Food Sci. Technol, Campinas, Ahead of Print, 2019 1/7 1
Food Science and Technology
ISSN 0101-2061 (Print)
ISSN 1678-457X (Online)
Ilex paraguariensis A. St. Hil. belongs to the Aquifoliaceae
family and is a native species from the subtropical and temperate
regions of South America. is herb is used regularly in
beverages prepared by infusion such as teas, and is a natural
product, recognized for having anti-inammatory and diuretic
properties (Przygoddaetal., 2010). Likewise, the extract from
Ilex paraguariensis contain dierent bioactive constituents,
and several in vitro studies have shown the important role of
polyphenols on the antioxidant activity, suggesting the potential
use for the development of natural products aiming to protect
biological systems against oxidative stress-mediated damages.
Furthermore, others have shown anti-diabetic and antiobesity
eects of the extract (Kangetal., 2012).
Obesity is an increasing problem worldwide, resulting
in signicant morbidity and mortality, as well as a reduced
quality of life (Hurtetal., 2010). e unbalance on ingestion of
food and loss of energy by exercise causes obesity and visceral
adiposity, promoting complications to the personal health such
as atherosclerosis, hepatic steatosis, and type 2 diabetes (Berg
& Scherer, 2005). Furthermore, the ingestion of a high-fat meal
leads to shis in particle size, numbers, and plasma levels of
very low density lipoprotein (VLDL), low density lipoprotein
(LDL), and high density lipoprotein (HDL) (ALNohair, 2014).
e potential of a diet or food to increase serum concentrations
of cholesterol, especially LDL cholesterol, and promote
atherosclerosis is directly related to its cholesterol, saturated fat
and trans fat content (Menteetal., 2009). Also, it was reported
that consumption of high-fat diets is associated with a reduction
of serum paraoxonase 1 (Pon1) activity (Garciaetal., 2016).
e enzyme Pon1 is a natural antioxidant, and has a primary
role in protecting HDL and LDL from lipid peroxidation (Ngetal.,
2008). Pon1 is synthesized in the liver and secreted into the
bloodstream bound to HDL (Sheetal., 2012). In addition to Pon1,
apolipoprotein A1 (Apoa1) has a specic role in fat metabolism
(Jaichanderetal., 2008), and is a structural protein in HDL, being
the main protein component of HDL (Rader, 2002). Apoa1 is
responsible for the activation of lecithin cholesterol acyltransferase
(LCAT), stimulating cholesterol ow and the binding of HDL to
its receptors (Kontushetal., 2013). In addition to Pon1 and Apoa1,
other genes are involved in lipid metabolism. Fatty acid synthase
(Fas) is the key enzyme required for de novo synthesis of fatty
acids (Wajant, 2012). Stearoyl coenzyme A desaturase-1 (Scd1) is
an enzyme that appears to represent a pivotal control point in lipid
homeostasis. Scd1 catalyzes a rate-limiting step in the biosynthesis of
monounsaturated fats, which are required for triacylglycerol synthesis
and very low density lipoprotein production. In absence of Scd1,
hepatic lipid storage and very low-density lipoprotein production are
impaired, and as default, fatty acids are oxidized (Wajant, 2012). e
peroxisome proliferator-activated receptor co-activator-1 (Pgc1) is
a transcriptional protein co-activator (Handschin, 2010). Pgc1 was
Ilex paraguariensis extract prevents body weight gain in rats fed a high-fat diet
Julia Neitzel UECKER1, Janaina Pereira SCHNEIDER2, Jenifer Heller CERQUEIRA2,
Joao Alveiro Alvarado RINCÓN3, Felipe Teres CAMPOS3, Augusto SCHNEIDER2, Carlos Castilho BARROS2,
Robson ANDREAZZA4, Itiane Barcellos JASKULSKI1, Simone PIENIZ2*
Received 02 Dec., 2017
Accepted 03 Jan., 2019
1 Departamento de Ciência e Tecnologia de Alimentos, Universidade Federal de Pelotas – UFPel, Pelotas, RS, Brasil
2 Departamento de Nutrição, Universidade Federal de Pelotas – UFPel, Pelotas, RS, Brasil
3 Departamento de Clínicas Veterinárias, Universidade Federal de Pelotas – UFPel, Pelotas, RS, Brasil
4 Centro de Engenharias, Universidade Federal de Pelotas – UFPel, Pelotas, RS, Brasil
*Corresponding author: firstname.lastname@example.org
Studies have shown that drinks containing Ilex paraguariensis extract can promote many benets in animals and in humans.
e present study aimed to evaluate in vivo eects of Ilex paraguariensis extract on metabolic prole, obesity prevention and
expression of genes related with adipogenesis and lipogenesis in Wistar female rats fed a high-fat diet. For this experiment
32 Wistar female rats with normal weight were used and randomly separated into four groups: diet (standard or high-fat)
and treatment (water or Ilex paraguariensis extract) for 34 days. e rats receiving Ilex paraguariensis extract had lower body
weight compared to the control group in both diets. Likewise, there was a reduction in triglycerides in the groups fed high-fat
diet and treated with Ilex paraguariensis extract. e creatinine levels were lower in the groups treated with Ilex paraguariensis
and in high-fat diet. It was observed an increased liver gene expression for Fas and Scd1 in the group treated with hyperlipid
diet + Ilex paraguariensis. It can be concluded that Ilex paraguariensis extract decreased body weight gain in both control and
high-fat diets, reduced plasma triglycerides and creatinine levels and increased liver expression of genes related to lipogenesis.
