Article

Fetal and neonatal exposure to nicotine in Wistar rats results in increased beta cell apoptosis at birth and postnatal endocrine and metabolic changes associated with type 2 diabetes

Department of Obstetrics and Gynaecology, McMaster University, Hamilton, Ontario, Canada
Diabetologia (Impact Factor: 6.67). 01/2006; 48(12):2661-6. DOI: 10.1007/s00125-005-0022-5
Source: PubMed
ABSTRACT
Epidemiological studies report an increased risk of obesity and type 2 diabetes in children born to women who smoked during pregnancy. This study examines the effect of fetal and neonatal exposure to nicotine, the major addictive component of cigarettes, on postnatal growth, adiposity and glucose homeostasis.
Female Wistar rats were given either saline (vehicle) or nicotine (1 mg kg(-1) day(-1)) during pregnancy and lactation. Serum and pancreas tissue were collected from the infant rats at birth. Postnatal growth was assessed weekly until the rats reached 26 weeks of age and glucose homeostasis was examined by OGTTs performed at 7 and 26 weeks of age.
Exposure to nicotine resulted in increased postnatal growth and adiposity. Nicotine exposure also resulted in dysglycaemia at 7 and 26 weeks of age. Serum insulin concentrations were decreased in the pups exposed to nicotine at birth. This was associated with increased beta cell apoptosis (pups of saline-treated mothers 8.8+/-1.21% apoptotic beta cells; pups of nicotine-treated mothers 27.8+/-3.1% apoptotic beta cells).
Fetal and neonatal exposure to nicotine results in metabolic changes in the offspring that are consistent with obesity and type 2 diabetes. We propose that these metabolic changes may be a consequence of the initial insult to the beta cell during fetal life and that this animal model has many characteristics of diabetes in humans.

Full-text

Available from: Katherine M Morrison
Diabetologia (2005) 48: 26612666
DOI 10.1007/s00125-005-0022-5
ARTICLE
A. C. Holloway
.
G. E. Lim
.
J. J. Petrik
.
W. G. Foster
.
K. M. Morrison
.
H. C. Gerstein
Fetal and neonatal exposure to nicotine in Wistar rats results
in increased beta cell apoptosis at birth and postnatal endocrine
and metabolic changes associated with type 2 diabetes
Received: 5 May 2005 / Accepted: 18 July 2005 / Published online: 4 November 2005
# Springer-Verlag 2005
Abstract Aims/hypothesis: Epidemiological studies re-
port an increased risk of obesity and type 2 diabetes in
children born to women who smoked during pregnancy.
This study examines the effect of fetal and neonatal
exposure to nicotine, the major addictive component of
cigarettes, on postnatal growth, adiposity and glucose ho-
meostasis. Methods: Female Wistar rats were given either
saline (vehicle) or nicotine (1 mg kg
1
day
1
) during
pregnancy and lactation. Serum and pancreas tissue were
collected from the infant rats at birth. Postnatal growth
was assessed weekly until the rats reached 26 weeks of age
and glucose homeostasis was examined by OGTTs per-
formed at 7 and 26 weeks of age. Results: Exposure to
nicotine resulted in increased postnatal growth and adi-
posity. Nicotine exposure also resulted in dysglycaemia at
7 and 26 weeks of age. Serum insulin concentrations were
decreased in the pups exposed to nicotine at birth. This
was associated with increased beta cell apoptosis (pups of
saline-treated mothers 8.8±1.21% apoptotic beta cells;
pups of nicotine-treated mothers 27.8±3.1% apoptotic beta
cells). Conclusions/interpretation: Fetal and neonatal ex-
posure to nicotine results in metabolic changes in the
offspring that are consistent with obesity and type 2 dia-
betes. We propose that these metabolic changes may be a
consequence of the initial insult to the beta cell during fetal
life and that this animal model has many characteristics of
diabetes in humans.
Keywords Adult-onset disease
.
Beta cell apoptosis
.
Dyslipidaemia
.
Glucose homeostasis
.
Nicotine
.
Type 2 diabetes
Abbreviations DAPI: 4,6-Diamidino-2-Phenylindole
.
PND1: postnatal day 1
.
PND21: postnatal day 21
.
