Placenta 35 (2014) 411e416

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Placenta
journal homepage: www.elsevier.com/locate/placenta

Oxidative stress and maternal obesity: Feto-placental unit interaction

N. Malti a, H. Merzouk a, *, S.A. Merzouk b, B. Loukidi a, N. Karaouzene a, A. Malti c, M. Narce d
a
Laboratory of Physiology, Physiopathology and Biochemistry of Nutrition, Department of Biology, Faculty of Natural and Life Sciences, Earth and Universe,
University ABOU-BEKR BELKAD, Tlemcen 13000, Algeria
b
Department of Technical Sciences, Faculty of Engineering, University ABOU-BEKR BELKAD, Tlemcen 13000, Algeria
c
Gynecology and Obstetrics Department, Mother e Infant Hospital Center, University of Tlemcen, 13000, Algeria
d
INSERM UMR 866, Lipids Nutrition Cancer, University of Burgundy, Faculty of Life, Earth and Environment Sciences, Dijon 21000, France

a r t i c l e i n f o

a b s t r a c t

Article history:
Accepted 10 March 2014

Objective: To determine oxidative stress markers in maternal obesity during pregnancy and to evaluate
feto-placental unit interaction, especially predictors of fetal metabolic alterations.
Patients and methods: 40 obese pregnant women (prepregnancy BMI > 30 kg/m2) were compared to 50
control pregnant women. Maternal, cord blood and placenta samples were collected at delivery.
Biochemical parameters (total cholesterol and triglycerides) and oxidative stress markers (malondialdehyde, carbonyl proteins, superoxide anion expressed as reduced Nitroblue Tetrazolium, nitric oxide
expressed as nitrite, reduced glutathione, catalase, superoxide dismutase) were assayed by biochemical
methods.
Results: Maternal, fetal and placental triglyceride levels were increased in obese group compared to
control. Maternal malondialdehyde, carbonyl proteins, nitric oxide and superoxide anion levels were
high while reduced glutathione concentrations and superoxide dismutase activity were low in obesity. In
the placenta and in newborns of these obese mothers, variations of redox balance were also observed
indicating high oxidative stress. Maternal and placental interaction constituted a strong predictor of fetal
redox variations in obese pregnancies.
Discussion: Maternal obesity compromised placental metabolism and antioxidant status which strongly
impacted fetal redox balance. Oxidative stress may be one of the key downstream mediators that initiate
programming of the offspring.
Conclusion: Maternal obesity is associated with metabolic alterations and dysregulation of redox balance
in the mother-placenta e fetus unit. These perturbations could lead to maternal and fetal complications
and should be carefully considered.
2014 Elsevier Ltd. All rights reserved.

Keywords:
Pregnancy
Obesity
Mother
Newborn
Placenta
Oxidative stress

1. Introduction
Obesity is a public health problem in most industrialized
countries, and is now expanding to developing countries [1]. The
rise in the prevalence of obesity might result from the increasingly
sedentary lifestyle associated with a reduction in daily physical
activity and/or from changes in eating behavior, both quantitatively
and qualitatively that may play a role in early childhood [2]. Obesity
is a risk factor for the development of several chronic diseases such
as cardiovascular and respiratory diseases, diabetes type II, hypertension and some forms of cancer [3]. The importance of the intrauterine and neonatal metabolic environment as possible

* Corresponding author. Tel.: 213 778303645.

