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Ilex paraguariensis extract prevents body weight gain in rats fed a high-fat diet


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Studies have shown that drinks containing Ilex paraguariensis extract can promote many benefits in animals and in humans. The present study aimed to evaluate in vivo effects of Ilex paraguariensis extract on metabolic profile, 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. The 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. The 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.
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Food Sci. Technol, Campinas, Ahead of Print, 2019 1/7 1
Food Science and Technology
ISSN 0101-2061 (Print)
ISSN 1678-457X (Online)
1 Introduction
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-inammatory and diuretic
properties (Przygoddaetal., 2010). Likewise, the extract from
Ilex paraguariensis contain dierent 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
eects of the extract (Kangetal., 2012).
Obesity is an increasing problem worldwide, resulting
in signicant morbidity and mortality, as well as a reduced
quality of life (Hurtetal., 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 shis 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 (Menteetal., 2009). Also, it was reported
that consumption of high-fat diets is associated with a reduction
of serum paraoxonase 1 (Pon1) activity (Garciaetal., 2016).
e enzyme Pon1 is a natural antioxidant, and has a primary
role in protecting HDL and LDL from lipid peroxidation (Ngetal.,
2008). Pon1 is synthesized in the liver and secreted into the
bloodstream bound to HDL (Sheetal., 2012). In addition to Pon1,
apolipoprotein A1 (Apoa1) has a specic role in fat metabolism
(Jaichanderetal., 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 (Kontushetal., 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:
Studies have shown that drinks containing Ilex paraguariensis extract can promote many benets in animals and in humans.
e present study aimed to evaluate in vivo eects of Ilex paraguariensis extract on metabolic prole, 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 eect of the high-fat diets.
us, to evaluate the ecacy of potential compounds in the
prevention and treatment of obesity, several animal models have
been used (Kangetal., 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 (Bulloetal., 2007).
Based on these evidences, this study aimed to evaluate the eects
of Ilex paraguariensis extract on metabolic prole, 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. Aer 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
(Reevesetal., 1993) (Table1). 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 aer 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, aer 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
until analysis.
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 coecients of variation found between 11.4% and 17.0%.
e Pon1 activity was measured by its arylesterase activity as
previously established (Sheetal., 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
of CaCl
and 4 mM phenylacetate as substrate. e samples
were diluted 1:3 in a 20 mM Tris/HCl buer 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 coecient. e intra assay coecients 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 Jaé 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 aer 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 puried 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 (Reevesetal., 1993).
Standard Diet High-fat Diet -1 -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 (Table2).
PCR reactions were performed in duplicate in a volume of
12 µL using SYBR Green Mastermix (Applied Biosystems) and the
uorescence was quantied 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 amplication of a single PCR product. Data is
reported as folds over the minimum according to Masternaketal.
(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% signicance level for comparison of
means, using Graphpad Prism 5.0 (GraphPad, La Jolla, CA, USA).
e eects of the diet, supplementation with Ilex paraguaiensis
extract and their interaction were tested. When the interaction
was signicant individual groups were compared by t-test.
3 Results
e results from feed intake indicate that the groups fed
high-fat diet had lower intake (p < 0.01, Table3) when compared
to the standard diet groups, although there was no eect of
Ilex paraguariensis extract supplementation nor interaction
between the diet and treatment (p > 0.05, Table3). Despite no
dierence in feed intake when submitted to the same diet, rats
supplemented with Ilex paraguariensis extract presented smaller
weight gain (p < 0.01). No eect of diet or interaction between Ilex
paraguariensis extract and diet was observed (p > 0.05, Table3).
Ilex paraguariensis extract supplementation was eective to
reduce plasma triglycerides on groups submitted to both diets
(standard or high-fat diet) (p < 0.05, Table4). 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
Pon1 (forward) Pon1 CAAGAACCATCGGTCTTCCT 197
Apoa1 (forward) Apoa1 CAAGAACCATCGGTCTTCCT 828
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: coecient variation; **Average values
followed by the same letter in the column show no signicant statistical dierence 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: coecient variation; **not signicant; ***Average values
followed by the same letter show no signicant statistical dierence 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 signicant changes induced by diet, Ilex paraguariensis
or its interaction (p > 0.01, Table4). Likewise, Pon1 activity
was not aected by diet, Ilex paraguariensis or its interaction
(p > 0.05, Table4).
ere was no dierence in transaminase enzymes GOT
(Figure1A) and GPT (Figure1B) activity between groups
(p > 0.05), indicating no change in liver function for groups
treated with Ilex paraguariensis extract.