Keywords: obesity; triglycerides; glucose levels; genes.
Practical Application: Control of obesity by the consumption of the extract of the Ilex paraguariensis.
Food Sci. Technol, Campinas, Ahead of Print, 20192 2/7
Ilex paraguariensis prevents obesity
initially characterized in adipose tissue and it is now known that
this molecule plays an important role in oxidative metabolism,
in mitochondrial biogenesis and hepatic gluconeogenesis. Based
on this, evaluation o liver expression of these genes represents an
important hallmark of the eect of the high-fat diets.
us, to evaluate the ecacy of potential compounds in the
prevention and treatment of obesity, several animal models have
been used (Kangetal., 2012). It has been reported that rodents
fed with a high-fat diet are a good model of obesity, where the
dietary environment is a major contributor (Bulloetal., 2007).
Based on these evidences, this study aimed to evaluate the eects
of Ilex paraguariensis extract on metabolic prole, body weight
gain and liver gene expression related with adipogenesis and
lipogenesis of Wistar female rats fed to a high-fat diet.
2 Materials and methods
2.1 Experimental conditions and monitoring
e experimental protocol was approved by the Animal
Welfare Commission from the Federal University of Pelotas
(Rio Grande do Sul State, Brazil), under the number 1641, and
all procedures were conducted according to the guidelines of
laboratory animal use in research. For this study 32 female Wistar
rats (Rattus Novergicus), 60 days old were used. e rats were kept
in groups of four at polypropylene boxes in ventilated cabinets,
with controlled temperature and relative humidity conditions
(23 °C ± 1 °C and 65-75%), and exposed to a 12 hour light/dark
cycle. Aer ve days of adaptation, the rats were randomly divided
into four groups (n= 8 rats per group), as follows: standard diet
(4% fat) + water ad libitum (SW); standard diet (4% fat) + Ilex
paraguariensis extract ad libitum (SIP); high-fat diet (25% fat,
1% cholesterol and 0.1% cholic acid) + water ad libitum (HFW);
high-fat diet (25% fat, 1% cholesterol and 0.1% cholic acid) + Ilex
paraguariensis extract ad libitum (HFIP). e standard (4% fat
content) and high-fat diets (25% fat content, 1% cholesterol and
0.1% cholic acid) were prepared in the laboratory, as recommended
by the American Institute of Nutrition - AIN93- M for rodents
(Reevesetal., 1993) (Table1). e same brand and batch of Ilex
paraguariensis was used throughout the experiment period,
guaranteeing product homogeneity. e Ilex paraguariensis extract
was prepared in a 10% concentration and 70 °C temperature,
resembling conditions of the human ingestion. e infusion of
the extract was performed for 20 minutes and aer it was sieved.
e body weight was measured weekly using an electronic
scale (JH2102/Bioprecisa, Curitiba, Brazil) and food intake
was monitored daily during the 34 days of the study. Rats
were euthanized at day 34, aer a 12 hours fasting, following
the ethical principles in animal experimentation used by the
Brazilian College of Animal Experimentation.
2.2 Sample collection and biochemistry analysis
Blood was collected and centrifuged at 1000rpm for 10 minutes
(Centrifuge 5415, Eppendorf, Westbury, New York, USA).
e serum was transferred to a microtube and frozen at -20 °C
e determination of total cholesterol, HDL, triglycerides
and glucose levels in serum were performed using commercial
kits (Triglycerides Liquiform and Glucose Liquiform, respectively,
, Minas Gerais, Brazil). Samples reading were performed
in a spectrophotometer (Ultraspec 2000, Pharmacia Biotech)
at 500 nm. e results from total cholesterol, HDL cholesterol,
triglycerides and glucose levels were expressed in mg/dL. e intra
assay coecients of variation found between 11.4% and 17.0%.
e Pon1 activity was measured by its arylesterase activity as
previously established (Sheetal., 2012). e arylesterase activity
was measured by the phenol formation rate through monitoring
the increase in absorbance at 270 nm and 25 °C. e working
reagent consisted of 20 mM Tris/HCl, pH 8.0, containing 1mM
and 4 mM phenylacetate as substrate. e samples
were diluted 1:3 in a 20 mM Tris/HCl buer and were added
to the working reagent and the change in absorbance recorded
for 60 sec. e activity the Pon1 was expressed in U/L, based on
the phenol extinction coecient. e intra assay coecients of
variation found were 14.7%.
e determination of the concentration of transaminases
glutamic-oxaloacetic transaminase (GOT) and glutamic-pyruvic
transaminase (GPT) in the serum was performed by colorimetric
method using a commercial kit (Doles, Goiânia - GO, Brazil)
and readings obtained spectrophotometrically at 505 nm and
the results expressed as IU/L.
e determination of creatinine concentrations in serum
were performed using commercial kits (Creatinine K, Labtest
Diagnostica SA, Lagoa Santa, Brazil) based on the Jaé reaction.
For the measurements, 50 uL of serum sample was mixed with
50 uL of alkaline picrate. Subsequently, the reading was held in
spectrophotometer at 520 nm aer 0 and 60 seconds. e results
were expressed in mg/dL.