E-mail address: hadamerzouk_2@hotmail.com (H. Merzouk).
http://dx.doi.org/10.1016/j.placenta.2014.03.010
0143-4004/ 2014 Elsevier Ltd. All rights reserved.

teratogenic determinants for the predisposition of obesity is widely

supported [4]. Furthermore, obesity during pregnancy is associated
to several maternal and fetal complications [5e7]. In addition,
maternal obesity is important in promoting obesity in offspring,
reecting epigenetic programming [8,9]. Placental metabolic
functions are not immune to this imprint [10,11]. Placental
dysfunction is implicated in most of the poor pregnancy outcomes
associated with maternal obesity, and is also known to be involved
in developmental programming of later-life diseases [12,13].
Several studies have reported that obesity is associated with
oxidative stress related to inadequate antioxidant defenses and
increased rates of free radical formation [14]. The impact of
oxidative stress on the fetus and the newborn is well known [15].
Pregnancy is a state of oxidative stress [16]. Placenta tissue is rich in
hormones and is an important source of pro-oxidant agents as well
as antioxidant enzymes [17]. Moreover, oxidative stress may affect

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N. Malti et al. / Placenta 35 (2014) 411e416

the embryonic development [18]. Women with preeclampsia have

an increased oxidative stress and lipid peroxidation and at the same
time have a deciency in several important antioxidants [19]. In
this pathologic pregnancy, a heightened level of oxidative stress is
encountered leading to placental oxidative damage which can
affect placental function [20]. Oxidative stress may lead to a change
in insulin sensitivity, oxidation of lipoproteins, lipid alterations and
adult diseases, such as diabetes and atherosclerosis in the offspring
of obese mothers. Maternal and fetal oxidative stress was enhanced
during obesity [21]. Nevertheless, the simultaneous existence of a
placental oxidative stress and maternal-feto-placental interactions
during maternal obesity are still not well understood.
The aim of the present work was to determine oxidative stress
markers during pregnancy associated with maternal obesity, and to
assess the impact on the maternal-placental-fetal unit. Therefore, it
would be possible to characterize maternal and placental oxidant/
antioxidant imbalance and its impact on fetal redox status in
obesity.
2. Patients and methods
2.1. Study population
The study population included 50 normal weight (prepregnancy BMI between
19 and 25 kg/m2) and 40 obese (prepregnancy BMI  30 kg/m2) women who gave
birth at the Obstetrics and Gynecology department of Tlemcen Hospital, Tlemcen
city (West Algeria). The rigorous selection, recruitment and monitoring of pregnant
women was done by clinicians of the service. These women are aged between 20
and 35 years and have full-term pregnancies (37 weeks). None of the subjects
selected in this study had history of chronic diseases, hypertension, eclampsia, infections or fetal anomalies. All women were routinely seen by an obstetrician and an
endocrinologist. They were tested for gestational diabetes according to the World
Health Organization criteria and all had normal glucose tolerance test during the
third trimester and within 48 h of delivery. All women had uncomplicated singleton
pregnancies and delivered by caesarean section. The indications for elective
Caesarean section at term were breech presentation, placenta praevia and previous
Caesarean section. Maternal and neonatal characteristics are shown in Table 1.
The study was performed according to the Declaration of Helsinki. All participants in this study were informed about the goals and the work in progress, and
were asked to give their written consent beforehand. Investigations of patients as
well as blood and placenta sampling conditions were subjected to a strict code of
ethics. The protocol was approved by the Tlemcen Hospital Committee for Research
on Human Subjects.
2.2. Blood samples
Fasting maternal blood samples were obtained from the arm veins of the
mothers, at the time of delivery. Cord blood samples were obtained from the umbilical vein immediately following delivery and after the cutting of the umbilical
cord. Blood samples were collected in EDTA tubes, were centrifuged and plasma was
separated for assessing lipids and plasma oxidative stress markers. The remaining

Table 1
Maternal and neonate characteristics.
Characteristics

Control

Obese

Number
Age (years)
Prepregnancy BMI (kg/m2)
Number of gestations
Parity
Gestational age (weeks)
Birth weight (Kg)
M/F sex ratio
Total cholesterol
Mothers (g/L)
Newborns (g/L)
Placenta (mg/g)
Triglycerides
Mothers (g/L)
Newborns (g/L)
Placenta (mg/g)

