay, the crude extract of
V. corymbosum (300 mgkg
) produced a significant inhibi-
tion of MPO activity in the treated animals, at 6 and 24 h after
injection of carrageenan by 42.8 and 46.2%, respectively
(Figure 4).
In the granulomatous tissue induction assay, on one hand
dexamethasone displayed significant inhibition (P < 0.05) in
both wet and dry granuloma weights, and on the other hand,
the crude extract neither diminished the wet nor the dry gran-
uloma weights (Table 1).
Figure 1 HPLC chromatographic profile of the blueberry hydroalco-
holic crude extract, using a C18 column and an isocratic mobile phase
consisting of water:acetonitrile (85:15, v/v), acidified (pH 2.2) with
phosphoric acid. 1, Cyanidin; 2, delphinidin; 3, malvidin.
5 1015202530
Time (min)
–27 Page 593 Tuesday, March 6, 2007 10:21 AM
594 Eliane Torri etal
In the abdominal constriction test, crude extract (100, 200
or 300 mgkg
) and indometacin were significantly effective
in inhibiting the abdominal constrictions in mice, compared
with the control group. Abdominal constriction inhibitions
were 49.0, 54.5, 53.5 and 61.4% for the groups treated with
100, 200 or 300 mgkg
crude extract and indometacin,
respectively (Table 2).
In the formalin test, the administration of crude extract and
indometacin inhibited only the second phase of the test,
which corresponded to inflammatory pain. The results were
significant for the groups treated with 200 or 300 mgkg
crude extract and indometacin, displaying 36.2, 35.3 and
45.8% reduction in licking time, respectively (Table 3).
This study demonstrated that the blueberry (V. corymbosum)
hydroalcoholic extract could play a significant role in the
inhibition of pain and inflammatory processes. Inflammation
is an essential protective process to preserve the integrity of
organisms against physical, chemical, and infective aggres-
sion (Walport & Duff 1993). However, the inflammatory
response frequently leads to the damage of normal tissue
(Majno & Joris 1996).
Among the protocols used to screen new anti-inflammatory
agents, carrageenan-induced rat paw oedema is the most
widely used primary test (Winter et al 1962). The development
of oedema in rat paw, after the injection of carrageenan, has
been described as a biphasic event (Brito & Antonio 1998).
The initial phase, observed during the first hour, has been
attributed to the release of histamine and serotonin, and the
second phase to the release of prostaglandins (Crunkhorn &
Meacock 1971). Therefore, it could be inferred that the signi-
ficant activity observed in the suppression of the second phase
may be explained by an inhibition of cyclooxygenase (Badilla
et al 2003). Moreover, indometacin, the usual and non-selective
inhibitor of cyclooxygenase, inhibits the second phase of the
carrageenan-induced oedema (Tamura et al 2002). In addition,
histamine is a basic amine related to the inflammatory and
allergic process causing, among several effects, vasodilatation
and an increase in vascular permeability (Rang et al 2001). The
higher dose of the V. corymbosum extract (300 mgkg
) inhib-
ited carrageenan and histamine oedema models, indicating
anti-inflammatory and antihistaminic effects.
In the myeloperoxidase assay, a significant reduction in
MPO activity for the group treated with blueberry extract was
observed. The onset of the carrageenan oedema has been
linked to neutrophil infiltration, release of other neutrophil-
derived mediators and production of neutrophil-derived free
radicals (Cuzzocrea et al 1999). MPO, a haeme protein, is an
enzyme present in neutrophils and at a much lower concentra-
tion in monocytes and macrophages. It is a critical enzyme for
the generation of hypochlorous acid and other toxic oxygen
products. It is well known that the level of MPO activity is
directly proportional to the neutrophil concentration in the
inflamed tissue (Bradley et al 1982). Hence, the measurement
of the enzyme activity has been considered a quantitative and
sensitive tool to evaluate the chemotaxis and neutrophil
infiltration in the inflammatory process (Smith 1994).
Figure 2 Effect of the administration of blueberry crude extract (100,
200 and 300 mg kg
) and indometacin (10 mg kg
) on carrageenan-
induced rat paw oedema. The paw oedema volume was taken 3 h afte
carrageenan injection. Each column represents the mean ± s.e.m., n = 6.
*P < 0.05, analysis of variance followed by Dunnett’s test.
