Effect of Fish or Soybean Oil-Rich Diets on Bradykinin,Kallikrein, Nitric Oxide, Leptin, Corticosteroneand Macrophages in Carrageenan Stimulated Rats
Marta Wohlers,1 Roberta Araujo Navarro Xavier,1 Lila Missae Oyama,1
Eliane Beraldi Ribeiro,1 Claudia Maria Oller do Nascimento,1
Dulce Elena Casarini,2 and Vera Lucia Flor Silveira1,3
Abstract—We have previously demonstrated that both nY3 and nY6 polyunsaturated fatty acids
(PUFA)-rich diets decrease the acute inflammatory response partially explained by the high cor-
ticosterone basal levels. The present study aimed to determine the effect of hyperlipidic diets
(PUFA nY3 or nY6) on phagocytosis, hydrogen peroxide (H2O2) and nitric oxide (NO) release by
macrophages, bradykinin (BK) and NO release in the paw inflammatory perfusate and Kallikrein
(KK), corticosterone and leptin blood levels. Hyperlipidic diets decreased H2O2 release from
macrophages stimulated by carrageenan or phorbol-miristate-acetate (PMA), NO release from
macrophage stimulated by carrageenan, BK and NO release in the edema perfusate, KK plasma
levels and the increase of serum leptin after carrageenan stimulus. These data show that both fish
and soybean oil-rich diets promote similar alterations on inflammatory mediators of carrageenan
edema and a causal association with the anti-inflammatory effect of these diets.
KEY WORDS: polyunsaturated fatty acids; inflammation; corticosterone; bradykinin; nitric oxide; hydrogenperoxide; leptin.
INTRODUCTION
It has been demonstrated that dietary lipids are
incorporated into phospholipids cell membranes chang-
ing its fluidity, activity and release of intracellular
mediators [1, 2]. Many studies have shown that
alterations in the type and quantity of polyunsaturated
fatty acids (PUFA) from diet could modify immuno-
logical and inflammatory responses [3, 4].
Although the exact mechanisms responsible for
these effects are not fully elucidated, there have been
recent reports showing that PUFAs such as DHA
(docosahexaenoic acid, 22:6, nY3) and AA (arachidonic
acid, 20:4, nY6) induce the production of anti-inflam-
matory mediators, named docosatrienes, protectins,
lipoxins, and resolvins [5, 6]. Those eicosanoids have
been shown to play an important role during the
resolution phase of the inflammatory process and could
help explain the anti-inflammation induced by diets
enriched with those PUFAs [7].
Additionally, clinical studies have shown beneficial
effects of dietary supplementation with fish oil (rich in
nY3 PUFA), under chronic inflammatory conditions [8,
9] and these effects have been attributed to a decrease in
the generation of inflammatory mediators derived from
the AA, with the production of less potent inflammatory
mediators, after nY3 PUFA-rich diets treatment [9Y11].
0360-3997/06/0200-0081/0 # 2006 Springer Science+Business Media, Inc.
Inflammation, Vol. 29, Nos. 2Y3, April 2005 (# 2006)
DOI: 10.1007/s10753-006-9002-2
81
1 Physiology Department, Federal University of Sao PauloVEPM,
Sao Paulo, Brazil.2 Medicine Department, Federal University of Sao PauloVEPM, Sao
Paulo, Brazil.3 To whom correspondence should be addressed at Departamento de
Fisiologia, Disciplina de Fisiologia da Nutricao, Universidade
Federal de Sao PauloVEscola Paulista de Medicina, Rua Botucatu,
862-2- andar, Vila ClementinoVCEP, Sao Paulo 04023-060, Brazil.
E-mail: [email protected]
However, we previously observed that both fish
and soybean oil-rich diets were able to attenuate the
acute inflammatory process, which could be partially
attributed to increased corticosterone levels [12].
The inflammatory process is a response of the body
to injury, aimed at attacking the agent while limiting
and/or repairing tissue damage, thus helping the
reestablishment of homeostasis. The acute inflammatory
response is characterized by the development of
erythema, pain, and edema, as a result the local release
of inflammatory mediators, such as BK, NO, catechol-
amines, eicosanoids, and others [13, 14]. Macrophages
have an essential role on the inflammatory process and
dietary manipulations can change their fatty acids
composition, leading to alterations in the citotoxic
activity of these cells, such as phagocytosis and
production of reactive oxygen species [15].
