Int. J. Morphol.

,
33(4):1510-1517, 2015.

Both Hepatic Lipogenesis and Beta-Oxidation are

Altered in Offspring of Mothers Fed a High-Fat
Diet in the First Two Generations (F1 and F2)

La Lipogénesis Hepática y la Beta-Oxidación están Alteradas en las Crías de Madres

Alimentadas con una Dieta Alta en Grasas en las Dos Primeras Generaciones (F1 y F2)

Wilian Rodrigues Lannes*; Andressa Cabral de Miranda*; Vanessa de Souza-Mello*;

Sandra Barbosa-da-Silva*; Marcia Barbosa Aguila* & Carlos Alberto Mandarim-de-Lacerda*

LANNES, W. R.; MIRANDA, A. C.; SOUZA-MELLO, V.; BARBOSA-DA-SILVA, S.; AGUILA, M. B. & MANDARIM-DE-
LACERDA, C. A. Both hepatic lipogenesis and beta-oxidation are altered in offspring of mothers fed a high-fat diet in the first two
generations (F1 and F2). Int. J. Morphol., 33(4):1510-1517, 2015.

SUMMARY: The high fat (HF) fed mothers may program susceptibility in offspring to chronic diseases and affect subsequent
generations. The present study evaluated the liver structure in adulthood, focusing on the F1 and F2 generations. Females C57BL/6 (F0)
were fed standard chow (SC) or HF diet (8 weeks) prior to mating and during the gestation and lactation to provide the F1 generation
(SC-F1 and HF-F1). All other mothers and offspring fed SC. At 3 months old, F1 females were mated to produce the F2 generation (SC-
F2 and HF-F2). The liver was kept in several fragments and prepared for histological analysis or frozen for biochemical and molecular
analyzes. The F1 and F2 offspring were studied at 3 months old. HF-F1 had higher body mass (BM) compared to SC-F1 (P= 0.001), but
not HF-F2 compared to SC-F2. HF-F1 had glucose intolerance when compared to SC-F1, but not HF-F2 compared to SC-F2. HF-F1 (P=
0.009) and HF-F2 (P= 0.03) showed hyperinsulinemia compared to their counterparts. Both groups HF-F1 and HF-F2 showed more
steatosis than the SC counterparts (F1 and F2, P<0.0001). HF-F1 showed increased expression of PPAR-gamma and SREBP1-c compared
to SC-F1 (P= 0.01). HF-F2 showed increased PPAR-gamma expression compared to SC-F2 (P= 0.04). In conclusion, HF-fed mother
impairs both lipogenesis and beta-oxidation pathways in F1 through upregulation of PPAR-gamma and downregulation of PPAR-alpha.
In F2, the only lipogenesis is enhanced, but it causes a disrupted PPAR balance, favoring the hepatic lipid accumulation and impaired
metabolism in these animals that were not directly exposed to the maternal HF intake.

KEY WORDS: High-fat diet; Hepatic lipogenesis; Beta-oxidation; Maternal obesity; Offspring.

INTRODUCTION

A growing body of evidence has associated maternal beta-oxidation in the liver (Angulo, 2007). Peroxisome
nutrition with altered offspring metabolism (Alfaradhi et al., proliferator-activated receptors (PPARs) are transcription
2014; Ornellas et al., 2015). A high fat (HF) fed mothers factors responsible for the transcription of genes involved
favors the early onset of obesity and its comorbidities in the in hepatic beta-oxidation and lipogenesis (Wahli & Michalik,
offspring even when there is balanced nutrition in their post- 2012). HF-fed mothers cause disrupted PPAR balance in the
weaning life (Gregorio et al., 2010). Hyperinsulinemia seems first generation offspring, favoring the high expression of
to mediate this metabolic disruption as it is able to transpose PPAR-gamma (lipogenic factor) concomitant to decreased
placenta and the liver, as a central organ of metabolism, has expression of PPAR-alpha, essential to mitochondrial beta-
emerged as a potent target to excessive lipids during oxidation (Magliano et al., 2013).
intrauterine life (Yessoufou & Moutairou, 2011).
Hepatic impairments have been described in both
Insulin resistance plays a pivotal role in NAFLD newborn mice (Bringhenti et al., 2015) and in adult mice
genesis as it stimulates lipolysis in white adipose tissue, (Alfaradhi et al.). However, little attention is given to a
augmenting the hepatic input of free fatty acids and impairs possible intergenerational effect of maternal HF diet on liver

*
Laboratory of Morphometry, Metabolism, and Cardiovascular Disease, Biomedical Center, Institute of Biology, State University of Rio de Janeiro, RJ,
Brazil.