100 mg kg
200 mg kg
300 mg kg
Oedema (mL)
Figure 3 Effect of the administration of blueberry crude extract (100,
200 and 300 mg kg
) and hydroxyzine (10 mg kg
) on histamine-
induced rat paw oedema. The paw oedema volume was taken 1 h afte
histamine injection. Each column represents the mean ± s.e.m., n = 6.
*P < 0.05, analysis of variance followed by Dunnett’s test.
100 mg kg
200 mg kg
300 mg kg
Oedema (mL)
Figure 4 Effect of the administration of blueberry crude extract
(300 mg kg
) on myeloperoxidase activity. The myeloperoxidase activ-
ity was measured at 0, 6 and 24 h after carrageenan injection. Each col-
umn represents the mean ± s.e.m., n = 6. *P < 0.05, analysis of variance
followed by Dunnett’s test.
6 h
24 h
300 mg kg
MPO activity
(mOD (mg protein)
Discussion Page 594 Tuesday, March 6, 2007 10:21 AM
Anti-inflammatory effects of Vaccinium corymbosum 595
Nevertheless, it is also known that inflammation sites
present high concentrations of free radicals and oxidants,
which play an important role in different inflammation proc-
esses. Therefore, antioxidant compounds may be helpful to
prevent this process (Salvemini et al 1996). In this regard,
antioxidant activity has been described for several flavonoids
and anthocyanins (Peterson & Dwyer 1998; Moyer et al
2002), which are present in high concentrations in the fruits
of Vaccinium species, such as malvidin, cyanidin, delphini-
din, astragalin, hyperoside, isoquercitrin and quercitrin
(Blumenthal et al 2000). Hence, these compounds may play
an important role in the anti-inflammatory activity of blue-
berry extract, not only because of its actions on inflammatory
chemotatic mediators, but also its reported antioxidant activ-
ity. It is known that the inflammatory granuloma is a typical
response of a chronic inflammatory process. It has been
established that the dry weight of the pellets is well correlated
with granulomatous tissue (Olajide et al 2000). Anti-
inflammatory steroidal drugs show higher activity in this
model (Swingle & Shideman 1972). However, the crude
extract of V. corymbosum was not able to significantly inhibit
this process. Regarding the antinociceptive activity, the intra-
peritoneal administration of acetic acid irritates serous mem-
branes, provoking a stereotypical behaviour in mice
characterized by abdominal contractions, movements of the
body as a whole, twisting of dorsoabdominal muscles, and a
reduction in motor activity and coordination (Bars et al 2001).
Acetic acid causes algesia by liberating endogenous sub-
stances that excite pain nerve endings, and it is a sensitive
method for screening peripheral and central analgesic agents
(Collier et al 1968). The administration of crude extract of
V. corymbosum reduced the number of mouse abdominal
constrictions after acetic acid administration, indicating anal-
gesic activity for this extract at the assayed doses. The
obtained results for the abdominal constriction test alone did
not allow the determination as to whether the antinociceptive
effect was either central or peripheral. Thus, to clear the mode
of the inhibitory effects of this extract on the nociceptive
responses, the formalin test was used. In the formalin test,
there was a distinct biphasic response. The initial pain, early
phase, was explained as a direct stimulation of nociceptors.
The late phase is thought to be secondary to the inflammatory
reactions (Hunskaar & Hole 1987). Therefore, the test can be
used to clarify the possible mechanism of antinociceptive
effect of a proposed analgesic (Tjolsen et al 1992). On one
hand, centrally acting drugs, such as opioids, inhibit both
phases equally (Shibata et al 1989), but on the other hand,
peripherally-acting drugs such as aspirin, indometacin and
dexamethasone inhibit only the late phase. The crude extract
of V. corymbosum inhibited only the second phase of the for-
malin test, suggesting peripheral analgesic activity.
The crude extract of berries from V. corymbosum displayed
antinociceptive and anti-inflammatory activity. Considering
that blueberries are rich in phenolic acids, flavonoids and
anthocyanins, and that there are several works reporting the
anti-inflammatory and antinociceptive activity for com-
pounds belonging to these classes, it is suggested that the
reported activity might have been, at least in part, due to these
Table 1 Effect of the administration of blueberry crude extract and dexamethasone on cotton pellet-induced granuloma weights
The results are expressed as mean ± s.e.m. (n = 6). *P < 0.05, analysis of variance followed by Dunnett’s test.