Creatinine levels (Figure1C) 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 dierence
(p>0.05) was observed between the groups treated with water
or Ilex paraguariensis extract.
Regarding liver gene expression, Apoa1 (Figure2A), Pon1
(Figure2B), Foxo3 (Figure2E) and Pgc1 (Figure2F) expression
were not dierent between groups. However, Fasn (Figure2C)
and Scd1 (Figure2D) were higher (p < 0.01) in the high-fat + Ilex
paraguariensis group.
4 Discussion
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 eect,
which is reected in the reduced levels of the appetite-stimulating
peptide ghrelin. Thomàs-Moyàetal. (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-Moyàetal., 2007). e reduced intake in rats
fed a high-fat diet is well known, and Haririetal. (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 eect 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 eect
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
Panget al. (2008), indicating that dietary supplementation
with Ilex paraguariensis extract administered to obese rats
induced by high-fat diet was able to signicantly reduce body
weight gain, plasma triacylglycerides, glucose,, along with
reduction on anti-inammatory markers (Luzet al., 2016),
and antidepressant-like eects (Reisetal., 2014). According
to Hetzleretal. (1990) the amount of caeine present in Ilex
paraguariensis could be responsible for the signicant decrease in
the amount of epididymal and abdominal fat. e caeine 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 (Silvaetal., 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 eect (Resendeetal., 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. Silvaetal.
(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 eect, resulting in increased mobilization of fatty
acids from adipose tissue or intramuscular fat depots. However,
Paganini-Steinetal. (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 (Kangetal., 2012). erefore, in
addition to reducing body weight gain, Ilex paraguariensis results
in decreased triglycerides levels which can be benecial in the
prevention of heart diseases.
e present study indicated that total cholesterol, HDL
and Pon1 activity were not aected by diet, Ilex paraguariensis
extract or its interaction. Recently, Bravoetal. (2014) studying
the eect 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 eect on serum lipids or antioxidant status.
Despite that in the hypercholesterolemic rats, Ilex paraguariensis
treatment also had no eect 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 aer consumption of a
high cholesterol diet, pointing out that the potential benec
eect of Ilex paraguariensis on risk factors for cardiovascular
diseases seems to be restricted to already hyperlipidemic
animals. Paganini-Steinetal. (2005) were the rst to report a
signicant reduction in serum total cholesterol and triglycerides
of cholesterol-fed rats aer administration of Ilex paraguariensis
aqueous extract. Thomàs-Moyàetal. (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 caeine in the mate
infusion (Moraisetal., 2009).
e present study also analyzed the GOT and GPT enzymes,
which were not dierent 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
signicant errors, since early changes in glomerular ltration
rate can be “hidden” in normal’s creatinine values (Pintoetal.,
2004). erefore, higher creatinine level can reveal that renal
function is disturbed, indicating that Ilex paraguariensis can be
benecial for kidney function.
e eects of Ilex paraguariensis extract on the gene
expression of antioxidant/inammatory markers have been
studied in animal models (Matsumotoetal., 2009; Arçarietal.,
2011). e analysis of liver tissue expression of Pon1 and
Apoa1, indicated no dierence between diets and treatments.
Similar results were demonstrated by Boaventuraetal. (2012)
in humans, indicating that there was no change in Pon1 activity
aer a prolonged ingestion (90 days) of Ilex paraguariensis tea.
However, Meninietal. (2007) reported increases in serum Pon1
activity aer 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 Moraisetal. (2009) in humans corroborate
with the data in this study, and also did not observe signicant
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 (Lyssenkoetal., 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 (Yangetal., 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
(Hortonet 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 eect 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 dierent among groups in this
study. Additionally, Pon1 and Apoa1 gene expression were also
not dierent, conrming the observations that serum levels of
Pon1 were not changed as well as of HDL.
5 Conclusion
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
its comorbidities.