2.3 Gene expression
To determine gene expression, samples of liver were collected
and immediately frozen in liquid nitrogen and stored at -80oC.
e samples were homogenized with Qiazol (Qiagen, Valencia,
USA), and total RNA was isolated and puried following the Qiazol
protocol. e quality of RNA was assessed by electrophoresis in
agarose gel. e reverse transcription reactions were performed
using 1 µg of RNA with a reverse transcription kit containing
RNase inhibitor (Applied Biosystems, Foster City, USA) in a
volume of 10 µL. Real-time PCR was performed to assess the
Ta bl e 1 . Diet composition according to AIN93-M (Reevesetal., 1993).
Standard Diet High-fat Diet
g.kg -1 g.kg -1
Maize starch 466 256
Casein 140 140
Saccharose 100 100
Dextrinated starch 155 155
Soy oil 40 40
Fiber 50 50
Minerals 35 35
Vitamins 10 10
L-cistina 1.8 1.8
β-Colina 2.5 2.5
Tetra Butilh 0.008 0.008
Lard ---- 120
Overall 1000 1000
Food Sci. Technol, Campinas, Ahead of Print, 2019 3/7 3
expression of the target genes Apoa1, Pon1, Scd1, Foxo, Fasn
and Pgc1 and the internal control Actb (Table2).
PCR reactions were performed in duplicate in a volume of
12 µL using SYBR Green Mastermix (Applied Biosystems) and the
uorescence was quantied in the Eco Real Time (Illumina, San
Diego, California, USA). For each test, 40 cycles were carried out
and a dissociation curve was included at the end of the reaction in
order to verify the amplication of a single PCR product. Data is
reported as folds over the minimum according to Masternaketal.
(2005). Each assay plate included a negative control with water.
2.4 Statistical analysis
Data was analyzed using two-way analysis of variance (Two-way
ANOVA) and Tukey’s test at 5% signicance level for comparison of
means, using Graphpad Prism 5.0 (GraphPad, La Jolla, CA, USA).
e eects of the diet, supplementation with Ilex paraguaiensis
extract and their interaction were tested. When the interaction
was signicant individual groups were compared by t-test.
e results from feed intake indicate that the groups fed
high-fat diet had lower intake (p < 0.01, Table3) when compared
to the standard diet groups, although there was no eect of
Ilex paraguariensis extract supplementation nor interaction
between the diet and treatment (p > 0.05, Table3). Despite no
dierence in feed intake when submitted to the same diet, rats
supplemented with Ilex paraguariensis extract presented smaller
weight gain (p < 0.01). No eect of diet or interaction between Ilex
paraguariensis extract and diet was observed (p > 0.05, Table3).
Ilex paraguariensis extract supplementation was eective to
reduce plasma triglycerides on groups submitted to both diets
(standard or high-fat diet) (p < 0.05, Table4). e analysis from
Tab le 2. Primers used in the analysis of gene expression by real-time PCR.
Primers Gene Sequence (5’-3’) Length (pb)
Actb (forward) β-actin TCACCACCACAGCCGAGAGA 72
Actb (reverse) CGAAATCCAGTGCGACGTAGC
Pon1 (forward) Pon1 CAAGAACCATCGGTCTTCCT 197
Actb (reverse) CAGGCTTACTGGGATCGAAA
Apoa1 (forward) Apoa1 CAAGAACCATCGGTCTTCCT 828
Apoa1 (reverse) CAGGCTTACTGGGATCGAAA
Scd1 (forward) Scd1 CACATCAACTTCACCACGTTCTTC 74
Scd1 (reverse) GAAACTTTCTTCCGGTCGTAAGC
Foxo3 (forward) Foxo3 TCCCAGATCTACGAGTGGATGG 42
Foxo3 (reverse) CCTTCATTCTGAACGCGCAT
Fasn (forward) FAS CTGGACTCGCTCATGGGTG 111
Fasn (reverse) CATTTCCTGAAGTTTCCGCAG
Pgc1 (forward) PGC1 CCGAGAATTCATGGAGCAAT 114
Pgc1 (reverse) TTTCTGTGGGTTTGGTGTGA
Tab l e 3 . Analysis of food intake and average weight of female rats in control (SW and HFW) and Ilex paraguaiensis extract groups (SIP and HFIP).
e values were expressed as mean (M) ± standard error (SE).
Groups Feed intake (g) Total caloric intake Starting weight (g) Final weight (g) Weight gain (g)
----- M ± SE ----- ----- Kcal ----- ----- M ± SE -----
SW* 22.31 ± 0.39a** 3624.8 194.89 ± 0.43 233.38 ± 0.21 38.49 ± 0.23a**
SIP 21.85 ± 0.27a3624.8 191.25 ± 0.36 222.58 ± 0.26 31.33 ± 0.31 b
HFW 19.36 ± 0.34b3907.1 192.28 ± 0.54 238.94 ± 0.78 46.66 ± 3.03a
HFIP 19.85 ± 0.24b3907.1 192.37 ± 0.41 225.41 ± 0.62 33.04 ± 0.45 b
CV(%) 11.12 ----- 12.35 11.97 11.20
*SW: standard diet (4% fat) + water ad libitum; SIP: standard diet (4% fat) + Ilex paraguariensis extract ad libitum; HFW: high-fat diet (25% fat content, 1% cholesterol and 0.1% cholic
acid) + water ad libitum; HFIP: high-fat diet (25% fat content, 1% cholesterol and 0.1% cholic acid) + Ilex paraguariensis extract ad libitum; CV: coecient variation; **Average values
followed by the same letter in the column show no signicant statistical dierence between them with the Tukey test at 1% probability of error (p < 0.01).