Groups Dose
(mg kg
Mean weight
of wet granuloma (mg)
Inhibition (%) Mean weight
of dry granuloma (mg)
Inhibition (%)
Control – 714.2 ± 27.8 133.2 ± 5.0 –
Crude extract 300 570.2 ± 16.8 20.1 112.4 ± 3.6 15.6
Dexamethasone 0.2 392.0 ± 6.5* 45.1 79.6 ± 2.1* 40.2
Table 2 Effect of the administration of blueberry crude extract and
indometacin on acetic acid-induced abdominal constrictions
The results are expressed as mean ± s.e.m., n = 8. *P < 0.05, analysis of
variance followed by Dunnett’s test.
Groups Dose
(mg kg
Number of
(mean ± s.e.m.)
Control – 92.7 ± 5.7 –
Crude extract 100 49.8 ± 2.7* 49.0
200 42.1 ± 2.3* 54.5
300 43.1 ± 2.5* 53.5
Indometacin 10 35.7 ± 4.7* 61.4
Table 3 Effect of the administration of blueberry crude extract and
indometacin on formalin-induced nociception
The results are expressed as mean ± s.e.m., n = 8. *P < 0.05, analysis
followed by Dunnett’s test.
Groups Dose
(mg kg
Licking time (s)
(mean ± s.e.m.)
Early phase
(0–5 min)
Late phase
(20–30 min)
Control – 66.5 ± 3.2 114.2 ± 4.5 –
Crude extract 100 55.3 ± 4.9 93.0 ± 6.1 16.8 18.5
200 63.0 ± 6.6 72.8 ± 8.9* 5.2 36.2
300 72.3 ± 9.3 73.8 ± 2.5* 8.7 35.3
Indometacin 10 66.8 ± 7.5 61.8 ± 9.1* 0.4 45.8 Page 595 Tuesday, March 6, 2007 10:21 AM
596 Eliane Torri etal
compounds. Moreover, considering that blueberries are edi-
ble fruits, their consumption may be helpful for the treatment
of inflammatory disorders. In addition, this work has corrobo-
rated the traditional indication of different species of Vaccinium
to treat inflammatory conditions, which may contribute to the
understanding of the role of this edible fruit in promoting
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... Similarly, in different anti-inflammatory tests, Vaccinium exhibited high anti-inflammatory activity [11]. High concentrations of anthocyanins (such as cyanidin, delphinidin and malvidin) and flavonoids (such as astragalin, hyperoside, isoquercitrin, and quercitrin) appear to be related to the anti-inflammatory and antioxidant activities ascribed to these berries [94,95]. Considering that berries of Vaccinium are edible, their consumption may be helpful for the treatment of inflammatory illnesses. ...
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The genus Vaccinium L. (Ericaceae) includes more than 450 species, which mainly grow in cooler areas of the northern hemisphere. Vaccinium species have been used in traditional medicine of different cultures and the berries are widely consumed as food. Indeed, Vaccinium supplement-based herbal medicine and functional food, mainly from V. myrtillus and V. macrocarpon, are used in Europe and North America. Biological studies support traditional uses since, for many Vaccinium components, important biological functions have been described, including antioxidant, antitumor, anti-inflammatory, antidiabetic and endothelium protective activities. Vaccinium components, such as polyphenols, anthocyanins and flavonoids, are widely recognized as modulators of cellular pathways involved in pathological conditions, thus indicating that Vaccinium may be an important source of bioactive molecules. This review aims to better describe the bioactivity of Vaccinium species, focusing on anti-inflammatory and endothelial protective cellular pathways, modulated by their components, to better understand their importance for public health.
... Total polyphenols concentrations were: MA: 1906.5 ± 73.2; CA: 1344.2 ± 10.5; BL: 1229.6 ± 20.9 mg GAE/100 g of dry weight. The levels observed were lower than the ones observed in previous works [32][33][34]. This difference could be due to the use of a non-acidified extraction. ...