1510
LANNES, W. R.; MIRANDA, A. C.; SOUZA-MELLO, V.; BARBOSA-DA-SILVA, S.; AGUILA, M. B. & MANDARIM-DE-LACERDA, C. A. Both hepatic lipogenesis and beta-oxidation are
altered in offspring of mothers fed a high-fat diet in the first two generations (F1 and F2). Int. J. Morphol., 33(4):1510-1517, 2015.

structure and function. The present study aimed to verify Mothers. The body mass (BM) was measured weekly. After
whether the hepatic outcome from pre-gestational and mating, the BM gain was assessed considering the BM before
perinatal maternal obesity persists into the second generation, mating. Two days before mating, the oral glucose tolerance
focusing on the disruption of the main metabolic pathways test was performed (as described below).
related to NAFLD onset.
Offspring

MATERIAL AND METHOD Glucose metabolism. Oral glucose tolerance test (OGTT)
was performed after 6 h fasting. Glucose (1 g/kg) was
administered by orogastric gavage, and blood samples were
Animals and diets. The Animal Ethics Committee of the collected in the caudal vein before and at 15, 30, 60, and 120
State University of Rio de Janeiro approved the study with min after glucose overload. Glucose levels were measured
the protocol no. CEUA/017/2013. All procedures followed using a glucometer (Accu-Chek, Roche Diagnostics,
strictly the guidelines for experimentation with animals Germany). The analysis was based on the area under the curve
(NIH Publication no. 85–23, revised 1996). The animals (AUC) calculated in the interval 0 to 120 min (Prism version
were housed under controlled conditions of temperature 6.05 for Windows, GraphPad software, La Jolla, CA, USA).
(21±2° C) and humidity (60±10%) in a 12-h light/dark
cycle, and had free access to food and water. Body mass and euthanasia. F1 and F2 offspring had their
BM measured weekly and their food and energy intakes
Initially, ten virgin C57BL/6 females with one- measured daily until 3-mo-old, when the animals were food
month-old were divided into two groups according to the deprived for 6 h and anesthetized in the CO2 chamber and
dietary lipid content: SC (standard chow group) and HF blood was collected by cardiac puncture.
(high-fat group). The SC (19% protein, 64% carbohydrate,
17% fat) and the HF diet (19% protein, 32% carbohydrates, Metabolic parameters. Plasma concentrations of insulin
49% fat) were manufactured by PragSolutions (Jau, SP, were measured using enzyme immunoassay (# EZMADP-
Brazil) following the recommendations of the AIN-93G 60K, insulin Cat, Millipore, Missouri kit) with the TP-Reader
(Reeves et al., 1993) and offered since two months before ELX800 (BioTek Instruments, USA). Total cholesterol,
mating until the end of lactation. Those females were the triacylglycerol and alanine aminotransferase (ALT) were
mothers F0. measured using a colorimetric kinetic method (Bioclin System
II, Quibasa, Belo Horizonte, MG, Brazil).
Offspring of the mothers F0 was randomly reduced
to six pups in their litters (with a 1:1 sex ratio). All offspring, Liver structure. Liver fragments obtained from all parts of
no matter what food the mother had access were weaned the organ were prepared for light microscopy. Briefly,
onto the SC diet at postnatal day 21. The groups SC-F1 fragments were fixed in 4% formaldehyde for 48 h at room
and HF-F1 were formed by randomly keeping one male temperature and then embedded in Paraplast plus (Sigma-
per litter (n= 5). Aldrich Co., St. Louis, MO, USA), sectioned at 5 µm of
thickness, and sections were stained with hematoxylin and
Females of the SC-F1 and HF-F1 generations, ever eosin. Digital images were obtained in a BX51 microscope
fed SC, were keep at 3mo old and mated with a male bred (Olympus Co., Tokyo, Japan), and an Infinity 1-5c camera
outside the experiment, without any dietary manipulation, (Lumenera Co., Ottawa, Canada). The volume density of
to produce F2 offspring, and F2 litters followed the same steatosis (Vv [steatosis]) was assessed by point counting (in
protocol as F1 litters until weaning, i.e., all offspring of F1 a 36 points test-system) on at least ten random fields per
and F2 generations were fed SC, but they were labeled animal (Aguila et al., 2003; Catta-Preta et al., 2011).
according to the mother F0:
Hepatic triacylglycerol. Fifty mg of liver was placed in
a) SC-F1: male offspring of the first generation of the SC- Eppendorf containing 1 ml of isopropyl alcohol, and after
F0 mothers; homogenate was macerated and then placed in the sonicator
b) HF-F1: male offspring of the first generation of the HF- (Model LB-130 PB Labormetric, Miami, FL, USA) for 20 s.
F0 mothers; It was subsequently taken to a refrigerated centrifuge at 4 °C
c) SC-F2: male offspring of the second generation of the for 10 min at 4,700 RPM. After this procedure, 5 µL of the
SC-F0 mothers; supernatant was pipetted and mixed with the reagent kit and
d) HF-F2: male offspring of the second generation of the quantified in a spectrophotometer (Bioclin, Quibasa, Belo
HF-F0 mothers. Horizonte, MG, Brazil).
1511
LANNES, W. R.; MIRANDA, A. C.; SOUZA-MELLO, V.; BARBOSA-DA-SILVA, S.; AGUILA, M. B. & MANDARIM-DE-LACERDA, C. A. Both hepatic lipogenesis and beta-oxidation are
altered in offspring of mothers fed a high-fat diet in the first two generations (F1 and F2). Int. J. Morphol., 33(4):1510-1517, 2015.