The purpose of the present study was to examine
the effect of the chronic consumption of diets rich in
nY6 or nY3 PUFAs, on aspects of the acute inflamma-
tion process. For this, we evaluated macrophages
functionality and local as well as plasma release of
carrageenan edema mediators, as BK, KK, NO, and
glucocorticoids. Secondly, we aimed at verifying the
effect of these diets on plasma leptin, a hormone
released mainly from adipose tissue and involved in
the inflammatory process [16]. The association between
those mediators and the anti-inflammatory effect of the
lipid diets was examined.
MATERIAL AND METHODS
Animals and Treatment
The Committee on Experimental Research Ethics
from Federal University of Sao Paulo approved all
procedures involving animals.
Male Wistar rats (28Y30 days) weighing 65Y70 g
were supplied by the animal care facility of the Federal
University of Sao Paulo. They were housed under
controlled conditions of lighting (12:12 h lightYdark
cycle with lights on at 7:00 A.M.) and temperature (24 T1-C). During seven weeks, the rats had free access to
water and one of three types of diets: standard balanced
chow with 4% fat and 20% protein (Nuvilab) (control
group), nY6 PUFA-rich-diet prepared by adding 15% of
soybean oil to control diet (soybean group), or nY3
PUFA-rich diet prepared by adding 15% of fish oil
(Sigma) to the control diet (fish group). Twelve percent
of casein was added to the lipid diets to achieve 20% of
protein. Caloric density of diets was determined using an
adiabatic calorimeter IKA-C400. They were 17.4 KJ/g
for the standard chow and 20.5 KJ/g for both PUFA-rich
diets. Once prepared, the diets were kept frozen until
usage. The rats were provided with a fresh food cup
every day and a 24-h food intake measurement was
performed once a week. The fatty acids composition of
the diets was analyzed at the Nutrition Institute of
Federal University of Rio de Janeiro (Table 1).
Carrageenan-Induced Paw Edema
The animals were anesthetized with sodium pen-
tobarbital (40 mg/kg, i.p.). Anesthetic supplementation
(20 mg/Kg) was provided as required to keep a light
anesthesia level during the whole experiment. Carra-
geenan (0.1 mg) was injected in the right subplantar
region. Some animals were decapitated after 1, 2, 3, or
4 h later and trunk blood collected. Plasma corticoste-
rone levels were determined by fluorometry [17]. Serum
leptin was determined in the 1- and 3-h samples by
radioimmunoassay (Linco Research, Inc, USA).
In other carrageenan-stimulated animals, the tail
vein was punctured both 1 and 2 h after carrageenan for
Table 1. Fatty Acids Composition (g/100 g of diet) of Control, Fish,
and Soybean Diets
Fatty acid Control Fish Soybean
14:0 0.061 1.01 0.04
16:0 0.66 3.33 1.80
18:0 0.100 0.55 0.50
14:1 0.048 0.12 0.09
16:1 0.004 1.21 0.015
17:1 Y 0.10 Y18:1 nj9 0.80 2.02 3.43
24:1 nj9 0.002 0.05 Y18:2 nj6 2.11 1.71 8.08
18:3 nj6 0.008 0.015 0.04
18:3 nj3 0.13 0.09 0.81
20:3 nj3 Y 0.07 Y20:4 nj6 Y Y Y22:4 nj6 Y 0.06 Y22:5 nj3 Y 0.26 YEPA 0.002 1.44 0.011
DHA 0.01 1.58 0.004
Saturated 0.83 4.90 2.34
Monounsaturated 0.85 3.51 3.54
Polyunsaturated 2.26 5.22 8.94
nj6 Polyunsaturated 2.12 1.78 8.12
nj3 Polyunsaturated 0.14 3.44 0.82
Polyunsaturated: saturated 2.72 1.06 3.82
nj6 : nj3 15.14 0.52 9.90
82 Wohlers, Xavier, Oyama, Ribeiro, Nascimento, Casarini, and Silveira
determination of plasma kallikrein (KK) activity. This
was performed by measuring the production of p-nitro
anile (pNA) from the hydrolysis of the synthetic
substrate, S2302 [18].
For determination of local levels of nitric oxide
(NO) and bradykinin (BK), a coaxial perfusion of the
hind paw was performed (modified from [19]). Briefly,
the paw was cannulated before carrageenan injection
and perfused at 53 ml/min, with perfusion fluid (158
mM NaCl, 5.65 mM CaCl, 0.54 mM CaCl2, 2.90 mM
MgCl2, 178 mM NaHCO3, and 1.0 g/l glucose, main-
tained in bath at 36.5-C). Two one-hour paw perfusate
samples were collected, from 1 to 2 h and from 2 to 3
h after carrageenan, into ice-cold tubes. The samples
were centrifuged at 2.500 rpm, at 4-C, for 15 min and
the supernatant used for NO quantification. For BK
determination, the perfusate was collected into 80%
ethanol, centrifuged, and the supernatant evaporated in
gaseous nitrogen. The samples were stored at Y70-C.