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
Cientíco e Tecnológico – Brasil (CNPq) and Fundação de
Amparo à Pesquisa do Estado do Rio Grande do Sul – Brasil
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... On the other hand, ewes supplemented with YM tend to decrease plasma TC values. A likely mechanism for this YM-lowering ability is to block cholesterol absorption in the small intestine and/or inhibit cholesterol synthesis in the liver (Uecker et al. 2019). In fact, the caffeine present in YM suppresses the intestinal absorption of cholesterol by decreasing its micellar solubility (Kang et al. 2012). ...
... Generally, considering that YM contains all these bioactive compounds, we can hypothesize that a synergistic effect of interaction between the constituents may occur, as well as a dual mechanism leading to the hypocholesterolemic outcome (Uecker et al. 2019). ...
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The hypothesis tested is that the association of soybean grain and yerba mate as an antioxidant source can increase the concentration of polyunsaturated fatty acids (PUFA) with lower oxidation of milk from lactating ewes reared in tropical pastures. Sixteen ewes were randomly distributed in a 2x2 factorial scheme (with or without yerba mate at 110 g / kg of dry matter (DM); with or without soybean grain at 210 g/kg of DM). Yerba mate intake reduced DM intake, and concentrations of ether extract (EE), protein, lactose, defatted dry extract, density, omega-3 concentrations, and the ratio between saturated/monounsaturated fatty acids (FA), but increased the concentration of monounsaturated FA and the sequestering power of free radicals by DPPH without altering the concentrations of conjugated dienes and the concentrations of substances reactive to thiobarbituric acid in milk. Supplementation with soybean grain decreased the birth weight of the lambs and increased the concentrations of conjugated linoleic acid (CLA), PUFA, omega - 6, omega ratio - 6/omega – 3, and total blood cholesterol. The addition of yerba mate did not influence the oxidative profile of milk. Supplementation with soybean grain improved the FA profile of milk, increasing the concentrations of CLA and PUFA. The results suggest that the association of yerba mate and soybean grain helps to improve the fat quality of milk from ewes raised in tropical pastures, as shown by the enhanced antioxidant activity, although it does not help prevent oxidation of milk rich in PUFA.
... In popular medicine, it is recommend to trait arthritis, headache, rheumatism, hemorrhoids, obesity, fatigue, slow digestion, and liver disorders among other health problems (Bastos et al., 2005). Yerba mate has been reported to possess multiple properties, such as vasodilatory effects, reducing cholesterol and triglyceride (Stein et al., 2005), preventing DNA damage (Mohadjerani & Roodgar, 2016), antimutagenic and radioprotective effects (Bracesco et al., 2018), antidiabetic effects (Heck & De Mejia, 2007;Rocha et al., 2018), prevention of the senescence of retinal cells and mobilization of the cellular pathways (Tate et al., 2020), antioxidant and antimicrobial effects (Kungel et al., 2018), preservation of the cardiovascular health (Cardozo Junior & Morand, 2016), anti-obesity effects (Gambero & Ribeiro, 2015;Kim et al., 2015;Uecker et al., 2019), prevention of liver redox imbalance, high triglycerides and microsteatosis (de Oliveira et al., 2018), among others. It is also an excellent source of polyphenols such as caffeic and chlorogenic acids (Tate et al., 2020). ...
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Yerba mate (Ilex paraguariensis) is a widely consumed herb which originated in Paraguay but grows widely in the region between Southern Brazil, Northern Argentina, Paraguay, and Uruguay. The plant is reported to possess multiple medicinal properties which makes yerba mate popular for consumption as tea in this region. It is also being considered as a nutraceutical product and a functional food consumed in different countries around the world. In popular medicine, and it has been scientifically reported to possess multiple properties. Thirty-nine brands of yerba mate samples were analyzed by serial dilution plates. and incubated at 25 ° C for 5 days. Based on their morphological characteristics, the fungal incidence and the number of colonies forming units were determined. Of the 39 samples analyzed, 15.4% were free from the presence of fungi and yeasts. The remaining 84.6% of the samples were contaminated with variable contents of the fungi Aspergillus. The presence of Aspergillus sections nigri, flavi, circumdati, fumigati, wentii, usti, versicolor and Emericella nidulans was observed the most prevalent among them was Aspergillus section nigri, representing 65.3%. Considering the health risks associated with these fungi, it is important to consider formulating regulations regarding the particle size and the presence of ochratoxins and aflatoxins in commercial yerba mate products.