Western blot analysis. Protein extracts were isolated from P-value <0.05 was considered statistically significant. (Prism
homogenized liver tissues and the total proteins were version 6.05 for Windows, GraphPad software, La Jolla, CA,
evaluated using western blotting, in accordance with the USA).
following steps: Initially, around 100 mg of frozen liver were
homogenized in a buffer containing protease inhibitors. The
homogenates were centrifuged at 4 °C, the supernatants were RESULTS
collected, and the total protein was determined using a BCA
protein assay kit (Thermo Scientific, Rockford, IL, USA). Equal
quantities of total protein were separated using polyacrylamide Mothers
gel electrophoresis. Then, the membrane was blocked by
incubation with 6% (wt/vol) nonfat dry milk in Tris-buffered Body mass. Mothers started the experiment without showing
saline [20 mmol/L Tris/HCl (pH 7.4) and 500 mmol/L NaCl] any significant difference in BM. During the pre-mating period
and sequentially incubated overnight at 4 °C with the following (the first eight weeks of the experiment), the HF mothers
primary antibodies: to analyze the hepatic lipogenesis showed increased BM (+6%) when compared to the SC mothers
(SREBP1-c 68kDa; SC-367) (PPAR-gamma 67kDa, SC-7273). (P= 0.01). However, the mothers of the HF-F1 generation, used
To analyze hepatic beta-oxidation (PPAR-alpha 55 kDa, SC- to produce HF-F2 offspring, have no significant difference in
9000); The membranes were also incubated with anti-b-actin BM during the pre-mating period compared to SC-F1 mothers
(monoclonal antibody; 43 kDa; sc-81178; 1:1000; Santa Cruz (Table I).
Biotechnology). After incubation, the membranes were washed
with TBS-T and incubated with secondary antibodies for 1 h Oral glucose tolerance test- OGTT. OGTT was carried out
at room temperature. The membrane was developed using ECL in the immediate pre-pregnancy period to avoid stress during
Western blotting detection reagents, and images of the blot the pregnancy. The HF-0 group had a higher AUC, showing
were obtained from Bio-Rad image ChemiDoc XRS Molecular impaired glucose tolerance compared to the SC group (P=
Systems (Bio-Rad, Hercules, CA, USA). The 0.0001). However, the HF-F1 mothers showed no significant
chemiluminescent intensity of the bands was quantified using difference in the OGTT during the pre-mating period in
ImageJ software, version 1:49 (NIH imagej.nih.gov/ij, USA). comparison with the SC-F1 mothers (Table I).

Statistical analysis. Data are expressed as mean and the Offspring

associated standard deviation. The sample (n) refers to the
number of litters per group (only one male per litter was kept Body mass. The HF-F1 offspring showed greater BM than the
to form the groups). Data were analyzed using one-way analysis SC-F1 offspring (since weaning until 3-mo-old) (P= 0.001).
of variance followed by the posthoc test of Holm-Sidak. Two- As the animals began to consume SC at weaning, this finding
way ANOVA was performed considering the factors: a) mater- is an effect of maternal nutrition. However, HF-F2 offspring
nal diet (in F0), and b) the offspring generation. In all cases, a showed no significant difference in BM compared to the SC-
F2 offspring (Table I).