NO levels were evaluated by nitrite (NO2Y) production,
using Griess reagent [20]. BK levels were determined
by reverse-phase HPLC [21].
The volume of the perfused paw was determined
by plethysmography (H. Basile, Italia), both before and
3 h after carrageenan administration.
Carrageenan-Induced Peritoneal Inflammation
The anesthetized animals received an intraperito-
neal injection of carrageenan (20 mg). Three hours
later, the animals were decapitated and the peritoneal
exudate was collected by lavage with PBS [22]. After
centrifugation, the supernatant was used for determina-
tion of NO level, by the method described above [20].
The in vitro release of NO and H2O2 from the
exudate macrophages was also determined. For this,
1 � 106 macrophages from the precipitate were
Fig. 1. Determination of phagocytosis (in vitro zymozan stimulated),
H2O2 and NO (basal, stimulated in vitro by PMA or in vivo by 20 mg
of carrageenan peritoneal) in macrophages of rats from control ( ),
soybean ( ), and fish ( ) groups. Each column represents the
mean T s.e.m. from 9 to 17 animals. (a) Different ( p < 0.05) from
control group, (b) from fish group, and (c) from respective basal.
Fig. 2. NO concentration (mM/100 ml) in peritoneal exudate of rats from
control ( ), soybean ( ) and fish ( ) groups, 3 h after peritoneal
carrageenan stimulus. Each column represents the mean T s.e.m. from 9 to
10 animals. (a) Different ( p < 0.05) from control group.
83Fish/Soybean Oil-Rich Diets Effect on BK, KK, NO, Leptin, Corticosterone and Macrophages in Rats
incubated for 1 h, at 37-C, in PBS containing 5 mM
glucose, 2 mM glutamine, 2% BSA, and 1.3 mM
calcium (incubation solution). The medium was
harvested and NO was measured as described above
[20]. H2O2 in the medium was determined by the
horseradish peroxidase-induced oxidation of phenol red
[23, 24]. Results were normalized for cells total protein
content [25].
For these assays, cell viability was examined by
Trypan blue exclusion (>96%) and cell identity exam-
ined by differential count, confirming that at least 92%
of the peritoneal exudate cells were macrophages.
In vitro Macrophage Stimulation
This procedure was performed in incubated macro-
phages harvested by peritoneal lavage of rats not
stimulated in vivo. The in vitro stimulation consisted
of adding 10 ng of PMA (phorbol myristate acetate) to
the incubation solution, for the 1-h incubation period
described above. NO and H2O2 release were determined
in the medium. Additionally, basal release of NO and
H2O2 was determined by omitting PMA from the
incubation medium of the macrophages from these
non-stimulated rats.
Macrophage Phagocytosis
Phagocytosis was addressed in non-stimulated
macrophages, by adding 1 � 107 particles of opsonized
zymozan to the incubation solution. One hour later, the
percentage of cells presenting at least three zymozan
particles was determined in a counting chamber [26].
Statistical Analysis
Data are expressed as means T S.E.M. Statistical
analysis was performed by KruskallYWallis ANOVA
followed by Dunn test, for the comparisons among the
three diet groups (control, soybean, and fish). The
MannYWhitney test was used for comparisons inside
the same group. Significance level was set at p < 0.05.
RESULTS
Food intake of the fish and soybean groups was
lower than that of the control-diet group (soybean:
25.6% and fish: 29.7%) throughout the seven weeks of
the study. The energy intake was also lower although
not significantly and body weight did not significantly
differ among the groups (data not shown).
Macrophage functionality, as evaluated by both the
phagocytosis of opsonised zymozan particles and the
basal H2O2 production, was not significantly affected by
the consumption of either the soybean or the fish diet. On
the other hand, H2O2 produced by macrophages stimu-
Fig. 3. Paw volume, 3 h after subplantar carrageenan stimulus in
control ( ), soybean ( ) and fish ( ) groups, submitted to
coaxial perfusion. Each column represents the mean T s.e.m. from 8 to
9 animals. (a) Different ( p < 0.05) from control group.