The prevalence of obesity is growing worldwide and has been extensively linked to gut microbiota dysbiosis. In addition to exercise and physical activity, fiber-rich foods may be a first-line prophylactic to manage obesity. This study investigated in vivo dietary intervention with high-amylose maize starch (HAMS) and starch-entrapped microspheres (MS) to treat high-fat diet induced metabolic disorder and gut microbiome dysbiosis in mice. MS more efficiently controlled body weight as well as adipose tissue mass compared to HAMS. Furthermore, MS significantly reduced blood glucose, insulin, lipid and pro-inflammatory cytokine levels compared to the high-fat diet, while the effects of HAMS were less pronounced. The MS-altered gut microbiota composition favoring Streptococcaceae, Bacilli, Firmicutes and unclassified Clostridiales was predicted to promote fatty acid, pantothenate and Coenzyme A biosynthesis. In line with this, elevated fecal short chain fatty acid (SCFA), in particular, propionate concentration was observed in MS-fed mice. Our study provides novel insights into the mechanistic action of MS on intestinal homeostasis, providing a basis for future dietary therapeutic applications.
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It has been estimated that more than 70% of all drugs approved worldwide between 1981 and 2006 for human health are derived from or structurally similar to natural compounds. The identification of biological matrices containing bioactive compounds with therapeutic and nutraceutical potential is necessary to supply the global market demands. Researches have indicated that the consumption of dry and aqueous extracts of Ilex paraguariensis A. St.‐Hil. is safe, providing that plant biomass does not be exposed to smoke over the drying process, avoiding contamination (e.g., ) with polycyclic aromatic hydrocarbon compounds, and can might help avoiding many diseases, with important potential applications in the pharma and nutraceutical industries. A survey was carried out covering the main therapeutic and nutraceutical studies performed on I. paraguariensis extracts and their relationship with the global patents granted in the last 20 years for the products using this specie in their composition. In the PubMed database, by searching for the term “Ilex paraguariensis,” an output with 497 scientific publications was found. Each paper was analyzed individually and 26 publications encompassing exclusively therapeutical and nutraceutical approaches of that plant species were selected. For the patent screening regarding Ilex‐derived products, the survey considered three patent databases: European Patent Office (EPO) (Espacenet), World Intellectual Property Organization, WIPO), and National Institute of Industrial Property (NIIP—Brazil). The criterion chosen to select the patents in the databases was the inclusion of the terms “Ilex paraguariensis” and “yerba mate” in the title and/or in the abstract, considering the patents issued from 2000 to 2020. Additionally, only patents with therapeutic and nutraceutical potential were considered on the survey. The screening and selection of the documents were performed independently by two researchers and the information cross‐checked at the end. This review contributes to show the state of the art over the last 20 years on the knowledge about the therapeutical and nutraceutical usages of the yerba mate, associated to a certain number of issued patents. The patent survey afforded 62 relevant documents covering products based on Ilex paraguariensis biomass. Considering the number of patents issued, most of them are related to the pharmaceutical area (30), followed by food supplements and beverages (17), cosmetics (10) and, finally, nutraceuticals (5). A detailed analysis of the patents issued showed that most are related to pharmaceutical grade products, generally, marketed as oral and injectable compositions for treatments of obesity, insulin resistance, hyperlipemia and diabetes mellitus, arteriosclerosis, neurological diseases, and SARS‐Cov‐2, for example. In this work, a curious fact is that there are few patents for food, cosmetics, and nutraceuticals products containing yerba mate. Therefore, it seems to be relevant to take into account the potential of that species as source of bioactive compounds for the development of new products not only intended to the pharma sector. In this sense, 26 reports were identified showing possibilities and trendiness in developing new yerba mate based products, such as packaging, biopesticides, antiseptics, and food supply, expanding the possibilities of technological applications of this plant species.