BACKGROUND: Obesity occurs due to a positive energy imbalance, leading to the expansion of adipose tissue. This phenomenon triggers a chronic low-grade inflammatory state, which is associated with comorbidities development. It is, therefore, of great interest to investigate new counteracting nutritional strategies. In this regard, polyphenol-rich Chilean native fruits, Aristotelia chilensis (Maqui) and Berberis microphylla (Calafate), and also the non-Chilean Vaccinium corymbosum (Blueberry), have been associated with antioxidant and anti-inflammatory features. OBJECTIVE: To evaluate Maqui, Calafate, and Blueberry aqueous extracts treatments on the pathogenic response of human activated macrophages and visceral adipocytes. METHODS: THP-1 monocyte human cell line and differentiated human visceral preadipocytes were activated (with lipopolysaccharide and TNF-␣, for 48 and 24 h, respectively), and treated with the aqueous extracts. Inflammation and oxidative stress markers were assessed. A U T H O R C O P Y 532 A. Ovalle-Marin et al. / Berries extracts block inflammation in vitro RESULTS: Lower NO and IL-6 secretion, and inhibited apoptosis in activated macrophages, were observed. Also, decreased gene expression of MCP-1 and secretion of IL-6, inhibited apoptosis, and increased levels of GSH in activated adipocytes were detected. CONCLUSIONS: Maqui, Calafate, and Blueberry extracts showed anti-inflammatory and antioxidant responses in human macrophages and adipocytes.
... The many beneficial properties of blueberry extract, such as its anti-inflammatory properties, are thought to be attributable to their group of bioactive compounds. These bioactive compounds belong to the proanthocyanidin and anthocyanin classes [11]. Another component of NT-020 is green tea extract, which is composed of catechins (most abundant is epigallocatechin gallate or EGCG), theanine, and caffeine. ...
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Purpose: Evaluate the effectiveness of a commercially available nutraceutical product on levels of exercise-related energy, fatigue, and exertion before and after 4 weeks of supplementation. Primary ingredients within the product include a proprietary blend of blueberry extract, green tea extract, L-carnosine, Vitamin D3 (NT-020; 920mg), and rhodiola rosea, a plant that purports to boost energy and hold antioxidant and anti-inflammatory properties. Methods: Twenty-seven participants (13 females, 14 males, mean BMI = 23, mean age ± SD = 24 + 6 years) completed a baseline assessment of aerobic fitness (mean VO2 peak = 40 mL x kg-1 x min-1) before being randomized into a placebo or supplement group for four weeks. All participants were involved in regular physical activity three or more days per week. Assessment of energy, fatigue, and perceived exertion responses during and immediately following moderately intense cycle ergometry exercise was conducted before and after the 4-week ingestion period during which participants were instructed to maintain existing exercise activities. Results: Data were analyzed by way of repeated measures ANOVA and dependent t-tests to determine the presence of significant differences across time and between the supplement and placebo conditions. Participants receiving the supplement reported: greater levels of energy and lower levels of fatigue during the initial moments after completing the exercise trial (p < 0.05), greater levels of energy at the midpoint of the exercise trial (p < 0.05) but not at the end of the exercise session (p > 0.05), and lower perceived exertion at four of the six measurement points during exercise (p < 0.05). No differences were observed from pre to post intervention within the placebo condition (p > 0.05). Conclusions: Findings indicate that a commercially available supplement marketed to boost energy and reduce fatigue can deliver the purported benefits at least in part. Related findings that supplementation for a 4-week period can allow for equal work at a lower rating of perceived exertion provides further, though limited support that this product may have efficacy. Keywords: fatigue; perceived exertion; energy; rhodiola rosea; NT-020
... [14][15][16] The effect of plants that are used for their analgesic effect in traditional medicine may be, therefore, originating from ferulic acid. For instance, blueberries (Vaccinium corymbosum), which were shown to exhibit analgesic effect 27 , were reported to contain 66.5-68.3% of ferulic acid. 28 However, there are limited number of studies conducted on the peripheral analgesic effect mechanism of ferulic acid. ...
... Their protective effects may reduce the risk of coronary artery disease [58]. They have been shown to inhibit platelet aggregation [59]. ...
Kokilaksha Kashayam is a polyherbal Ayurvedic formulation prescribed for treating pain and inflammation. The scientific validation of the claimed pharmacological activities of this formulation has not been established yet. In the present work, the anti-inflammatory and Analgesic activity of freshly prepared in house Kokilaksha Kashayam as well as its marketed formulation were evaluated on experimental animals. The Anti-inflammatory activity was evaluated by carrageenan induces paw edema method, while the analgesic activity was evaluated by means of Eddy’s hot plate, tail-flick and formaldehyde induced tail flick methods. The results indicated that both in-house as well as marketed Kokilaksha Kashayam possess profound anti-inflammatory as well as analgesic property.