Tab l e 4 . Analysis of total cholesterol, HDL cholesterol, triglycerides levels, blood glucose and Pon1 activity of female rats in the control and
treatment groups. e values were expressed as mean (M) ± standard error (SE).
Treatment Total cholesterol HDL cholesterol Triglyceride Glucose Pon1
----- mg/dL ----- ----- U/L -----
SW* 73.45 ± 04.90 ns** 53.58 ± 07.05 ns 117.76 ± 05.72 a*** 464.76 ± 37.19 ns 134.10 ± 10.73 ns
SIP 81.74 ± 08.73 ns 60.68 ± 05.67 ns 92.55 ± 02.31 ab 399.29 ± 25.69 ns 151.95 ±10.62 ns
HFW 80.99 ± 07.23 ns 60.19 ± 06.90 ns 89.50 ± 03.18 ab 370.72 ± 16.48 ns 125.30 ±13,81 ns
HFIP 78.80 ± 11.32 ns 51.39 ± 04.01 ns 64.10 ± 01.80 b390.72 ± 35.12 ns 132.10 ±10.19 ns
CV(%) 11.35 16.78 11.55 16.99 14.71
*SW: control diet (4% fat) + water ad libitum; SIP: control diet (4% fat) + Ilex paraguariensis extract ad libitum; HFW: fat diet (25% fat content, 1% cholesterol and 0.1% cholic acid) + water
ad libtum; HFIP: fat diet (25% fat content, 1% cholesterol and 0.1% cholic acid) + Ilex paraguariensis extract ad libitum; CV: coecient variation; **not signicant; ***Average values
followed by the same letter show no signicant statistical dierence between them with the Tukey test at 1% probability of error (p < 0.01).
Food Sci. Technol, Campinas, Ahead of Print, 20194 4/7
Ilex paraguariensis prevents obesity
total cholesterol, HDL cholesterol and glucose concentration
indicates no signicant changes induced by diet, Ilex paraguariensis
or its interaction (p > 0.01, Table4). Likewise, Pon1 activity
was not aected by diet, Ilex paraguariensis or its interaction
(p > 0.05, Table4).
ere was no dierence in transaminase enzymes GOT
(Figure1A) and GPT (Figure1B) activity between groups
(p > 0.05), indicating no change in liver function for groups
treated with Ilex paraguariensis extract.
Creatinine levels (Figure1C) were lower in the group
treated with standard diet + Ilex paraguariensis extract when
compared to the standard diet + water group (p < 0.01). When
only the rats fed the high-fat diet were analyzed, no dierence
(p>0.05) was observed between the groups treated with water
or Ilex paraguariensis extract.
Regarding liver gene expression, Apoa1 (Figure2A), Pon1
(Figure2B), Foxo3 (Figure2E) and Pgc1 (Figure2F) expression
were not dierent between groups. However, Fasn (Figure2C)
and Scd1 (Figure2D) were higher (p < 0.01) in the high-fat + Ilex
We observed lower food intake in the high-fat compared
to the standard groups. Kojima & Kangawa, (2005) reported
that consumption of the high-fat diet induced a satiating eect,
which is reected in the reduced levels of the appetite-stimulating
peptide ghrelin. Thomàs-Moyàetal. (2007) reported that
rats feed with high-fat diet reduced their food intake, thus
maintaining their energy intake and their body weight closer
to those of the control rats. However, a marked increment of
adipose depots was observed, which was greater in males than
females (Thomàs-Moyàetal., 2007). e reduced intake in rats
fed a high-fat diet is well known, and Haririetal. (2010) found
Figure 1. Analysis of the GOT (A) and GPT (B) enzymes, and creatinine
(C) determined by colorimetric method. Results were expressed in
mean ± standard error. SW: standard diet + water; SIP: standard
diet + Ilex paraguariensis extract; HFW: hyperlipidic diet + water;
HFIP: hyperlipidic diet + Ilex paraguariensis extract.
Figure 2. Ilex paraguariensis extract intake eect of gene expression of Apoa1 (A), Pon1 (B), Fas (C), Sdc1 (D), Foxo3a (E), Pgc1 (F), in the
liver of female rats supplemented with standard and high-fat diet. SW: standard diet + water; SIP: standard diet + Ilex paraguariensis extract;
HFW: hyperlipidic diet + water; HFIP: hyperlipidic diet + Ilex paraguariensis extract.
Food Sci. Technol, Campinas, Ahead of Print, 2019 5/7 5
similar results regarding food intake, when feeding a high-fat
diet to rats for a period of 26 days, in order to analyze the eect
of diet as a facilitator to excessive weight gain.