Table I. Data presented as means ± standard deviation. Means P<0.05 (†), when compared to the standard chow (SC) group of the
same generation (one way ANOVA and posthoc test of Holm-Sidak).
Data SC-F1 HF-F1 SC-F2 HF-2
Mothers Body mass (g) 20.0±0.2 21.4±0.4 † 19.5±0.3 20.4±0.5
OGTT (AU, auc) 705.5±32.0 990.6±12.5 † 712.1±23.2 730.8±48.1
Offspring Body
Mass (g) 25.2±0.20 28.3±0.50† 25.9±0.18 24.8±0.77
Total cholesterol (mg/dL) 125.3±3.27 172.0±8.81† 111.2±2.69 114.8±1.61
Triacylglycerol (mg/dL) 84.9±1.77 96.8±1.03† 86.8±2.05 88.0±1.66
Insulin (IU/L) 3.1±0.43 5.4±0.52† 3.6±0.33 4.9±0.37†
L iver
Mass (g) 0.5±0.02 0.6±0.01† 0.4±0.02 0.5±0.02
ALT (IU/L) 37.6±0.92 50.6±2.60† 37.4±2.11 37.0±3.39
Triacylglycerol (mg/dL/mg) 1.4±0.11 1.9±0.11† 2.0±0.19 2.3±0.10
Groups: Only original mothers (F0) fed standard chow (SC) or high-fat diet (HF), the subsequent mothers fed standard chow.
Offspring of the first (F1) or second generation (F2) received the label according to their original mothers.
Abbreviations: ALT= alanine aminotransferase; AU= arbitrary units; auc= area under the curve; OGTT= oral glucose tolerance test;
IU= international units.

1512
LANNES, W. R.; MIRANDA, A. C.; SOUZA-MELLO, V.; BARBOSA-DA-SILVA, S.; AGUILA, M. B. & MANDARIM-DE-LACERDA, C. A. Both hepatic lipogenesis and beta-oxidation are
altered in offspring of mothers fed a high-fat diet in the first two generations (F1 and F2). Int. J. Morphol., 33(4):1510-1517, 2015.

Fig. 1. Oral glucose tolerance test. The values are the means and their standard deviations; one way ANOVA
and posthoc test of Holm-Sidak. Groups: Original mothers (F0) fed standard chow (SC) or high-fat diet (HF)
and subsequent mothers fed standard chow. Offspring of the first (F1) or second generation (F2) received the
label according to their original mothers.

Glucose metabolism. The HF-F1 offspring had The OGTT was greater in the HF-F1 offspring
hyperinsulinemia compared to the SC-F1 offspring (P= (+8.6%) compared to the SC-F1 offspring (P= 0.006),
0.009), and the HF-F2 offspring had hyperinsulinemia characterizing glucose intolerance. However, no significant
compared to the SC-F2 offspring (P= 0.03) (Table I). difference in the OGTT was found comparing the offspring
HF-F2 and SC-F2 (Fig. 1).

Fig. 2. Hepatic steatosis. The values are the means and their standard deviations; one way ANOVA and posthoc test of
Holm-Sidak. Groups: Original mothers (F0) fed standard chow (SC) or high-fat diet (HF) and subsequent mothers fed
standard chow. Offspring of the first (F1) or second generation (F2) received the label according to their original mothers.

1513
LANNES, W. R.; MIRANDA, A. C.; SOUZA-MELLO, V.; BARBOSA-DA-SILVA, S.; AGUILA, M. B. & MANDARIM-DE-LACERDA, C. A. Both hepatic lipogenesis and beta-oxidation are
altered in offspring of mothers fed a high-fat diet in the first two generations (F1 and F2). Int. J. Morphol., 33(4):1510-1517, 2015.

Plasma lipids. The HF-F1 offspring showed a significant The volume density of the hepatic steatosis was 180%
increase in triacylglycerol (+14%) and total cholesterol (+37%) greater in the HF-F1 offspring compared to the SC-F1 offspring
levels compared to the SC-F1 offspring (P <0.05). On the other (P<0.0001), and was 86 % greater in the HF-F2 offspring
hand, no difference was seen (total cholesterol and compared to the SC-F2 offspring (P<0.0001) (Fig. 2).
triacylglycerol) comparing the offspring HF-F2 and SC-F2
(Table I). The ALT enzyme, a marker of liver injury, was
elevated in the HF-F1 offspring compared to the SC-F1
Lipogenesis and hepatic steatosis. The mass of the liver was offspring (P= 0.001), but no difference was found comparing
20% greater in the HF-F1 offspring than in the SC-F1 offspring the offspring HF-F2 and SC-F2 (Table I).
(P= 0.006), but the mass of the liver was not different
comparing the groups HF-F2 and SC-F2. Figure 3 shows protein expressions in the liver. In
the first generation, the HF-F1 offspring showed increased
The triacylglycerol content in the liver was 30% higher liver expression of both PPAR-gamma and SREBP1-c
in the HF-F1 offspring than in the SC-F1 offspring (P= 0.03), (associated with lipogenesis) than the SC-F1 offspring (P=
but no difference was found in the triacylglycerol content in 0.01). The HF-F1 offspring showed diminished liver
the liver comparing the offspring HF-F2 and SC-F2 (Table I). expression of PPAR-alpha (associated with beta-oxidation)

Fig. 3. Protein expression in the liver. The values are the means and their standard deviations; one way ANOVA and posthoc
test of Holm-Sidak. Groups: Original mothers (F0) fed standard chow (SC) or high-fat diet (HF) and subsequent mothers fed
standard chow. Offspring of the first (F1) or second generation (F2) received the label according to their original mothers. A)
PPAR-gamma; B) PPAR-alpha; C) SREBP-1c; D) representative bands in the sequence of the groups: SC–F1, HF–F1, SC–
F2 and HF–F2.