Anthocyanins possess strong antioxidant and anti-inflammatory physiological activities. This chapter summarizes the antioxidant and anti-inflammatory activities of anthocyanins from different food sources and discusses the mechanism of the above two activities. The antiultraviolet activity of anthocyanins and the physiological activity of improving osteoporosis are also introduced.
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Background Caffeic acid O -methyltransferases ( COMTs ) play an important role in the diversification of natural products, especially in the phenylalanine metabolic pathway of plant. The content of COMT genes in blueberry and relationship between their expression patterns and the lignin content during fruit development have not clearly investigated by now. Results Ninety-two VcCOMTs were identified in Vaccinium corymbosum . According to phylogenetic analyses, the 92 VcCOMTs were divided into 2 groups. The gene structure and conserved motifs within groups were similar which supported the reliability of the phylogenetic structure groupings. Dispersed duplication (DSD) and whole-genome duplication (WGD) were determined to be the major forces in VcCOMTs evolution. The results showed that the results of qRT-PCR and lignin content for 22 VcCOMTs , VcCOMT40 and VcCOMT92 were related to lignin content at different stages of fruit development of blueberry. Conclusion We identified COMT gene family in blueberry, and performed comparative analyses of the phylogenetic relationships in the 15 species of land plant, and gene duplication patterns of COMT genes in 5 of the 15 species. We found 2 VcCOMTs were highly expressed and their relative contents were similar to the variation trend of lignin content during the development of blueberry fruit. These results provide a clue for further study on the roles of VcCOMTs in the development of blueberry fruit and could promisingly be foundations for breeding blueberry clutivals with higher fruit firmness and longer shelf life.
Ageing is an unavoidable, universal, biological phenomenon affecting all organisms, which involves variable declines of individuals motor and memory capabilities. This study aimed to investigate the potential ameliorating effects of curcumin C3 complex, Astragalus membranaceus and blueberry on certain age-related biochemical alterations in rat liver. Four groups of rats, aged 12 months-old, were used. The first group; aged control group in which rats were left without any treatment until the age of 17 months. The other three groups received daily by oral gavage for 5 months the following supplements; curcumin C3 complex (110 mg/kg), Astragalus membranaceus (100 mg/kg) and blueberry (100 mg/kg) respectively. Additionally, a fifth group of rats, aged 5 months-old, was used as an adult control group. Our supplements alleviated ageing-induced redox state imbalance and inflammation as evidenced by reduction of hepatic thiobarbituric acid reactive substances and 8-hydroxydeoxyguanosine levels, restoration of total antioxidant capacity and nitric oxide contents, and lessening of lipofuscin deposition. All supplements decreased hepatic interlukin-6 gene expression and serum levels. Notably, Astragalus membranaceus and blueberry upregulated hepatic telomerase reverse transcriptase gene expression and increased telomere length. Our findings recommend the use of these natural hepatoprotective supplements for the elderly to promote healthy ageing and minimize the risk of age-related liver diseases.
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Fruits come in a wide variety of colors, shapes, and flavors. This chapter will cover selected fruits that are known to be healthy and highly nutritious. These fruits were chosen due to their common usage and availability. Since it is not possible to cover all health benefits or essential nutrients and important phytochemicals of the fruit composition, this chapter will focus on the key valuable constituents and their potential health effects.
We examined the hypothesis that myeloperoxidase (MPO), a plentiful constituent of neutrophils, might serve as a marker for tissue neutrophil content. To completely extract MPO from either neutrophils or skin, hexadecyltrimethylammonium bromide (HTAB) was used to solubilize the enzyme. With this detergent treatment, 97.8 +/- 0.2% of total recoverable MPO was extracted from neutrophils with a single HTAB treatment; 93.1 +/- 1.0% was solubilized with a single treatment of skin. Neutrophil MPO was directly related to neutrophil number; with the dianisidine-H2O2 assay as few as 10(4) neutrophils could be detected. The background level of MPO within uninflamed tissue was 0.385 +/- 0.018 units per gram of tissue, equivalent to only 7.64 +/- 0.36 X 10(5) neutrophils. In experimental staphylococcal infection, skin specimens contained 34.8 +/- 3.8 units MPO per gram, equivalent to 8.55 +/- 0.93 X 10(7) neutrophils. These studies demonstrate that MPO can be used as a marker for skin neutrophil content: it is recoverable from skin in soluble form, and is directly related to neutrophil number. Further, normal skin possesses a low background of MPO compared to that of inflamed skin.