50
29  5.25
22.61  2.13
2.81  1.33
2.69  1.30
38  1
3.34  0.31
28/22

40
31.31  5.91
33.17  3.40**
3.56  2.00
3.12  1.63
38  1
4.28  0.35*
23/17

1.88  0.35
0.55  0.06
5.18  0.64

1.79  0.21
0.46  0.05
5.73  0.53

1.43  0.20
0.41  0.06
6.59  0.51

2.28  0.11**
0.68  0.07*
9.94  0.47*

Values are means  SD. BMI: body mass index (weight/height2); M/F: males/females. Signicant differences between obese and control groups are indicated as:
*P < 0.05; **P < 0.01.

erythrocytes were washed and hemolysed by the addition of cold distilled water (1/
4). The hemolysates were appraised for erythrocyte oxidant/antioxidant status.
2.3. Placental samples
Placentas were collected immediately following delivery, within 10 min of delivery, and washed with saline water to eliminate the blood. Each placenta was
weighed and sampled in three main regions (central region, mid placenta and peripheral region). Placental samples were snap frozen and stored at 80  C until
further analysis.
All samples were homogenized in four volumes of phosphate buffered saline
(PBS) containing proteolytic enzyme inhibitors (Complete-Mini; Roche), using an
Ultra-Turex homogenizer (Bioblock Scientic, Illkirch, France). Samples were then
centrifuged for 30 min at 4000 rpm and the supernatant was collected for
biochemical analysis.
2.4. Determination of biochemical parameters
Plasma or placental homogenate cholesterol and triglycerides were determined
by enzymatic methods (Kits Sigma Chemical Company, St Louis, MO, USA).
2.5. Determination of markers of the oxidant/antioxidant status
Nitric oxide (NO) was determined by the method of Guevara et al. [22], after
plasma or placental homogenate deproteinizing procedure (using methanol:diethylether; 3:1 mixture v/v). Nitrite and nitrate levels were measured
together; nitrate being previously transformed to nitrite by cadmium reduction.
Nitrite was assayed directly spectrophotometrically at 492 nm, using the colorimetric method of Griess (Griess reagent: 1 g/L sulfanilamide, 25 g/L phosphoric acid,
and 0.1 g/L N-1-naphthylethylenediamine). Calibration curves were made with sodium nitrite in concentrations from 1 to 50 mmol/L. The inter- and intra-assay variations were 3.40% and 2.50%, respectively. The determination of the superoxide
anion (O2) was based on Nitro Blue Tetrazolium (NBT) reduction in monofarmazan
by O2 [23]. The blue formazan was dissolved using 2M potassium hydroxide and
dimethylsulfoxide and its formation was monitored spectrophotometrically at
560 nm using the molar extinction coefcient (1.5  104 M1 $ cm1).
The catalase activity (CAT, EC 1.11.1.6) was measured by spectrophotometric
analysis of the decomposition rate of hydrogen peroxide according to the method of
Aebi [24]. The reaction was initiated by addition of placental homogenate or
hemolysate to the reaction mixture containing phosphate buffer (0.05 M, pH 7.2)
and H2O2. Change in absorbance was recorded spectrophotometrically at 240. The
results were expressed as unit of catalase activity corresponding to mmol of H2O2
decomposed per minute using the H2O2 standard curve. The inter- and intra-assay
variations were 2.40% and 3%, respectively.
The assessment of the enzymatic activity of superoxide dismutase (SOD, EC
1.15.1.1) was based on the ability to inhibit pyrogallol autoxidation, with one unit of
SOD activity the amount that causes 50% inhibition of the oxidation of pyrogallol
[25]. SOD activity was measured every 5 min over 1 h at 405 nm, for 1/20 dilution of
placental homogenate and 1/5 dilution of hemolysate. The inter- and intra-assay
variations were 3% and 3.80%, respectively.
Hemolysate or placental homogenate reduced glutathione (GSH) levels were
assayed by a colorimetric method based on the reduction of 5,5-dithiobis-(2nitrobenzoic) acid (DTNB) by GSH to generate 2-nitro-5-thiobenzoic acid which
has yellow color, according a Sigma Aldrich Kit (Saint Louis, MO, USA). The absorbance at 412 nm was measured, and the GSH concentration was then determined
with the GSH standard curve. The inter- and intra-assay variations were 1% and
0.50%, respectively.
Plasma or placental homogenate carbonyl proteins (marker of protein oxidation)
by the derivatization of protein carbonyl groups with 2,4-dinitrophenylhydrazine
(DNPH) leading to the formation of stable dinitrophenyl (DNP) hydrazone adducts,
which can be detected spectrophotometrically at 375 nm (Sigma Aldrich Kit (Saint
Louis, MO, USA). Oxidized BSA standard was used for the standard curve. The interand intra-assay variations were 0.40% and 0.60%, respectively.
Plasma or placental homogenate malondialdehyde (MDA, marker of lipid peroxidation) was estimated by the method of Draper and Hadley et al. [26] using
thiobarbituric acid (TBA). Absorbance was measured at 532 nm. The results were
expressed as micromoles of MDA, using the molar extinction coefcient of chromophore (1.56  105 M1 cm1). The inter- and intra-assay variations were 3.50%
and 3%, respectively.
2.6. Statistical analysis
The results are presented as means and standard deviations. A priori power
analysis was performed to determine the sample size, using power and sample size
calculator (Statistical solutions, Sigma). We calculated that a sample of 50 normal
pregnant and 40 obese women would give us a 95% power of detecting a 25% difference in measurements. The results were tested for normal distribution using the
ShapiroeWilk test. After variance analysis, Students t-test was used to compare
means between normal weight and obese groups, for different parameters. The
multiple regression analysis was performed with dependent variables (lipid parameters and markers of oxidant/antioxidant status in neonates) and independent