Body weight gain was lower in rats supplemented with
Ilex paraguariensis extract. Similar data was observed by
Panget al. (2008), indicating that dietary supplementation
with Ilex paraguariensis extract administered to obese rats
induced by high-fat diet was able to signicantly reduce body
weight gain, plasma triacylglycerides, glucose,, along with
reduction on anti-inammatory markers (Luzet al., 2016),
and antidepressant-like eects (Reisetal., 2014). According
to Hetzleretal. (1990) the amount of caeine present in Ilex
paraguariensis could be responsible for the signicant decrease in
the amount of epididymal and abdominal fat. e caeine has been
shown to be able to cross the blood brain barrier and to increase
the circulating concentrations of catecholamine (epinephrine)
in humans, which is known to increase thermogenesis and
lipolysis (Silvaetal., 2011). In studies with mice fed a high-fat
diet, Ilex paraguariensis has been suggested to promote satiety
through several mechanisms, including induction and/or
enhancement of intestinal glucagon-like peptide-1 (GLP-1),
modulation of serum leptin levels and a possible direct central
satiety-stimulatory eect (Resendeetal., 2012). Data obtained
from experiments conducted in diet-induced obesity models
have shown that Ilex paraguariensis suppresses body weight
gain and visceral fat accumulation and decreases serum levels
of cholesterol, triglycerides, LDL cholesterol, glucose, insulin,
pancreatic lipase and leptin (Gambero & Ribeiro, 2015). erefore,
our study further support the idea that Ilex paraguariensis
extract treatment is able to reduce body weight gain in rats fed
a high-fat diet.
We also observed that serum triglycerides were lower for
rats fed high-fat diet and receiving Ilex paraguariensis extract
than rats fed high-fat diet and receiving only water. Silvaetal.
(2011) reported, for rats that consumed the extract of gross
mate, an increase in triglyceride levels of 21.4% compared to
the control group. Increased triglyceride levels may be related
to the lipolytic eect, resulting in increased mobilization of fatty
acids from adipose tissue or intramuscular fat depots. However,
Paganini-Steinetal. (2005) found in their study that animals
treated with Ilex paraguariensis had a decrease in triglyceride
levels compared to controls animals, in agreement with our
current ndings. Another study with mice treated with high-fat
diet and Ilex paraguariensis also showed that Ilex paraguariensis
reduced plasma triglycerides (Kangetal., 2012). erefore, in
addition to reducing body weight gain, Ilex paraguariensis results
in decreased triglycerides levels which can be benecial in the
prevention of heart diseases.
e present study indicated that total cholesterol, HDL
and Pon1 activity were not aected by diet, Ilex paraguariensis
extract or its interaction. Recently, Bravoetal. (2014) studying
the eect of Ilex paraguariensis on serum lipids and antioxidant
status of normocholesterolemic and hypercholesterolemic rats
demonstrated that in the normocholesterolemic rats, the Ilex
paraguariensis had no eect on serum lipids or antioxidant status.
Despite that in the hypercholesterolemic rats, Ilex paraguariensis
treatment also had no eect on HDL-c or protein carbonyls, it
showed a marked hypolipidemic action, decreasing triglycerides,
total cholesterol and LDL-c, and serum malonaldehyde levels.
ese parameters had been increased aer consumption of a
high cholesterol diet, pointing out that the potential benec
eect of Ilex paraguariensis on risk factors for cardiovascular
diseases seems to be restricted to already hyperlipidemic
animals. Paganini-Steinetal. (2005) were the rst to report a
signicant reduction in serum total cholesterol and triglycerides
of cholesterol-fed rats aer administration of Ilex paraguariensis
aqueous extract. Thomàs-Moyàetal. (2007) also reported that
Ilex paraguariensis extract decreased the VLDL-LDL fraction in
obese rats. A probable mechanism for the LDL-C lowering ability
of Ilex paraguariensis is the blocking of cholesterol absorption in
the small intestine and/or the inhibition of cholesterol synthesis
in the liver, which can be attributed to the presence of saponins,
phenolic compounds, avonoids, and/or caeine in the mate
infusion (Moraisetal., 2009).
e present study also analyzed the GOT and GPT enzymes,
which were not dierent among groups. ese enzymes have
been investigated in order to verify possible liver damages
induced by the high-fat diet. ey are found within cells in the
liver, but when there is some abnormal liver function, lead to
an increase in these enzymes, which are released into the blood
stream. Usually this increase is asymptomatic and transient, but
some diseases caused by elevations of GOT and GPT levels are
acute hepatitis A or B, fatty liver, obesity and hepatitis C (Russo
& Jacobson, 2012). Despite that, creatinine was lower in the
groups treated with Ilex paraguariensis. Creatinine has been
used as a parameter for the initial evaluation of renal function in
daily clinical practice. However, inferring that normal creatinine
values always indicate normal kidney function can lead to
signicant errors, since early changes in glomerular ltration
rate can be “hidden” in normal’s creatinine values (Pintoetal.,
2004). erefore, higher creatinine level can reveal that renal
function is disturbed, indicating that Ilex paraguariensis can be
benecial for kidney function.
e eects of Ilex paraguariensis extract on the gene
expression of antioxidant/inammatory markers have been
studied in animal models (Matsumotoetal., 2009; Arçarietal.,
2011). e analysis of liver tissue expression of Pon1 and
Apoa1, indicated no dierence between diets and treatments.
Similar results were demonstrated by Boaventuraetal. (2012)
in humans, indicating that there was no change in Pon1 activity
aer a prolonged ingestion (90 days) of Ilex paraguariensis tea.