Ilex paraguariensis St. Hil., from the family of holy plants, Aquifoliaceae, is a native South American tree that can reach 18 m in height and is used to produce “yerba mate”. It is found primarily in the southern regions of South America, namely, Brazil (Mato Grosso do Sul, Minas Gerais, Parana, Rio Grande do Sul, Rio de Janeiro, Santa Catarina, Sao Paulo), Argentina (Corrientes, Misiones), Paraguay (Alto Paraná, Amambay, Caaguazú, Canendiyú, Central, Guaira, Itapuá, Misiones, San Pedro), and Uruguay. “Yerba mate” tea, an infusion made from the leaves of this tree, is a widely consumed nonalcoholic beverage in South America which is gaining rapid introduction into the world market, either as tea itself or as ingredient in formulated foods or dietary supplements. The indigenous people have used it for centuries as a social and medicinal beverage. The container in which it is generally served is a gourd made from the dry and hollowed fruit of Lagenaria vulgaris Ser. (Cucurbitaceae), whose dimensions vary according the region and uses. It is called “mate” in the region. “Mate” has a very important social role. The act of offering and sharing it has connotations like those of the tea ceremony for some Oriental cultures.
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The unique chemical composition of yerba mate and its functionalities suggest that it needs to be explored for its innovation potential. New uses may boost consumption, surpassing the traditional consumption barrier, and making yerba mate accessible on a global level. Thus, to highlight the importance of yerba mate as a potential source of agro-economic resources, we present a review on its botanical, ecological, agronomic, and industrial aspects, along with information on the biochemical composition of this species and its biological activity.
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The prevalence of obesity has increased worldwide over the past three decades. Global anti-obesity strategies focus on dietary and lifestyle modifications to slow the development of obesity. Research in the nutrition field has recently aroused considerable interest based on the potential of natural products to counteract obesity. Several studies have identified yerba maté (Ilex paraguariensis) as an excellent candidate. In this review, we evaluated the impact of yerba maté on obesity and obesity-related inflammation. Cellular studies demonstrate that yerba maté suppresses adipocyte differentiation and triglyceride accumulation and reduces inflammation. Animal studies show that yerba maté modulates signaling pathways that regulate adipogenesis, antioxidant, anti-inflammatory and insulin signaling responses. In summary, the data presented here showed that the use of yerba maté might be useful against obesity, improving the lipid parameters in humans and animal models. In addition, yerba maté modulates the expression of genes that are changed in the obese state and restores them to more normal levels of expression. In doing so, it addresses several of the abnormal and disease-causing factors associated with obesity. Protective and ameliorative effects on insulin resistance were also observed. Thus, as a general conclusion, it seems that yerba maté beverages and supplements might be helpful in the battle against obesity.
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Globally obesity has reached to epidemic proportions, and the people of the Gulf countries have also affected, especially high-income, oil-producing countries. The prevalence of obesity in Gulf Countries among children and adolescents ranges from 5% to 14% in males and from 3% to 18% in females. In adult females there is a significant increase of obesity with a prevalence of 2%-55% and in adult males 1%-30% in countries of gulf region. Over the last two decades there is increased consumption of fast foods and sugar-dense beverages (e.g., sodas). Simultaneously, technological advances - cars, elevators, escalators, and remotes have lead to a decrease in level of activity. Traditional dependence on locally grown natural products such as dates, vegetables, wheat and has also shifted. Changes in food consumption, socioeconomic and demographic factors, physical activity, and urbanization are being important factors that contribute to the increased prevalence of obesity in the region.
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In this study, we investigated the possible antidepressant-like effect of I. paraguariensis in rats. Rats were treated for four weeks with an aqueous extract of I. paraguariensis in drinking water, following the traditional preparation of this beverage. After the period of treatment, behavioral (elevated plus-maze, open field test, and forced swimming test) and biochemical parameters (lipid peroxidation assay, thiol content, vitamin C levels, and monoamine oxidase activity) were evaluated. Animals were also analyzed on forced swimming test after 24 hours of I. paraguariensis intake. An additional group was injected with selegiline 24 hours and 30 minutes before forced swimming test as positive control. HPLC analysis revealed the profile of I. paraguariensis extract. I. paraguariensis reduced the immobility time on forced swimming test without significant changes in locomotor activity in the open field test. Any anxiolytic/anxiogenic effect of I. paraguariensis was observed in rats through the elevated plus-maze test. The antidepressant-like effect of I. paraguariensis was not accompanied by inhibitory effect on monoamine oxidase activity. There were no significant alterations on lipid peroxidation, thiol content, and vitamin C levels among the groups. In conclusion, aqueous extract of I. paraguariensis decreases the time of immobility in rats suggesting an antidepressant-like effect.