TUNEL: terminal deoxynucleotidyl transferase mediated
dUTP nick end labelling
Introduction
The intrauterine environment is emerging as an important
determinant of chronic adult diseases including diabetes,
obesity and cardiovascular disease. A variety of physio-
logical stresses during pregnancy are thought to contribute
to the development of adult disease. One such stress is
maternal cigarette smoking. Recent epidemiological stud-
ies have shown a strong relationship between maternal
smoking and subsequent obesity, hypertension and type 2
diabetes in the offspring [18]. Although many women
report a desire to quit smoking while pregnant, nicotine
dependence is a significant element of most patients
smoking behaviour [9]. For this reason nicotine replace-
ment therapy has been widely developed as a pharmaco-
therapy of smoking cessation, and is considered to be of
benefit for pregnant women who are highly dependent and
have been unable to quit smoking by other means [911].
However, very few studies have studied the safety of
nicotine replacement during pregnancy [12].
In animal studies, prenatal exposure to nicotine alone
was sufficient to produce reduced birthweight and in-
creased body weight post-natally [13] and significantly
A. C. Holloway (*)
.
G. E. Lim
.
W. G. Foster
Department of Obstetrics and Gynecology,
McMaster University,
Rm 3N52 HSC 1200 Main Street W.,
Hamilton, ON, Canada L8N 3Z5
e-mail: hollow@mcmaster.ca
Tel.: +1-905-5259140
Fax: +1-905-5242911
J. J. Petrik
Department of Biomedical Sciences, University of Guelph,
Guelph, ON, Canada
K. M. Morrison
Department of Pediatrics, McMaster University,
Hamilton, ON, Canada
H. C. Gerstein
Department of Medicine, McMaster University,
Hamilton, ON, Canada
G. E. Lim
Department of Physiology, University of Toronto,
Toronto, ON, Canada
Page 1
higher levels of body fat in fetuses of nicotine-exposed
dams at gestation day 20 (term 22 days) [14]. The fact that
these are risk factors for future obesity and dysglycaemia
[15] suggests that prenatal nicotine exposure may have
lifelong metabolic effects that begin at birth. The effect of
fetal and neonatal nicotine exposure on growth, weight,
adiposity, glucose homeostasis and beta cell area is re-
ported in this study.
Subjects, materials and methods
Maintenance and treatment of animals
All animal experiments were approved by the Animal
Research Ethics Board at McMaster University, in accor-
dance with the guidelines of the Canadian Council for
Animal Care. Nulliparous 200250-g female Wistar rats
(Harlan, Indianapolis, IN, USA) were maintained under
controlled lighting (12:12h light:dark) and temperature
(22°C) with free access to food and water. Two weeks prior
to mating the dams were randomly assigned to receive
either saline (vehicle) or nicotine (n=4 per group). Dams
were injected s.c. with 1 mg·kg
1
·day
1
nicotine bitartrate
(Sigma Aldrich, St Louis, MO, USA) or saline for 14 days
prior to mating, and during pregnancy until weaning. Pups
were weighed after birth (postnatal day 1; PND1) and the
litters were culled to eight at birth, retaining males in
preference to assure uniformity of litter size between
treated and control litters. Trunk blood was collected from
culled animals and allowed to clot at 4°C, then centrifuged
and stored at 80°C until analysis for insulin and glucose
concentrations. Serum glucose concentrations were mea-
sured using a commercially available kit for the glucose
oxidase method (Pointe Scientific, Canton, MI, USA), and
insulin levels were measured on an ultra-sensitive rat
insulin ELISA (Crystal Chem, Downers Grove, IL, USA).
The insulin ELISA kit had a detection limit of 5 pg/ml with
intra- and interassay variabilities of 5.5 and 4.8%, respec-
tively. After weaning at postnatal day 21 (PND21), male
offspring were selected randomly and caged as sibling
pairs. Pups were weighed weekly from PND1 until 26 weeks
of age. Fasting serum samples were collected at 7, 15 and
26 weeks and were analysed for triglyceride concentrations
using a commercially available assay (Pointe Scientific).