However, Meninietal. (2007) reported increases in serum Pon1
activity aer ingesting 500 mL of Ilex paraguariensis infusion in
healthy individuals. Bastos & Gugliucci (2009) demonstrated that
the chlorogenic acid, the main phenolic constituent of the Ilex
paraguariensis, can preserve Pon1 against oxidative degradation in
vitro. Results found by Moraisetal. (2009) in humans corroborate
with the data in this study, and also did not observe signicant
increases in Apoa1 expression, suggesting that this can be due
to the decreased HDL catabolism. e Apoa1 is indicative of the
amount of HDL in plasma or new HDL particles forming the
potential to exert its function in reverse cholesterol transport
to the liver. When there is an increase in Apoa1, it is suggested
that there is an increase in hepatic production of nascent HDL
particles (Lyssenkoetal., 2013).
Food Sci. Technol, Campinas, Ahead of Print, 20196 6/7
Ilex paraguariensis prevents obesity
Diet-induced obesity is largely caused by disorders of fat
metabolism, resulting in a massive accumulation of fat in various
tissues. Lipid and energy metabolism are regulated by a complex
network of signaling processes, therefore we investigated the
mRNA expression of key genes regulating lipid metabolism
such as Scd1 and Fasn (Yangetal., 2012). Fasn, which encodes
a rate limiting enzyme in fatty acid biosynthesis to produce
palmitic acid; Scd1, which converts stearic acid to oleic acid, and
glycerol-3-phosphate acyltransferase, which encodes the rst
committed enzyme in triglyceride and phospholipid synthesis
(Hortonet al., 2002). e mRNA expression levels of genes
encoding lipogenic proteins such as Scd1 and Fas increased in
rats receiving Ilex paraguaiensis and high-fat diet, can be suggest
a rapid eect of the high-fat diet on lipogenesis. Expression of
Foxo gene, which regulates gluconeogenesis, and Pgc1 gene,
a coactivator essential for coordinating gluconeogenesis and
fatty acid oxidation, were not dierent among groups in this
study. Additionally, Pon1 and Apoa1 gene expression were also
not dierent, conrming the observations that serum levels of
Pon1 were not changed as well as of HDL.
In summary, the data presented here indicates that the
use of Ilex paraguariensis extract prevents body weight gain,
while improving the lipid parameters in rats fed a high-fat diet.
In addition, Ilex paraguariensis modulates the expression of genes
related in adipogenesis and lipogenesis in the obese state. us,
the results from this study indicated that Ilex paraguariensis
extract might be helpful in the treatment against obesity and
is study was nanced in part by the Coordenação de
Aperfeiçoamento de Pessoal de Nível Superior – Brasil (CAPES)
– Finance Code 001. Authors thank also the nancial support of
the Brazilian agencies: Conselho Nacional de Desenvolvimento
Cientíco e Tecnológico – Brasil (CNPq) and Fundação de
Amparo à Pesquisa do Estado do Rio Grande do Sul – Brasil
ALNohair, S. (2014). Obesity in gulf countries. International Journal
of Health Sciences, 8(1), 79-83. http://dx.doi.org/10.12816/0006074.
Arçari, D. P., Bartchewsky, W. Jr., Santos, T. W., Oliveira, K. A., Oliveira,
C. C., Gotardo, É. M., Pedrazzoli, J. Jr., Gambero, A., Ferraz, F.
C., Carvalho, P. O., & Ribeiro, M. L. (2011). Anti-inflammatory
effects of yerba maté extract (Ilex paraguariensis) ameliorate insulin
resistance in mice with high fat diet-induced obesity. Mole cular and
Cellular Endocrinology, 335(2), 110-115. http://dx.doi.org/10.1016/j.
Bastos, D. H., & Gugliucci, A. (2009). Chlorogenic acid protects
paraoxonase 1 activity in high density lipoprotein from inactivation
caused by physiological concentrations of hypochlorite. Fitoterapia,
80(2), 138-142. http://dx.doi.org/10.1016/j.fitote.2009.01.001.
Berg, A. H., & Scherer, P. E. (2005). Adipose tissue, inflammation, and
cardiovascular disease. Circulation R esearch, 96(9), 939-949. http://
Boaventura, B. C. B., Di Pietro, P. F., Stefanuto, A., Klein, G. A., Morais,
E. C., Andrade, F., Wazlawik, E., & Silva, E. L. (2012). Association
of mate tea (Ilex paraguariensis) intake and dietary intervention
and effects on oxidative stress biomarkers of dyslipidemic subjects.
Nutrition (Burbank, Los Angeles County, Calif.), 28(6), 657-664. http://
Bravo, L., Mateos, R., Sarriá, B., Baeza, G., Lecumberri, E., Ramos, S.,
& Goya, L. (2014). Hypocholesterolaemic and antioxidant effects
of yerba mate (Ilex paraguariensis) in high-cholesterol fed rats.
Fitoterapia, 92, 219-229. http://dx.doi.org/10.1016/j.fitote.2013.11.007.
Bullo, M., Casas-Agustench, P., Amigo-Correig, P., Aranceta, J., &
Salas-Salvado, J. (2007). Inflammation, obesity and comorbidities:
the role of diet. Public Health Nutrition, 10(10A), 1164-1172. http://
Gambero, A., & Ribeiro, M. L. (2015). The positive effects of yerba
mate (Ilex paraguariensis) in obesity. Nutrients, 7(2), 730-750. http://
Garcia, D. N., Prietsch, L. A., Rincón, J. A., Moreira, I. L., Valle, S. C.,
Barros, C. C., Helbig, E., Corrêa, M. N., & Schneider, A. (2016).