1514
LANNES, W. R.; MIRANDA, A. C.; SOUZA-MELLO, V.; BARBOSA-DA-SILVA, S.; AGUILA, M. B. & MANDARIM-DE-LACERDA, C. A. Both hepatic lipogenesis and beta-oxidation are
altered in offspring of mothers fed a high-fat diet in the first two generations (F1 and F2). Int. J. Morphol., 33(4):1510-1517, 2015.

than the SC-F1 offspring (P= 0.04). In the second generation, results of our laboratory as offspring from HF-fed mothers
the HF-F2 offspring showed increased liver expression of were smaller at birth but has a catch-up growth, showing
PPAR-gamma compared to the SC-F2 offspring (P= 0.04). overweight at weaning (Gregorio et al.; Bringhenti et al.;
Although there was no difference regarding both SREBP-1 Graus-Nunes et al., 2015).
and PPAR-alpha. The PPAR-gamma overexpression is
probably associated with high hepatic lipogenesis, causing The maternal factors that cause the metabolic
hepatic steatosis as found in the HF-F2 offspring. disorders in the offspring are unknown and hyperinsulinemia
seems to play a role (Barbosa-da-Silva et al., 2014). The
Two-way ANOVA interactions. There was a decline in glucose homeostasis develops with age in models
significant interaction between maternal diet and offspring of developmental programming. Offspring of obese mothers
generation in the following offspring data: body mass (P= becomes hyperinsulinemic at 3 months-old, progressing to
0.008), plasma triacylglycerol (P= 0.007), total cholesterol type 2 diabetes at 6 months old (Samuelsson et al., 2008). It
(P<0.0001), liver mass (P= 0.005), ALT (P= 0.006) and liver is already described that the offspring of HF-fed mothers
steatosis (P<0.0001). develops insulin resistance (Buckley et al.) and glucose
intolerance in adulthood (Samuelsson et al.). The present
findings confirm the alterations in both insulin and glucose
DISCUSSION metabolisms in the F1 offspring, but not in the F2 offspring.

The HF-fed mothers have been shown to cause

The present study provides new information increased levels of triacylglycerol in the maternal circulation,
concerning the impact of a maternal HF diet during which may enhance the concentration across the placenta,
pregnancy and lactation using a mouse model, focusing on resulting in higher transport and deposition of lipids in fetal
the intergenerational effects on the body mass, glucose organs (Carmody et al., 2011). In fact, increased total
metabolism and hepatic steatosis in the second-generation cholesterol, plasma, and liver triacylglycerol were found in
(F2) offspring. The liver is highly susceptible to the the HF-F1 offspring, but the HF-F2 offspring seems to have
detrimental effects of fetal overnutrition (Nathanielsz & been spared these alterations. Of course, the effects on
Hanson, 2003). Our results show that body mass, OGTT, subsequent generations may differ from those seen in the
insulinemia, plasma lipids and liver data were more altered F1, who were exposed directly to the insult in utero (Drake
in the F1 offspring in comparison with the F2 offspring. et al., 2011; Dunn & Bale). The maternal insulin resistance
and HF diet combined showed additional effects on adiposity
Evidence both from human and animal studies in the offspring early life (Carmody et al.; Barbosa-da-Silva
suggests that fetal programming may not be limited to the et al.).
directly exposed F1 but may be transmitted to subsequent
generations without re-exposure (Jimenez-Chillaron et al., Offspring of obese mothers is fatter and has the risk
2009; Drake & Liu, 2010). These effects may be of developing fatty liver compared with offspring of lean
transmissible through both maternal and paternal lines mothers (Bruce et al., 2009; Elahi et al., 2009). Our study
(Jimenez-Chillaron et al.; Pentinat et al., 2010; Dunn & Bale, demonstrated the presence of microvesicular hepatocyte
2011). steatosis in the offspring of mothers who received the HF
diet in both F1 and F2. Liver and adipose tissue are differently
Murine models are widely used to address fetal affected by HF-fed mothers, leading to upregulated hepatic
programming and maternal HF feeding during pregnancy lipogenesis coupled with blunted lipolysis in white adipose
have caused diverse effects on fetal birth weight, including tissue. Critical points of regulation in the respective pathways
no effect, decreased birth weight, and increased birth weight are associated with a high expression of SREBP-1c and
(Buckley et al., 2005; Howie et al., 2009). The maternal downstream genes involved in hepatic lipid biosynthesis
obesity programs to an increased sensitivity to weight gain concomitant with reduced expression of PPAR-alpha and
in the offspring in the absence of changes in birth weights related genes involved in fatty acid oxidation (Shankar et
(Alfaradhi et al.). We observed the HF-F1 offspring with al., 2010), which agrees with our results. However, the
higher BM during their lives compared to the SC-F1 steatosis observed in F2, probably was favored hepatic
offspring, but not in the F2. Another study of chronic lipogenesis through enhanced PPAR-gamma expression.
administration a HF diet both before and during pregnancy PPAR-gamma downstream effects, including induction of
resulted in decreased fetal birth weight in rodents, which SREBP-1c, are responsible for regulating the genes required
suggests some degree of placental insufficiency (Howie et for de novo lipogenesis and, consequently, increased lipid
al.). The present findings are in agreement with previous accumulation (Knebel et al., 2012).
1515
LANNES, W. R.; MIRANDA, A. C.; SOUZA-MELLO, V.; BARBOSA-DA-SILVA, S.; AGUILA, M. B. & MANDARIM-DE-LACERDA, C. A. Both hepatic lipogenesis and beta-oxidation are
altered in offspring of mothers fed a high-fat diet in the first two generations (F1 and F2). Int. J. Morphol., 33(4):1510-1517, 2015.