Glucose tolerance tests
Glucose tolerance was serially investigated in nicotine-
exposed and control rats at 7 and 26 weeks (n=16 per
group). Serum concentrations of insulin and glucose were
measured in saphenous vein samples obtained after food
deprivation at 0 (0900 hours), 30 and 120 min after rats
were given 2 g/kg glucose (Sigma-Aldrich) in water by
gavage. Blood samples were allowed to clot at 4°C, then
centrifuged and stored at 80°C until used for measure-
ment of serum glucose and insulin concentrations as de-
scribed above.
Immunohistochemistry: beta cell area
To assess whether nicotine exposure can alter pancreas
development, pancreas tissue was removed from all culled
pups at birth (PND1). The pancreas from each animal was
weighed and then fixed by immersion in 10% neutral-
buffered formalin (EM Science, Gibbstown, NJ, USA) at
4°C overnight, washed in PBS and embedded in paraffin.
Immunohistochemical detection of insulin was performed
on 5-μm sections of neonatal pancreatic tissue (PND1;
n=10 per group). Tissue sections were deparaffinised in
xylene, rehydrated and washed in PBS. After quenching in
methanol of endogenous peroxidase activity, antigen
retrieval was performed in 10 mmol/l citrate buffer (pH
3.0) followed by incubation with 10% normal goat serum
and 1% BSA. Sections were then incubated overnight at
4°C with the primary antibody, a polyclonal, guinea-pig
anti-human insulin antibody (1:150 dilution) (DakoCyto-
mation, Carpinteria, CA, USA) which has been shown by
the manufacturer to cross-react with rat insulin peptides.
Sections were then washed in PBS, and immunostaining
was identified using the Vectastain kit (Vector Labora-
tories, Burlinghame, CA, USA) with diaminobenzadine as
the chromogen. Tissue sections were counterstained,
dehydrated and mounted with Permount (Fisher Scientific,
Fair Lawn, NJ, USA). Control sections were incubated
with 1% BSA in PBS in place of the primary antibody. All
measurements were performed at 10× magnification, and
the whole pancreas was analysed. Immunopositive cells
were identified and the beta cell area was calculated as a
percentage of the total area of the section using integrated
morphometry software (M
ETAMORPH, Universal Imaging,
Downington PA, USA). At least 25 fields were counted per
animal.
Immunohistochemistry: detection of beta cell
apoptosis
Histological sections of pancreas from animals at PND1
were processed and fixed as described above. Apoptotic
islet cells were detected using a terminal deoxynucleotidyl
transferase mediated dUTP nick end labelling (TUNEL) kit
(Roche Applied Science, Laval, PQ, Canada) following
the manufacturers protocol. Briefly, after fixation, tissues
and cells were made permeable in 0.1% Triton X-100 for
15 min, washed in PBS, and incubated with the fluorescein
isothiocyanate-conjugated TUNEL enzyme for 60 min to
detect DNA fragmentation. Nuclei were counterstained
with propidium iodide and images were obtained on an
Olympus BX-61 microscope. For analysis, ten fields of
view at 250× magnification were counted for each animal
(n=5 animals per group). To evaluate the percentage of beta
cells that were apoptotic in the pancreas islets, pancreas
tissue from pups at PND1 was processed for double-label
immunofluorescence. For double-label immunofluores-
cence, sections were initially analysed on an apoptosis
detection kit, as described above. Following staining for
fragmented DNA, sections were incubated overnight at
2662
Page 2
4°C with an anti-insulin antibody (DakoCytomation, Car-
pinteria, CA, USA; 1:150). A Texas Red-conjugated anti-
mouse antibody (Vector Laboratories, Burlington, ON,
USA; 1:100 dilution) was used for detection. Nuclei were
counterstained with propidium iodide or DAPI and images
of the specimens were obtained on an Olympus BX-61
fluorescent microscope.
Statistical analysis
All statistical analyses were performed using SigmaStat
(v.2.03, SPSS, Chicago, IL, USA), and t-test or analyses of
variance (ANOVA) followed by appropriate post-hoc tests
when significance was indicated by the ANOVA (α=0.05).
Data were tested for normality as well as equal variance,
and when normality or variance tests failed, data were
analysed using MannWhitney rank sum test or Kruskal
Wallis one-way ANOVA on ranks. Area under the curve for
the total glucose response following the glucose tolerance
test and postnatal growth was assessed using the trapezoidal
rule.