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Ilex paraguariensis A. St.-Hil., Aquifoliaceae, is a species native to the subtropical and temperate regions of South America, used in beverages prepared by infusion such as teas, chimarrão and tererê. To investigate the physiological effects of I. paraguariensis on the metabolism of fats and sugars in Wistar rats, following the ingestion of erva-mate tea, four experimental groups were constructed: Lipid Control Group (receiving water and high-fat diet); Lipid Tea Group (extract of I. paraguariensis and high-fat diet); the Sugar Control Group (water and high-sugar diet); and Sugar Tea Group (extract of I. paraguariensis and high-sugar diet). The animals received their particular diet for 60 days, and were weighed weekly. After this period, the plasma concentrations of cholesterol, glucose and triacylglycerides were determined, together with the weight of visceral fat. The data were subjected to statistical analysis with a significance level of p<0.05. The results show that the ingestion of erva-mate affected body weight, visceral fat and plasma glucose, cholesterol and triacylglyceride levels.
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Plasma high-density lipoproteins (HDLs) are small, dense, protein-rich particles as compared to other lipoprotein classes; roughly half of total HDL mass is accounted for by lipid components. Phospholipids predominate in the HDL lipidome, accounting for 20 to 30% of total HDL mass, with lesser proportions of cholesteryl esters (14-18 wt%), triglycerides (3-6 wt%) and free cholesterol (3-5 wt%). Lipidomic approaches have provided initial insights into the HDL lipidome with identification of >200 individual molecular lipids species in normolipidemic HDL. Plasma HDL particles however reveal high levels of structural, compositional and functional heterogeneity. Establishing direct relationships between HDL structure, composition and atheroprotective functions bears the potential to identify clinically relevant HDL subpopulations. Furthermore, development of HDL-based therapies designed to target beneficial subspecies within the circulating HDL pool can be facilitated using this approach. HDL lipidomics can equally contribute to the identification of biomarkers of both normal and deficient HDL functionality, which may prove useful as biomarkers of cardiovascular risk. Numerous technical issues remain however to be addressed in order to make such developments possible. With all technical questions resolved, quantitative analysis of the molecular components of the HDL lipidome will contribute to expand our knowledge of cardiovascular and metabolic diseases.
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Nascent high-density lipoprotein (HDL) particles arise in different sizes. We have sought to uncover factors that control this size heterogeneity. Gel filtration, native PAGE, and protein cross-linking were used to analyze the size heterogeneity of nascent HDL produced by BHK-ABCA1, RAW 264.7, J774, and HepG2 cells under different levels of two factors considered as a ratio, the availability of apolipoprotein AI (apoAI) -accessible cell lipid, and concentration of extracellular lipid-free apoAI. Increases in the available cell lipid:apoAI ratio due to either elevated ATP-binding cassette transporter A1 (ABCA1) expression and activity or raised cell density (i.e., increasing numerator) shifted the production of nascent HDL from smaller particles with fewer apoAI molecules per particle and fewer molecules of choline-phospholipid and cholesterol per apoAI molecule to larger particles that contained more apoAI and more lipid per molecule of apoAI. A further shift to larger particles was observed in BHK-ABCA1 cells when the available cell lipid:apoAI ratio was raised still higher by decreasing the apoAI concentration (i.e., the denominator). These changes in nascent HDL biogenesis were reminiscent of the transition that occurs in the size composition of reconstituted HDL in response to an increasing initial lipid:apoAI molar ratio. Thus, the ratio of available cell lipid:apoAI is a fundamental cause of nascent HDL size heterogeneity, and rHDL formation is a good model of nascent HDL biogenesis.-Lyssenko, N. N., Nickel, M., Tang, C., Phillips, M. C. Factors controlling nascent high-density lipoprotein particle heterogeneity: ATP-binding cassette transporter A1 activity and cell lipid and apolipoprotein AI availability.