Differential effects of a high-fat diet on serum lipid parameters
and ovarian gene expression in young and aged female mice. Zygote
(Cambridge, England), 24(5), 676-683. http://dx.doi.org/10.1017/
Handschin, C. (2010). Regulation of skeletal muscle cell plasticity by
the peroxisome proliferator-activated receptor coactivator 1. Journal
of Receptors and Signal Transduction, 30(6), 376-384. http://dx.doi.
Hariri, N., Gougeon, R., & Thibault, L. (2010). A highly saturated fat-rich
diet is more obesogenic than diets with lower saturated fat content.
Nutrition Research (New York, N.Y.), 30(9), 632-643. http://dx.doi.
Hetzler, R. K., Knowlton, R. T., S omani, S. M., Brown, D. D., & Perkins,
R. M. 3rd (1990). Effect of paraxanthine on FFA mobilization after
intravenous caffeine administration in humans. Journal of Applied
Physiology, 68(1), 44-47. http://dx.doi.org/10.1152/jappl.19184.108.40.206.
Horton, J. D., Goldstein, J. L., & Brown, M. S. (2002). SREBPs: activators
of the complete program of cholesterol and fatty acid synthesis in
the liver. e Journal of Clinical Investigation, 109(9), 1125-1131.
Hurt, R. T., Kulisek, C., Buchanan, L. A., & McClave, S. A. (2010). The
obesity epidemic: challenges, health initiatives, and implications for
gastroenterologists. Journal of Gastroenterology and Hepatology,
6(12), 780-792. PMid:21301632.
Jaichander, P., Selvarajan, K., Garelnabi, M., & Parthasarathy, S. (2008).
Induction of paraoxonase 1 and apolipoprotein A-I gene expression
by aspirin. Journal of Lipid Research, 49(10), 2142-2148. http://dx.doi.
Kang, Y.-R., Lee, H.-Y., Kim, J.-H., Moon, D.-I., Seo, M.-Y., Park, S.-H.,
Choi, K.-H., Kim, C.-R., Kim, S.-H., Oh, J.-H., Cho, S.-W., Kim,
S.-Y., Kim, M.-G., Chae, S.-W., Kim, O., & Oh, H.-G. (2012). Anti-
obesity and anti-diabetic effects of yerba mate (Ilex paraguariensis) in
C57BL/6J mice fed a high-fat diet. Laboratory Animal Research, 28(1),
23-29. http://dx.doi.org/10.5625/lar.2012.28.1.23. PMid:22474471.
Kojima, M., & Kangawa, K. (2005). Ghrelin: structure and function.
Physiological Reviews, 85(2), 495-522. http://dx.doi.org/10.1152/
Food Sci. Technol, Campinas, Ahead of Print, 2019 7/7 7
tissue. Archives of Biochemistry and Biophysics, 476(2), 178-185.
Pinto, P. S., Silva, F. J., & Munch, E. C. S. M. (2004). Inadequabilidade
da creatinina sérica na identificação precoce da disfunção renal.
Jornal Brasileiro de Neurologia, 26, 196-201.
Przygodda, F., Martins, Z. N., Castaldelli, A. P. A., Minella, T. V., Vieira, L.
P., Cantelli, K., Fronza, J., & Padoin, M. J. (2010). Effect of erva-mate
(Ilex paraguariensis A. St.-Hil., Aquifoliaceae) on serum cholesterol,
triacylglycerides and glucose in Wistar rats fed a diet supplemented
with fat and sugar. Brazilian. Journal of Pharmacognsosy, 20(6),
Rader, D. J. (2002). High-density lipoproteins and atherosclerosis.
e American Journal of Cardiology, 90(8A), 62-70. http://dx.doi.
Reeves, P. G., Nielsen, F. H., & Fahey, G. C. Jr. (1993). AIN-93 purified
diets for laboratory rodents: final report of the American Institute
of Nutrition ad hoc writing committee on the reformulation of the
AIN-76A rodent diet. Journal of Nutrition, 123(11), 1939-1951.
Reis, E. M., Schreiner, F. W. No., Cattani, V. B., Peroza, L. R., Busanello,
A., Leal, C. Q., Boligon, A. A., Lehmen, T. F., Libardoni, M., Athayde,
M. L., & Fachinetto, R. (2014). Antidepressant-Like Effect of Ilex
paraguariensis in Rats. BioMed Research International, 1, 1-9. http://
Resende, P. E., Verza, S. G., Kaiser, S., Gomes, L. F., Kucharski, L.
C., & Ortega, G. G. (2012). The activity of mate saponins (Ilex
paraguariensis) in intra-abdominal and epididymal fat, and glucose
oxidation in male Wistar rat. Journal of Ethnopharmacology, 144(3),
735-740. http://dx.doi.org/10.1016/j.jep.2012.10.023. PMid:23088849.
Russo, M. W., & Jacobson, I. M. (2012). How to use statins in patients
with chronic liver disease. Journal of Medicinal Chemistry, 71(1),
She, Z. G., Chen, H. Z., Yan, Y., Li, H., & Liu, D. P. (2012). The human
paraoxonase gene cluster as a target in the treatment of atherosclerosis.
Antioxidants & Redox Signalling, 16(6), 597-632. http://dx.doi.