Results
Birth phenotype
Birthweight was reduced in rats that were exposed to
nicotine in utero (saline 6.54±0.099 g, nicotine 6.29±0.056 g,
n=47 and 59, p<0.05) whereas the litter size, sex ratio and
gestational length were unaffected (data not shown).
Postnatal growth and adiposity
There was no significant effect of treatment on weight gain
during lactation and no significant difference in body
weight at weaning. Following weaning the growth trajectory
of the offspring of the nicotine-treated dams was sig-
nificantly enhanced, resulting in increased body weight at
26 weeks of age compared to control offspring (Fig. 1). We
have previously shown that this weight gain is partly the
result of a significant increase in body fat at 26 weeks in
nicotine-exposed pups [16]. Furthermore, in this study
hypertriglyceridaemia was evident in nicotine-exposed
pups at 7 weeks of age (control pups 0.81±0.119 mmol/l,
nicotine-exposed pups 1.08±0.071 mmol/l, p=0.052, n=16
per group) and persisted until 26 weeks of age (control
pups 1.69±0.063 mmol/l, nicotine-exposed pups 2.00±
0.19 mmol/l, p=0.044, n=16 per group).
Glucose homeostasis
Serum glucose and insulin levels were measured at PND1
and at 7 and 26 weeks of age following an overnight fast
and following an oral glucose challenge at 7 and 26 weeks
of age. Fetal nicotine exposure did not alter basal serum
glucose concentrations at birth but caused a significant
reduction in basal serum insulin concentrations in the
nicotine-exposed offspring at PND1 (Table 1). At 7 weeks
of age, there was no difference in fasting serum glucose or
insulin concentrations; however, nicotine-exposed off-
A
g
e
(
weeks
)
0 5 10 15 20 25 30
0
100
200
300
400
500
600
700
Body weight (g)
Fig. 1 Postnatal growth of the
offspring of dams given saline
(control: closed circles, n=16) or
nicotine bitartrate (1 mg/kg/day;
open circles, n=16) during
pregnancy and lactation. Data
are presented as mean±SEM.
Offspring of nicotine-exposed
dams were significantly heavier
than those of saline-treated
controls at the end of the study
(26 weeks of age, p<0.01). The
total growth response (AUC) of
the pups exposed to nicotine in
utero and during lactation was
significantly (p<0.001) higher
than the saline-exposed group
(AUC control pups: 9,299±
231.4; nicotine-exposed pups:
10,826±121.2)
Table 1 Effect of in utero exposure to nicotine on serum hormone
concentrations, pancreatic beta cell area and islet apoptosis at PND1
PND1 Saline Nicotine p value
Glucose (mmol/l) 0.82±0.094 0.89±0.081 0.59
Insulin (ng/ml) 0.36±0.048 0.23±0.032* 0.04
Beta cell area (% of total) 2.5±0.29 3.0±0.31 0.23
Islet apoptosis (% of total) 4.2±0.09 8.7±0.61* <0.001
Data are presented as mean±SEM
2663
Page 3
spring exhibited dysglycaemia following administration of
the oral glucose challenge with no significant difference in
the insulin response to the glucose challenge between the
two groups of animals (Fig. 2).
At 26 weeks there was no significant difference in basal
fasting serum glucose levels between saline- and nicotine-
exposed offspring, but the fasting insulin concentrations
were significantly elevated in the nicotine-exposed off-
020406080100120
2
4
6
8
10
12
Serum glucose (mmol/l)
Time (minutes)
*
a
Serum insulin (pg/ml)
Time (minutes)
020406080100120
0
100
200
300
400
500
600
700
b
020406080100120
4
6
8
10
12
14
Serum glucose (mmol/l)
Time
(
minutes
)
*
*
c
020406080100120
0
500
1000
1500
2000
2500
3000
3500
Time (minutes)
*
*
*
Serum insulin (pg/ml)
d
Fig. 2 Serum glucose and insulin concentrations following
administration of an oral glucose load (2 g/kg body weight
represented by the arrow) at 7 and 26 weeks of age for the
offspring of dams given saline (control: closed circles, n=16) or
nicotine bitartrate, (1 mg/kg/day; open circles, n=16) during
pregnancy and lactation. Data are presented as mean±SEM.