In conclusion, HF-fed mothers led to el grupo CS-F1 (P= 0,01). HF-F2 mostró aumento de la expre-
intergenerational effects on liver structure and metabolism, sión de PPAR-gamma en comparación con CS-F2 (P= 0,04). En
more pronounced in F1, but F2 also showed important conclusión, la madre alimentada con HF presenta ambas vías
alterations in the liver structure. We proposed that the HF- afectadas, de lipogénesis y de la beta-oxidación, en la F1 a tra-
vés de la regulación positiva de PPAR-gamma y con regulación
fed mothers impair both lipogenesis and beta-oxidation
a la baja de los PPAR-alfa. En F2, solo ha mejorado la vía de
pathways through upregulation of PPAR-gamma and lipogénesis, pero causa un desbalance de PPAR, lo que favorece
downregulation of PPAR-alpha. The F2 offspring has la acumulación de lípidos hepáticos y la alteración del metabo-
enhanced lipogenesis, but it causes disrupted PPAR balan- lismo en estos animales que no estaban directamente expuestos
ce, favoring hepatic lipid accumulation in these animals that a la ingesta materna de HF.
were not directly exposed to the mother nutrition.
PALABRAS CLAVE: Dieta alta en grasa;
Lipogénesis hepática; Beta oxidación; Obesidad materna;
ACKNOWLEDGMENTS Descendencia.