Silva, R. D., Bueno, A. L. S., Gallon, C. W., Gomes, L. F., Kaiser, S.,
Pavei, C., Ortega, G. G., Kucharski, L. C., & Jahn, M. P. (2011). The
effect of aqueous extract of gross and commercial yerba mate (Ilex
paraguariensis) on intra-abdominal and epididymal fat and glucose
levels in male Wistar rats. Fitoterapia, 82(6), 818-826. http://dx.doi.
Thomàs-Moyà, E., Gianotti, M., Proenza, A. M., & Lladó, I. (2007).
Paraoxonase 1 response to a high-fat diet: gender differences in the
factors involved. Journal of Molecular Medicine (Berlin, Germany),
13(3-4), 203-209. PMid:17592556.
Wajant, H. (2012). The Fas signaling pathway: more than a paradigm.
Science, 296(5573), 1635-1636. http://dx.doi.org/10.1126/
Yang, Z. H., Miyahara, H., Takeo, J., & Katayama, M. (2012). Diet high
in fat and sucrose induces rapid onset of obesity-related metabolic
syndrome partly through rapid response of genes involved in
lipogenesis, insulin signalling and inflammation in mice. Diabetology
& Metabolic Syndrome, 4(1), 1-10. http://dx.doi.org/10.1186/1758-
Kontush, A., Lhomme, M., & Chapman, M. J. (2013). Unraveling the
complexities of the HDL lipidome. Journal of Lipid Research, 54(11),
Luz, A. B. G., Silva, C. H. B., Nascimento, M. V. P. S., Facchin, B. M. C.,
Baratto, B., Fröde, T. S., Reginatto, F. H., & Dalmarco, E. M. (2016).
The anti-inflammatory effect of Ilex paraguariensis A. St. Hil (Mate)
in a murine model of pleurisy. International Immunopharmacology,
36, 165-172. http://dx.doi.org/10.1016/j.intimp.2016.04.027.
Lyssenko, N. N., Nickel, M., Tang, C., & Phillips, M. C. (2013). Factors
controlling nascent high-density lipoprotein particle heterogeneity:
ATP-binding cassette transporter A1 activity and cell lipid and
apolipoprotein AI availability. e FASEB Journal, 27(7), 2880-2892.
Masternak, M. M., Al-Regaiey, K. A., Del-Rosario-Lim, M. M.,
Bonkowski, M. S., Panici, J. A., Przybylski, G. K., & Bartke, A. (2005).
Caloric restriction results in decreased expression of peroxisome
proliferator-activated receptor super family in muscle of normal
and long-lived growth hormone receptor/binding protein knockout
mice. e Journals of Gerontology. Series A, Biological Sciences and
Medical Sciences, 60(10), 1238-1245. http://dx.doi.org/10.1093/
Matsumoto, R. L., Bastos, D. H., Mendonça, S., Nunes, V. S., Bartchewsky,
W., Ribeiro, M. L., & Carvalho, P. O. (2009). Effects of mate tea
(Ilex paraguariensis) ingestion on mRNA expression of antioxidant
enzymes, lipid peroxidation, and total antioxidant status in healthy
young women. Journal of Agricultural and Food Chemistry, 57(5),
1775-1780. http://dx.doi.org/10.1021/jf803096g. PMid:19219987.
Menini, T., Heck, C., Schulze, J., Mejia, E., & Gugliucci, A. (2007).
Protective action of Ilex paraguariensis extract against free radical
inactivation of paraoxonase-1 in high-density lipoprotein. Planta
Medica, 73(11), 1141-1147. http://dx.doi.org/10.1055/s-2007-981585.
Mente, A., Koning, L., Shannon, H. S., & Anand, S. S. (2009). A systematic
review of the evidence supporting a causal link between dietary
factors and coronary heart disease. Archives of Internal Medicine,
169(7), 659-669. http://dx.doi.org/10.1001/archinternmed.2009.38.
Morais, E. C., Stefanuto, A., Klein, G. A., Boaventura, B. C. B., Andrade,
F., Wazlawik, E., Pietro, P. F., Maraschin, M., & Silva, E. L. (2009).
Consumption of yerba mate (Ilex paraguariensis) improves serum
lipid parameters in healthy dyslipidemic subjects and provides an
additional LDL-cholesterol reduction in individuals on statin therapy.
Journal of Agricultural and Food Chemistry, 57(18), 8316-8324.
Ng, D. S., Chu, T., Esposito, B., Hui, P., Connelly, P. W., & Gross, P. L.
(2008). Paraoxonase-1 deficiency in mice predisposes to vascular
inflammation, oxidative stress, and thrombogenicity in the absence
of hyperlipidemia. Cardiovascular Pathology, 17(4), 226-232. http://
Paganini-Stein, F. L., Schmidt, B., Furlong, E. B., Souza-Soares, L. A.,
Soares, M. C., Vaz, M. R., & Muccillo-Baish, A. L. (2005). Vascular
responses to extractable of Ilex paraguariensis in rats fed standard
and high-cholesterol diets. Biological Research for Nursing, 7(2), 146-
156. http://dx.doi.org/10.1177/1099800405280521. PMid:16267376.
Pang, J., Choi, Y., & Park, T. (2008). Ilex paraguariensis extract ameliorates
obesity induced by high fat diet: role of AMPK in the visceral adipose