Table 2. Concentration of NO (mM/100 ml) and BK (nmol/ml) on the Paw Edema Perfusate, Collected from First to Second and from Second to
Third Hour, after Subplantar Carrageenan Stimulus, in Rats from Control, Soybean and Fish Groups
NO BK
First to second hour Second to third hour First to second hour Second to third hour
Control 2.25 T 0.68 0.94 T 0.34 58.80 T 3.76 65.93 T 6.77
Soybean 0.77 T 0.35a 0.21 T 0.11 24.14 T 2.87a 21.07 T 7.56a
Fish 0.84 T 0.22 0.09 T 0.05ab 30.68 T 3.38a 29.09 T 6.60a
Data are shown as mean T s.e.m. The number of rats in each group varied between 6 and 11. a Different (p < 0.05) from respective control group andb from respective first to second hour period.
84 Wohlers, Xavier, Oyama, Ribeiro, Nascimento, Casarini, and Silveira
lated either in vitro with PMA or in vivo by the
peritoneal injection of carrageenan, was significantly
decreased in both the lipid groups ( p = 0.0005 and
0.0002, respectively). In fact, after either in vitro or invivo stimulation, only macrophages from the control
group showed a significant increase in the H2O2 produc-
tion, in relation to the respective basal level ( p = 0.02
and p = 0.0005, respectively). The production of NO was
not affected by diet (basal) or in vitro stimulation with
PMA. The in vivo carrageenan stimulus induced incre-
ment in the NO production by peritoneal macrophages
from the control and fish groups ( p = 0.01 for both
groups) (Fig. 1).
Figure 2 shows the levels of NO in the peritoneal
exudate after in vivo stimulation with carrageenan. It
can be seen that NO concentration was lower in both
the soybean and the fish group than in the control group
( p = 0.003).
The paw edema measured 3 h after the subplantar
injection of carrageenan was lower in both soybean and
fish groups than in the control one ( p = 0.0003) (Fig. 3).
NO levels in the paw edema perfusate was also
decreased in the lipid groups in comparison to the
control group. In the soybean group, the decrease was
significant at the first to second-hour period ( p = 0.04)
and in the fish group at the second to third-hour period
( p = 0.007) after carrageenan administration. BK levels
were lower in the perfusate of both hyperlipidic groups
then that of the control rats, in both 1-h collection
periods ( p = 0.0001 and p = 0.001 at the first and
second period, respectively) (Table 2).
The amidolitic activity of plasma KK was also
diminished in the hyperlipidic groups, 1 h ( p = 0.00003)
and 2 h ( p = 0.001) after the carrageenan stimulus
(Fig. 4).
Plasma corticosterone levels did not differ among
the groups during edema development. However, the
basal levels were higher in the lipid groups (Table 3).
The lipid groups showed hyperleptinemia 1 h ( p =
0.003) and 3 h ( p = 0.0004) after the carrageenan
stimulus when compared to the control group. At the
third hour, the leptin levels were higher than at the first
Table 3. Corticosterone Plasma Concentration (mg/100 ml) from 1 to 4 h after Subplantar Carrageenan Stimulus in Rats from Control, Soybean and
Fish Groups
Basal* 1 h* 2 h 3 h 4 h*
Control 11.93 T 0.67 30.72 T 0.75b 31.12 T 1.66b 27.44 T 1.47b 29.16 T 1.46b
Soybean 21.32 T 0.72a 31.35 T 1.06b 29.68 T 1.64b 26.93 T 1.87b 33.74 T 1.49b
Fish 19.45 T 0.53a 32.98 T 0.75b 33.47 T 0.60b 30.88 T 0.89b 29.56 T 2.07b
Data are shown as mean T s.e.m. a Significantly different ( p < 0.05) from respective control group and b from respective basal group. The number of rats
in each group varied between 9 and 15. * Ours previous data from [27], to evaluate the corticosterone release during the carrageenan edema development.
Fig. 4. Amidolitic activity of plasma kalikrein (KK), 1 and 2 hours after
subplantar carrageenan stimulus in control ( ), soybean ( ) and fish
( ) groups. Each column represents the mean T s.e.m from 10 to 12
animals. (a) Different ( p < 0.05) from control group.
85Fish/Soybean Oil-Rich Diets Effect on BK, KK, NO, Leptin, Corticosterone and Macrophages in Rats
hour, in the three groups (control: p = 0.0002; soybean:
p = 0.00002; fish: p = 0.0001) (Table 4).