*p<0.05 for difference from control animals at the same time
point. Serum glucose concentrations at 7 weeks of age (a). Nicotine-
exposed rats had higher blood glucose levels at 120 min post-
challenge (p<0.01) than controls. Although the serum glucose
concentration in nicotine-exposed offspring was higher at 30-min
post-challenge, this did not reach significance (p=0.07). The total
glucose response (AUC) following glucose administration was
significantly elevated in the nicotine-exposed offspring (AUC
control pups 12,621±762.9; AUC nicotine-exposed pups 16,376±
985.5, p=0.006) compared to saline controls. b Serum insulin
concentrations at 7 weeks of age. There was no significant
difference at any time point in the total insulin response (AUC)
between the two groups. c Serum glucose concentrations at
26 weeks of age. Nicotine-exposed rats had higher (p=0.001) peak
glucose concentrations at 30 min. Although the serum glucose
concentrations at 120-min post-challenge were higher than those of
the saline controls, this did not reach significance (p=0.07). The total
glucose response to the challenge (AUC) was higher in the nicotine-
exposed animals than in saline controls (AUC control rats 17,341±
1073.5, n=16; AUC nicotine-exposed rats 21,890±927.8, n=16;
p=0.004). d Serum insulin concentrations following an OGTT at
26 weeks of age. Nicotine-exposed offspring had higher serum
insulin concentrations at all time points (p<0.01 for all). The total
insulin response to the glucose challenge was higher in the offspring
of nicotine-treated dams (AUC control rats 77.3±10.5; AUC
nicotine-exposed rats 250.3±42.6; p<0.001)
2664
Page 4
spring. Nicotine-exposed rats exhibited impaired glucose
homeostasis and a significantly higher insulin response to
the glucose challenge (Fig. 2).
Beta cell area and apoptosis
To assess whether the differences in serum insulin levels at
PND1 were a result of an altered number of beta cells or
increased beta cell death, we evaluated both the beta cell
area and the percentage of apoptotic beta cells in the
pancreas at PND1. There was no significant effect of
treatment on beta cell area; however, the offspring of the
nicotine-exposed dams had significantly higher (p<0.001)
levels of apoptosis in the pancreatic islet cells compared to
the offspring of the saline-exposed dams (Table 1). To
confirm that the apoptotic islet cells were in fact insulin-
secreting cells, pancreatic tissue from animals at PND1 was
dual-labelled with insulin and TUNEL staining. In the
pancreas tissue from control offspring there were 8.8±
1.21% TUNEL/insulin-positive beta cells and in the nic-
otine-exposed offspring there were 27.8±3.1% TUNEL/
insulin-positive beta cells (p<0.001).
Discussion
Recent epidemiological studies have shown that the
offspring of women who smoke during pregnancy have
an increased risk of developing obesity, hypertension and
type 2 diabetes [18]. The mechanism(s) underlying these
associations are unknown. The results from this study
demonstrate for the first time that nicotine exposure alone,
at concentrations which are representative of human
exposure [17, 18], during pregnancy and lactation results
in endocrine and metabolic changes in the offspring that are
consistent with the disturbed glucose metabolism that may
lead to type 2 diabetes.
In the present study we have shown that fetal and
neonatal exposure to nicotine results in impaired glucose
homeostasis in response to a glucose challenge prior to a
measurable difference in body weight (7 weeks of age),
suggesting that in this model the metabolic changes asso-
ciated with the onset of type 2 diabetes precede obesity.
Furthermore, we found that nicotine exposure during the
fetal period alone resulted in a significant reduction in
serum insulin concentrations at PND1 in association with a
significant increase in the number of apoptotic islet cells.
We therefore propose that the nicotine-induced damage to
the beta cell during fetal development results in permanent
changes in pancreatic function, which are evident as
impaired glycaemic control at 7 weeks of age and which
persist until adulthood (26 weeks of age).
Similarly, other in utero insults including glucocorticoid
administration, uteroplacental insufficiency and fetal
undernutrition have been shown to result in long-lasting
functional deficiencies in the endocrine pancreas [1922].
At 26 weeks of age nicotine-exposed offspring exhibit
dysglycaemia and increased serum insulin concentrations.