The role of arachidonic metabolites in the control of immunity in inflammation is reviewed with particular emphasis on the most recent observations and possible interactions between these products and other mediators. Continuing studies on the nature of the lipoxygenase involved in leukotriene synthesis will be discussed as will the nature of the enzymatic activity involved in the transfer of sulphur in leukotriene C, D and E. The most important known lipid mediators of inflammation are either metabolites of long chain fatty acids, particularly arachidonic acid, or 1-alkyl- 2-acetyl derivatives of phosphatidylcholine (platelet activating factor, PAF). This chapter will review the mediators derived from arachidonic acid with emphasis on the most recent findings, particularly in regard to the lipoxygenase products, which is a very active area. [1–6].
In most countries, high intake of saturated fat is positively related to high mortality from coronary heart disease (CHD). However, the situation in France is paradoxical in that there is high intake of saturated fat but low mortality from CHD. This paradox may be attributable in part to high wine consumption. Epidemiological studies indicate that consumption of alcohol at the level of intake in France (20-30 g per day) can reduce risk of CHD by at least 40%. Alcohol is believed to protect from CHD by preventing atherosclerosis through the action of high-density-lipoprotein cholesterol, but serum concentrations of this factor are no higher in France than in other countries. Re-examination of previous results suggests that, in the main, moderate alcohol intake does not prevent CHD through an effect on atherosclerosis, but rather through a haemostatic mechanism. Data from Caerphilly, Wales, show that platelet aggregation, which is related to CHD, is inhibited significantly by alcohol at levels of intake associated with reduced risk of CHD. Inhibition of platelet reactivity by wine (alcohol) may be one explanation for protection from CHD in France, since pilot studies have shown that platelet reactivity is lower in France than in Scotland.
Twenty anthocyanins were isolated from the extract of the edible blue berries of Vaccinium padifolium (Uveira) by a combination of chromatographic techniques. Mainly on the basis of nuclear magnetic resonance techniques (1H, COSY, TOCSY, HSQC, HMBC, SEFT) they were identified as the 3-O-β-glucopyranosides, 3-O-β-galactopyranosides, 3-O-β-arabinopyranosides and 3-O-sambubiosides (2″-O-β-xylopyranosyl-O-β-glucopyranosides) of delphinidin, cyanidin, petunidin, malvidin and peonidin. Anthocyanidin disaccharides have not been identified in the genus Vaccinium previously, and the 3-sambubiosides of peonidin, petunidin and malvidin are novel compounds.
Although antioxidant systems help control the level of reactive oxygen species they may be overwhelmed during periods of oxidative stress. Evidence suggests that oxidative stress components as well as inflammatory mediators may be involved in the pathogenesis of vascular disorders, where localized markers of oxidative damage have been found. In this regard we investigated the putative antioxidant and anti-inflammatory effects of blueberry and cranberry anthocyanins and hydroxycinnamic acids against H2O2 and TNFα induced damage to human microvascular endothelial cells. Polyphenols from both berries were able to localize into endothelial cells subsequently reducing endothelial cells vulnerability to increased oxidative stress at both the membrane and cytosol level. Furthermore, berry polyphenols also reduced TNFα induced up-regulation of various inflammatory mediators (IL-8, MCP-1 and ICAM-1) involved in the recruitment of leukocytes to sites of damage or inflammation along the endothelium. In conclusion, polyphenols isolated from both blueberry and cranberry were able to afford protection to endothelial cells against stressor induced up-regulation of oxidative and inflammatory insults. This may have beneficial actions against the initiation and development of vascular diseases and be a contributing factor in the reduction of age-related deficits in neurological impairments previously reported by us.
The role of peroxynitrite generated from nitric oxide and superoxide anion was investigated in a model of acute inflammation induced by the injection of carrageenan into the rat hind paw. Paw edema was inhibited 8 h following the administration of carrageenan by N-iminoethyl-l-lysine (3–30 mg/kg, n = 6) or aminoguanidine (30–300 mg/kg, n = 6), two selective inhibitors of inducible nitric oxide synthase and by recombinant human superoxide dismutase coupled to polyethyleneglycol (12 × 103 U/kg, n = 6, P < 0.001). Moreover, at the same time point following carrageenan administration, intense immunoreactive staining for nitrotyrosine (a marker of peroxynitrite formation) was detected. Our results suggest that the generation of nitric oxide, superoxide anion and peroxynitrite contributes to the edema observed in this acute model of inflammation.