DISCUSSION
In the present work, we reproduced previous data
from ours and other laboratories, showing that the diets
enriched with either soybean or fish oil decreased food
intake [27Y29]. The reported increased stimulation of
cholecystokinin (CCK) secretion by high-lipid diets may
have contributed to this effect, since CCK is a hormone
involved in the satiety process [30, 31]. The energy
intake, although did not statistically differ among the
groups, tended towards reduction in both the soybean
(Y14%) and the fish group (Y17%) throughout the seven
weeks of the study. Body weight gain was similar among
the groups. This finding has been reported previously
indicating a higher food efficiency for the lipid groups
due the lower energetic cost of lipid deposition from
dietary fat than from dietary carbohydrate [28].
No significant diet effect was evident on macro-
phage phagocytosis, as evaluated 1 h after the addition
of zymozan to the isolated macrophages. However, a
trend towards reduction was seen in both the soybean
(Y17%) and the fish group (Y35%). A decrease in
macrophage phagocytosis has been reported after the
ingestion of nY6 PUFA-rich diet in comparison to the
ingestion of saturated fatty acids-rich diet [32]. In other
reports, however, the consumption of fish oil failed to
change macrophage phagocytic function [15].
The H2O2 production, stimulated by either carra-
geenan or PMA, was decreased in the macrophages
from both hyperlipidic groups, when compared to the
control group. These results are similar to other [33] in
neutrophils from rats fed a soybean-rich diet. H2O2 is an
important citotoxic agent [34] and its lower release
could contribute to the anti-inflammatory effect of the
hyperlipidic diets, as observed previously [12, 27] and
reproduced in the present study.
Some reports have shown decreased activity of
superoxide-dismutase (SOD), the enzyme catalyzing
H2O2 formation, in plasma and in different tissues of
animals treated with nY3 or nY6 PUFA-rich diets [35, 36].
It is possible that a lower SOD activity could be
associated to the low H2O2 production observed in our
study. However, there are controversial information about
the influence of lipid diets on H2O2 production and
antioxidant enzymes activity. Some studies failed to
observe changes on SOD activity in cells obtained from
animals fed with nY3 or nY6 PUFA-rich diets [33, 37]
while others found increase in H2O2 production by
macrophages from rats treated with nY3 PUFA-rich diet
[38]. These different observations could be attributed to
different cell types and quantities of different fatty acids
on the high-lipid diets used. Additionally, the reduction of
H2O2 production, observed in the present study, could
also be attributed to the high basal levels of corticoste-
rone, found in the lipid groups [12], since this hormone
can reduce H2O2 production and activity of antioxidant
enzymes, such as SOD, in macrophages in culture [39].
NO release was decreased in both the peritoneal
exudate and edema perfusate of hyperlipidic groups
stimulated in vivo with carrageenan. NO plays a role in
the carrageenan edema formation [40], promoting vaso-
dilatation and vasopermeability [41]. Thus, its lower
release in the lipid groups likely contributed to the anti-
inflammatory effect of these diets. The observed low BK
levels in the edema perfusate may have influenced NO
release, since it has been demonstrated that BK promotes
NO synthetase (NOs) activation [40]. Additionally, some
studies have shown that both nY3 PUFA [42] and nY6
PUFA-rich diets reduced NOs enzyme [43]. Thus, a low
NOs activity may also have been present in our animals.
This is a possibility to be investigated, since the lipid
groups showed, besides reduced BK release, increased
levels of corticosterone [27], which has been shown to
inhibit NOs [44].
The in vivo results were not equally reproduced in
vitro procedure. The NO production by peritoneal
macrophages from fish group, stimulated in vivo with
carrageenan, was not decreased. This disagreement
could be occurred due the absence Bin vitro^ from
plasma factors, whose interaction must be important for
the in vivo responses. After the PMA stimulus, no
differences were observed in NO production by macro-
phages from all groups, in relation to their respective
baseline. It is possible that the concentration of PMA
and/or the 1-h incubation period were not sufficient to
NO stimulation.
Table 4. Serum Leptin (mg/dl), 1 and 3 h after Subplantar Carra-
geenan Stimulus in Rats from Control, Soybean and Fish Groups
1 h 3 h
Control 1.97 T 0.23 7.82 T 1.34 b
Soybean 5.13 T 0.69 a 15.43 T 1.36 a b
Fish 7.23 T 1.19 a 17.54 T 1.14 a b
Data are shown as mean T s.e.m. a Significantly different (p < 0.05)
from respective control group and b from respective 1 h group. The
number of rats in each group varied between 9 and 14.