The authors would like to thank Mrs. Aline Penna,

REFERENCES
Mrs. Gezileia Lau, and Mrs. Michele Soares for their
technical assistance. This study was supported by Conselho
Nacional de Desenvolvimento Científico e Tecnológico Aguila, M. B.; Pinheiro, Ada. R.; Parente, L. B. & Mandarim-de-
(CNPq), Fundação de Amparo a Pesquisa do Estado do Rio Lacerda, C. A. Dietary effect of different high-fat diet on rat
de Janeiro (FAPERJ), and Coordenação de Aperfeiçoamento liver stereology. Liver Int., 23(5):363-70, 2003.
de Pessoal de Nível Superior (CAPES), Brazil.
Alfaradhi, M. Z.; Fernandez-Twinn, D. S.; Martin-Gronert, M. S.;
Musial, B.; Fowden, A. & Ozanne, S. E. Oxidative stress and
altered lipid homeostasis in the programming of offspring fatty
LANNES, W. R.; MIRANDA, A. C.; SOUZA-MELLO, V.; liver by maternal obesity. Am. J. Physiol. Regul. Integr. Comp.
BARBOSA-DA-SILVA, S.; AGUILA, M. B. & MANDARIM- Physiol., 307(1):R26-34, 2014.
DE-LACERDA, C. A. La lipogenesis hepática y la beta-oxida-
ción están alteradas en las crías de las madres alimentadas con Angulo, P. Obesity and nonalcoholic fatty liver disease. Nutr. Rev.,
una dieta alta en grasas en las dos primeras generaciones (F1 y 65(6 Pt. 2):S57-63, 2007.
F2). Int. J. Morphol., 33(4):1510-1517, 2015.
Barbosa-da-Silva, S.; Sarmento, I. B.; Bargut, T. C. L.; Souza-Me-
RESUMEN: Los madres alimentadas con dieta rica en llo, V.; Aguila, M. B. & Mandarim-de-Lacerda, C. A. Animal
grasas (HF) pueden programar una susceptibilidad al desarrollo models of nutritional induction of type 2 diabetes mellitus.
de enfermedades crónicas en su descendencia y de este modo Int. J. Morphol., 32(1):279-93, 2014.
afectar a las generaciones posteriores. El presente estudio eva-
luó la estructura del hígado en la edad adulta, centrándose en las Bringhenti, I.; Ornellas, F.; Martins, M. A.; Mandarim-de-Lacerda,
generaciones F1 y F2. Las hembras C57BL/6 (F0) fueron ali- C. A. & Aguila, M. B. Early hepatic insult in the offspring of
mentadas con dieta estándar (CS) o dieta HF (8 semanas) antes obese maternal mice. Nutr. Res., 35(2):136-45, 2015.
del apareamiento y durante la gestación y lactancia para produ-
cir la generación F1 (CS-F1 y HF-F1). Todas las demás madres Bruce, K. D.; Cagampang, F. R.; Argenton, M.; Zhang, J.; Ethirajan,
y crías fueron alimentadas con CS. A los 3 meses de edad, las P. L.; Burdge, G. C.; Bateman, A. C.; Clough, G. F.; Poston,
hembras F1 fueron apareadas para producir la generación F2 L.; Hanson, M. A.; McConnell, J. M. & Byrne, C. D. Maternal
(CS-F2 y HF-F2). El hígado se conservó en varios fragmentos y high-fat feeding primes steatohepatitis in adult mice offspring,
se preparó, por un lado, para el análisis histológico, y por otro, involving mitochondrial dysfunction and altered lipogenesis
se lo congeló para realizar análisis bioquímicos y moleculares. gene expression. Hepatology, 50(6):1796-808, 2009.
La descendencia F1 y F2 se estudió a los 3 meses de edad. HF-
F1 tuvo una mayor masa corporal (BM) en comparación con Buckley, A. J.; Keserü, B.; Briody, J.; Thompson, M.; Ozanne, S.
CS-F1 (P= 0,001), pero no el grupo HF-F2 en comparación con E. & Thompson, C. H. Altered body composition and
CS-F2. HF-F1 tenía intolerancia a la glucosa en comparación metabolism in the male offspring of high fat-fed rats.
con CS-F1, pero no el grupo HF-F2 en comparación con CS- Metabolism, 54(4):500-7, 2005.
F2. HF-F1 (P= 0,009) y HF-F2 (P= 0,03) mostraron
hiperinsulinemia en comparación con sus homólogos. Ambos Carmody, J. S.; Wan, P.; Accili, D.; Zeltser, L. M. & Leibel, R. L.
grupos HF-F1 y HF-F2 mostraron más esteatosis que las con- Respective contributions of maternal insulin resistance and diet
trapartes CS (F1 y F2, P <0,0001). HF-F1 mostró una mayor to metabolic and hypothalamic phenotypes of progeny. Obesity
expresión de PPAR-gamma y SREBP1-c en comparación con (Silver Spring), 19(3):492-9, 2011.

1516
LANNES, W. R.; MIRANDA, A. C.; SOUZA-MELLO, V.; BARBOSA-DA-SILVA, S.; AGUILA, M. B. & MANDARIM-DE-LACERDA, C. A. Both hepatic lipogenesis and beta-oxidation are
altered in offspring of mothers fed a high-fat diet in the first two generations (F1 and F2). Int. J. Morphol., 33(4):1510-1517, 2015.

Catta-Preta, M.; Mendonca, L. S.; Fraulob-Aquino, J.; Aguila, M. Ornellas, F.; Souza-Mello, V.; Mandarim-de-Lacerda, C. A. & Agui-
B. & Mandarim-de-Lacerda, C. A. A critical analysis of three la, M. B. Programming of obesity and comorbidities in the
quantitative methods of assessment of hepatic steatosis in liver progeny: lessons from a model of diet-induced obese parents.
biopsies. Virchows Arch., 459(5):477-85, 2011. PLoS One, 10(4):e0124737, 2015.