The elevated insulin concentrations at 26 weeks of age are
probably a result of the dysglycaemia and/or impaired
sensitivity to insulin, in a similar manner to glucose intol-
erance in humans. Because dysglycaemia precedes obesity
in this model, we speculate that the metabolic changes
associated with type 2 diabetes are not caused by obesity,
but that this initial underlying beta cell defect could
represent a common antecedent for the postnatal obesity
and the dysglycaemia observed by us.
We propose that nicotine acts directly at the fetal beta
cell to cause apoptosis, possibly by increasing oxidative
stress at that location. In adult rats, nicotine exposure has
been shown to produce oxidative tissue injuries in vivo [23,
24] and can induce oxidative stress in pancreatic tissue in
vitro [25]. Oxidative stress has been shown to decrease
insulin secretion and to increase apoptosis in beta cells
[reviewed in Ref. 26], results which are reflected by the
decrease in serum insulin concentrations and increased beta
cell apoptosis observed at PND1 in this study. Alternatively
nicotine may act directly at the nicotinic receptors on the
beta cell to inhibit insulin secretion, as has been demon-
strated in isolated rat islets in vitro [27]. Further studies will
be required to address this question and to determine the
specific developmental stages that are sensitive to nicotine
exposure.
Results from this study have shown that (1) nicotine, at
levels that are relevant for human exposure, results in
postnatal obesity and impaired glucose homeostasis and (2)
that these postnatal changes may be a result of an under-
lying defect in the pancreatic beta cell. On the basis of our
results, we propose that the long-term postnatal health
consequences of nicotine exposure warrant further inves-
tigation. Our results also raise concerns regarding the
safety of nicotine replacement therapy during pregnancy
and lactation.
Acknowledgements This work was supported by an operating
grant from the Canadian Institutes of Health Research in Nutrition,
Metabolism and Diabetes to A. C. Holloway and K. M. Morrison
(MOP 69025), and a fellowship from the Heart and Stroke
Foundation to K. M. Morrison. H. C. Gerstein holds the Population
Health Institute Chair in Diabetes Research (sponsored by Aventis).
We thank Ms Sandra Stals, Ms Lisa Kellenberger and the staff of the
McMaster University Central Animal Facility for their excellent
technical assistance.
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    • "The placenta is particularly susceptible to ER stress due to its high protein secretory activity, and augmented ER stress has been demonstrated to be associated with adverse placental development and fetal growth restriction (Yung et al. 2012, Kawakami et al. 2014, Yang et al. 2015). Interestingly, similar placental and fetal outcomes were reported in animal models of nicotine exposure during pregnancy (Holloway et al. 2005, Holloway et al. 2014, Gruslin et al. 2009 ). Indeed, nicotine administration during pregnancy increased placental ER stress Nicotine and reproduction R189 resulting in the activation of the UPR at embryonic day 15 (Wong et al. 2015). "
    [Show abstract] [Hide abstract] ABSTRACT: Nicotine exposure during pregnancy through cigarette smoking, nicotine replacement therapies or e-cigarette use continues to be a widespread public health problem, impacting both fetal and postnatal health. Yet, at this time, there remains limited data regarding the safety and efficacy in using these nicotine products during pregnancy. Notably, reports assessing the effect of nicotine exposure on postnatal health outcomes in humans, including reproductive health, are severely lacking. Our current understanding regarding the consequences of nicotine exposure during pregnancy is limited to a few animal studies, which do not comprehensively address the underlying cellular mechanisms involved. This paper aims to critically review the current knowledge from human and animal studies regarding the direct and indirect effects (e.g. obesity) of maternal nicotine exposure, regardless of its source, on reproductive outcomes in pregnancy and postnatal life. Furthermore, this review highlights several key cellular mechanisms involved in these adverse reproductive deficits including oxidative stress, inflammation, and endoplasmic reticulum (ER) stress. By understanding the interplay of the cellular mechanisms involved, further strategies could be developed to prevent the reproductive abnormalities resulting from exposure to nicotine in utero and influence informed clinical guidelines for pregnant women.