86 Wohlers, Xavier, Oyama, Ribeiro, Nascimento, Casarini, and Silveira
BK is an important mediator of the carrageenan
edema [13] that induces hyperalgesia, vasodilatation,
and vasopermeability. We found BK levels to be
diminished in the edema perfusate from the lipid
groups, what is probably related to the reduced kalikrein
levels we found in these animals. At the best of our
knowledge, this is the first demonstration of a causal
association between BK release and the anti-inflamma-
tory effect of fish or soybean oil-rich diets. Since BK is
a product of cininogen cleavage by KK and corticoste-
rone has been shown to decrease cininogen production
[45] the lower BK release could be related to a
reduction on cininogen levels.
It is important to note that other anti-inflammatory
mediators, such as docosatrienes, protectins, and resol-
vins may also be involved, as their production was
reportedly induced by DHA and AA [5]. Additionally,
in view of the reported stimulation of PGE2 release by
BK [46, 47], it is possible that the anti-inflammation
observed in the lipid diets may have relied, at least in
part, on a reduced production of this pro-inflammatory
prostaglandin [48]. Such an effect would be particularly
relevant in the nY6 group, since this diet has been
demonstrated to induce high PGE2 levels [49].
Leptin, a hormone with a role in appetite and energy
expenditure regulation [50], has also been involved in
the inflammatory process, being its production in-
creased, as part of the acute phase response [16, 51, 52].
Leptin levels were higher in the lipid groups, when
compared to control group, probably due to the higher
adiposity induced by the long-term consumption of these
hyperlipidic diets [53]. Additionally, higher corticosterone
basal levels in the lipid groups may have contributed to
the increased leptin, since leptin synthesis has been shown
to be dependent on the presence of glucocorticoids [16].
Increased leptin gene expression has been shown to
follow the intake of hyperlipidic diets, regardless the diet
was rich in nY6 PUFA or saturated fatty acids [54].
Contrarily, a decrease in leptin plasma levels and gene
expression has been reported in rats fed nY3 PUFA, when
compared to rats fed a saturated fatty acids-rich diet [55].
Leptin has been shown to exert a protective effect in
models of gastric injury [56] and to inhibit neutrophils
infiltration and migration in experimental colitis [52, 57].
In contrast, it was suggested a pro-inflammatory role of
leptin in the colitis model [58]. Additionally, leptin-
deficiency has been associated with low in vivo expression
of pro-inflammatory citokines in mice, and leptin addition
to their cultured macrophages increased the expression of
these citokines [59].
In the present study, the higher levels of leptin in the
lipid groups, 1 and 3 h after carrageenan, could be taken as
indicating an anti-inflammatory role of the hormone, since
these groups showed a lower edema in response to
carrageenan. However, the percentage increase of plasma
leptin in the same interval was of 297% in the control
group, while it was of 201 and 143% in the nY6 and nY3
groups, respectively. It is thus possible that the lower
increments, rather than the higher absolute levels, were
relevant, and this assumption would lead to the conclu-
sion of leptin as having a pro-inflammatory action.
Diet enrichment with nY6 PUFA has been associ-
ated with increased levels of inflammatory markers
while dietary supplementation with nY3 PUFA has been
associated with anti-inflammatory effects [60]. The
production of different eicosanoids derived from nY3 or
nY6 PUFA is one of the mechanisms hypothesized to
explain these differences. Indeed, a recent workshop on
the essentiality of these fatty acids advised an increase
of nY3 and a reduction of nY6 PUFA in the diet, to
minimize the adverse effects of AA and its metabolites
[60]. However, there is no clear evidence of the nY6
PUFA prejudicial effects [61]. In contrast, many studies
have been demonstrated beneficial effects of these fatty
acids [62, 63]. It has recently been shown that the
excess of nY6 PUFA failed to antagonize nY3 PUFA
effects and that the association of these two types of
fatty acids lowered the concentration of inflammatory
cytokines, in chronic inflammatory disease [64].
Our previous [12] and present results showed that
nY6 and nY3 PUFA-rich diets had a similar anti-
inflammatory effect and we were presently able to
correlate this effect with similar changes of inflammatory
mediators. The doubt remains as on the real beneficial
effects of reducing the acute inflammation, since the
inflammatory process is a physiological response to
tissue injury, aimed at reestablishing homeostasis.
ACKNOWLEDGMENTS
The authors would like to thank to FAPESP, CAPES
and CNPq for the financial support.
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89Fish/Soybean Oil-Rich Diets Effect on BK, KK, NO, Leptin, Corticosterone and Macrophages in Rats
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