Drake, A. J. & Liu, L. Intergenerational transmission of Pentinat, T.; Ramon-Krauel, M.; Cebria, J.; Diaz, R. & Jimenez-
programmed effects: public health consequences. Trends Chillaron, J. C. Transgenerational inheritance of glucose
Endocrinol. Metab., 21(4):206-13, 2010. intolerance in a mouse model of neonatal overnutrition.
Endocrinology, 151(12):5617-23, 2010.
Drake, A. J.; Liu, L.; Kerrigan, D.; Meehan, R. R. & Seckl, J. R.
Multigenerational programming in the glucocorticoid Reeves, P. G.; Nielsen, F. H. & Fahey, G. C. Jr. AIN-93 purified
programmed rat is associated with generation-specific and diets for laboratory rodents: final report of the American
parent of origin effects. Epigenetics, 6(11):1334-43, 2011. Institute of Nutrition ad hoc writing committee on the
reformulation of the AIN-76A rodent diet. J. Nutr.,
Dunn, G. A. & Bale, T. L. Maternal high-fat diet effects on third- 123(11):1939-51, 1993.
generation female body size via the paternal lineage.
Endocrinology, 152(6):2228-36, 2011. Samuelsson, A. M.; Matthews, P. A.; Argenton, M.; Christie, M.
R.; McConnell, J. M.; Jansen, E. H.; Piersma, A. H.; Ozanne,
Elahi, M. M.; Cagampang, F. R.; Mukhtar, D.; Anthony, F. W.; S. E.; Twinn, D. F.; Remacle, C.; Rowlerson, A.; Poston, L. &
Ohri, S. K. & Hanson, M. A. Long-term maternal high-fat Taylor, P. D. Diet-induced obesity in female mice leads to
feeding from weaning through pregnancy and lactation offspring hyperphagia, adiposity, hypertension, and insulin
predisposes offspring to hypertension, raised plasma lipids resistance: a novel murine model of developmental
and fatty liver in mice. Brit. J. Nutr., 102(4):514-9, 2009. programming. Hypertension, 51(2):383-92, 2008.

Graus-Nunes, F.; Dalla Corte Frantz, E.; Lannes, W. R.; da Silva Shankar, K.; Kang, P.; Harrell, A.; Zhong, Y.; Marecki, J. C.; Ronis,
Menezes, M. C.; Mandarim-de-Lacerda, C. A. & Souza-Me- M. J. & Badger, T. M. Maternal overweight programs insulin
llo, V. Pregestational maternal obesity impairs endocrine and adiponectin signaling in the offspring. Endocrinology,
pancreas in male F1 and F2 progeny. Nutrition, 31(2):380-7, 151(6):2577-89, 2010.
2015.
Wahli, W. & Michalik, L. PPARs at the crossroads of lipid signaling
Gregorio, B. M.; Souza-Mello, V.; Carvalho, J. J.; Mandarim-de- and inflammation. Trends Endocrinol. Metab., 23(7):351-63,
Lacerda, C. A. & Aguila, M. B. Maternal high-fat intake 2012.
predisposes nonalcoholic fatty liver disease in C57BL/6
offspring. Am. J. Obstet. Gynecol., 203(5):495.e1-8, 2010. Yessoufou, A. & Moutairou, K. Maternal diabetes in pregnancy:
Howie, G. J.; Sloboda, D. M.; Kamal, T. & Vickers, M. H. Mater- early and long-term outcomes on the offspring and the concept
nal nutritional history predicts obesity in adult offspring of "metabolic memory". Exp. Diabetes Res., 2011:218598,
independent of postnatal diet. J. Physiol., 587(Pt. 4):905-15, 2011.
2009.

Jimenez-Chillaron, J. C.; Isganaitis, E.; Charalambous, M.; Gesta, Correspondence to:

S.; Pentinat-Pelegrin, T.; Faucette, R. R.; Otis, J. P.; Chow, A.; Sandra Barbosa-da-Silva
Diaz, R.; Ferguson-Smith, A. & Patti, M. E. Intergenerational Laboratório de Morfometria e Morfologia Cardiovascular
transmission of glucose intolerance and obesity by in utero Centro Biomédico
undernutrition in mice. Diabetes, 58(2):460-8, 2009. Instituto de Biologia
Universidade do Estado do Rio de Janeiro
Knebel, B.; Haas, J.; Hartwig, S.; Jacob, S.; Köllmer, C.; Nitzgen, AV. 28 de Setembro 87 (fds) 20551-030
U.; Muller-Wieland, D. & Kotzka, J. Liver-specific expression Rio de Janeiro, RJ
of transcriptionally active SREBP-1c is associated with fatty BRAZIL
liver and increased visceral fat mass. PloS One, 7(2):e31812,
2012.
Email: [email protected] www.lmmc.uerj.br
Magliano, D. C.; Bargut, T. C.; de Carvalho, S. N.; Aguila, M. B.;
Mandarim-de-Lacerda, C. A. & Souza-Mello, V. Peroxisome
proliferator-activated receptors-alpha and gamma are targets to Received: 07-07-2015
treat offspring from maternal diet-induced obesity in mice. PLoS Accepted: 08-09-2015
One, 8(5):e64258, 2013.

Nathanielsz, P. W. & Hanson, M. A. The fetal dilemma: spare the

brain and spoil the liver. J. Physiol., 548(Pt. 2):333, 2003.
1517