    Full-text · Article · Oct 2015 · Reproduction
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    • "Some studies associate metabolic illness, such as obesity and their comorbidities, with an increase in oxidative stress (Bullon et al. 2000; Tariq et al. 2014). However, there are few reports showing the close association between neonatal oxidative stress and the development of diseases at adulthood (Holloway et al. 2005; Bruin et al. 2008; Niu et al. 2013). Habbout and colleagues (2012 and 2013a) showed that increased plasma and cardiac oxidative stress were associated with higher susceptibility to cardiac injury at adulthood . "
    [Show abstract] [Hide abstract] ABSTRACT: Neonatal overfeeding induced by litter size reduction leads to further obesity and other metabolic disorders, such as liver oxidative stress and microesteatosis at adulthood. We hypothesized that overfeeding cause an early redox unbalance at weaning, which could program the animals to future liver dysfunction. Thus, we studied lipogenesis, adipogenesis, catecholamine status and oxidative balance in weaned overfed pups. To induce early overfeeding, litters were adjusted to 3 pups at the 3(rd) day of lactation (SL group). The control group remained with 10 pups per litter until weaning (NL group). The peripheral autonomic nerve function was determined in vivo at 21 days-old. Thereafter, pups were killed for further analysis. Significant differences had P < 0.05. The SL pups presented higher visceral adipocyte area, higher content of lipogenic enzymes (ACC, FAS) with lower content of adipogenic factors (CEBP, PPARγ) in visceral adipose tissue (VAT). Although the autonomic nerve activity and adrenal catecholamines production were not significantly altered, catecholamine receptor (β3ADR) was lower in VAT. They presented higher triglyceride, PPARγ, PPARα and PGC1α contents in liver. In plasma and liver, the SL pups showed an oxidative unbalance, with higher lipid peroxidation and protein oxidation. The SL group presented higher serum ALT. The early increased lipogenesis at adipose tissue and liver in weaned overfed rats, suggest that the higher oxidative stress and lower catecholamine effect in VAT is associated with the early development of liver dysfunctions and adipocyte hypertrophy. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.
    Full-text · Article · Sep 2015 · The Journal of Physiology
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    • "In the present study, we observed an increase of body mass and visceral fat mass in adult S animals. Our data corroborate the literature, which demonstrated the association between neonatal exposure to cigarette smoke or nicotine with obesity development in adulthood (von Kries et al., 2002; Wideroe et al., 2003; Holloway et al., 2005; Goldani et al., 2007; Somm et al., 2008 Somm et al., , 2009 Oliveira et al., 2009; Koshy et al., 2010; Durmus et al., 2011; Santos-Silva et al., 2013). Silva et al. (2013) showed glucose intolerance , characterized by higher fasting glucose with normoinsulinemia in animals programed by cigarette smoke with a higher dose of nicotine than in the present study. "
    [Show abstract] [Hide abstract] ABSTRACT: Children from pregnant smokers are more susceptible to become obese adults and to become drug or food addicts. Drugs and food activate the mesolimbic reward pathway, causing a sense of pleasure that induces further consumption. Here, we studied the relationship between tobacco smoke exposure during lactation with feeding, behavior and brain dopaminergic reward system parameters at adulthood. Nursing Wistar rats and their pups were divided into two groups: tobacco smoke-exposed (S: 4 times/day, from the 3(rd) to the 21(th) day of lactation), and ambient air-exposed (C). On PN175, both offspring groups were subdivided for a food challenge: S and C that received standard chow (SC) or that chose between high-fat (HFD) and high-sucrose diets (HSD). Food intake was recorded after 30 min and 12 h. Offspring were tested in the elevated plus maze and open field on PN178-179; they were euthanized for dopaminergic analysis on PN180. SSD (self-selected diet) animals presented a higher food intake compared to SC ones. S-SSD animals ate more than C-SSD ones at 30 min and 12 h. Both groups preferred the HFD. However, S-SSD animals consumed relatively more HFD than C-SSD at 30 min. No behavioral differences were observed between groups. S animals presented lower tyrosine hydroxylase (TH) content in the ventral tegmental area, lower TH, dopaminergic receptor 2, higher dopaminergic receptor 1 contents in the nucleus accumbens and lower OBRb in hypothalamic arcuate nucleus. Tobacco-smoke exposure during lactation increases preference for fat in the adult progeny possibly due to alterations in the dopaminergic system. Copyright © 2015. Published by Elsevier Ltd.
    Full-text · Article · Jun 2015 · Neuroscience
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