The aim of this study was to compare serum lipid profiles and ovarian gene expression between aged and younger female mice fed a control or a high-fat diet for 2 months. For this 16 female mice (C57BL/6) of 4 months (Young, n = 8) or 13 months (Old, n = 8) of age were used. The females were divided into four groups: (i) young females fed a normal diet; (ii) young females fed a high-fat diet; (iii) old females fed a normal diet; and (iv) old females fed a high-fat diet. Food intake was reduced (P < 0.05) in mice fed with a high-fat (2.9 ± 0.1 g) diet in comparison with control mice (3.9 ± 0.1 g). Body weight was higher for old females on the high-fat diet (35.1 ± 0.3 g) than for young females on the same diet (23.3 ± 0.4 g; P < 0.05). PON1 activity was lower in the high-fat than control diet group (114.3 ± 5.8 vs. 78.1 ± 6.0 kU/L, respectively) and was higher in older than younger females (85.9 ± 6.4 vs. 106.5 ± 5.3; P < 0.05, respectively). Females fed a high-fat diet had lower expression of Igf1 mRNA (P = 0.04). There was an interaction between age and diet for the expression of Gdf9 and Survivin, with lower expression in older females in both diets and young females that received the high-fat diet (P < 0.05). Concluding, the high-fat diet reduced the expression of ovarian Igf1 mRNA, and Gdf9 and Survivin mRNA in younger females, which can indicate lower fertility rates. High-density lipoprotein concentration and PON1 activity were higher in aged female mice.
To study the effect of mate (Ilex paraguariensis) on serum lipids and antioxidant status in normocholesterolaemic and hypercholesterolaemic rats. Triglycerides (TG), total, LDL- and HDL-cholesterol levels, total antioxidant capacity (FRAP and ABTS assays), malondialdehyde (MDA) and protein carbonyls were analysed in serum, and MDA, glutathione and antioxidant enzyme activity in livers of rats drinking water or mate fed normal or cholesterol-cholic supplemented diets. ABTS, glutathione and antioxidant enzymes were not affected by any treatment. In normocholesterolaemic animals, mate had no effect on serum lipids or antioxidant status, yet it increased serum carbonyls and liver MDA concentrations. In hypercholesterolaemic rats, mate consumption had no effect on HDL-cholesterol or protein carbonyls, yet it showed a marked hypolipidaemic action, decreasing TG, total and LDL-cholesterol, and serum MDA levels that had been increased after consuming the high-cholesterol diet. Potential beneficial effect of mate on markers of cardiovascular risk seems to be restricted to hyperlipaemic animals.
Ethnopharmacological relevance: Ilex paraguariensis A. St. Hilaire (mate) has traditionally been used in several South American countries to prepare tea-like beverages having stimulant effects on the CNS and appetite. In recent years, however, mate preparations have been recommended putatively as an appetite suppressant and slimming remedy. Moreover, studies carried out on either normal or diet-induced obese rats treated with mate extracts revealed anti-obesity and satiety effects, thus refuting ethnopharmacological data. In this work, the effect of mate on the intra-abdominal and epididymal fat, and glucose oxidation levels after oral administration in male Wistar rats, was studied using crude extract from leaves, unripe fruits, and a chemically well-defined purified saponin fraction (MSF). Material and methods: Saponin, polyphenol and methylxanthine contents in MSF were analyzed by HPLC-PDA and UPLC/Q-TOF-MS. Crude extracts from mate leaves (LAE) and unripe fruits (FHE) were assayed for comparison purposes. Male Wistar rats fed with standard diet and water ad libitum were used as the control group. Results: The fat weight and both liver and adipose glucose oxidation were reduced significantly by MSF (35, 90 and 60%, respectively), while LAE and FHE were less active. Also, a significant lowering of the blood triglycerides level was observed in rats treated with MSF and LAE. All creatinine, urea, and transaminase plasma levels remained unaffected no matter what mate preparation was considered. It is also worth pointing out that the glucose blood level was increased after treatment with FHE. This finding did not correlate either with the content of methylxanthines, polyphenols or saponins. Conclusion: A reduction in both visceral fat weight and glucose oxidation of hepatic and adipose tissue in healthy rats fed with a standard diet could be ascribed to a purified mate saponin fraction from unripe fruits. These findings agree with former studies carried out with crude mate extracts and also suggest their potential use as an anti-obesity preparation. Nonetheless, further in vivo experiments are still required to corroborate its effect on human beings.