N. Malti et al. / Placenta 35 (2014) 411e416

variables (lipid parameters, and maternal-placental markers of oxidant/antioxidant
status). B(ES) are the correlation coefcients (standard error) of each independent
variable with the dependent variable. R2 is the coefcient of determination; it
provides the percentage of variance expressed by all variables. Relationships were
signicant for p < 0.05. All tests were performed using STATISTICA4.1program
(StatSoft, Tulsa, OK).

3. Results
3.1. Lipids and redox balance
Maternal, fetal and placental cholesterol levels showed no signicant difference in the obese group compared to the normal weight
group. In contrast, a signicant increase in maternal, fetal and
placental triglyceride levels was found in the obese group (Table 1).
In obese mothers, oxidant/antioxidant status alterations were
marked by a signicant increase in plasma concentrations of
malondialdehyde, carbonyl proteins, nitrite (reecting nitric oxide)
and reduced NBT (reecting anion superoxide production), and a
signicant decrease in reduced glutathione levels and in erythrocyte
superoxide dismutase activity compared to control values (Figs. 1
and 2). In their newborns, the levels of malondialdehyde, nitrite,
reduced NBT as well as erythrocyte activities of catalase and superoxide dismutase were signicantly high compared to the control
values (Figs.1 and 2). Furthermore, in the placenta of obese mothers,
a signicant increase was noted in the levels of malondialdehyde,
carbonyl protein, reduced glutathione and also in antioxidant
enzyme activities (superoxide dismutase and catalase) compared to
values obtained with placentas from control mothers (Figs. 1 and 2).
3.2. Predictors of fetal metabolic alterations
In order to investigate the inuence of maternal and placental
metabolic changes on fetal metabolism, a multiple regression

413

analysis was performed with independent variables or predictors

(maternal-placental parameters) and dependent variables (neonate
parameters). Maternal and placental cholesterol and triglyceride
levels were not signicantly correlated to those of the newborn
while the interaction between these two predictors explained,
respectively 38% and 39% of the variation of these parameters in
control group. Regarding markers of oxidant/antioxidant status, no
correlation was found between maternal, placental and fetal parameters. The mother-placenta interaction did not alter these observations (Table 2).
In the obese group, maternal and placental cholesterol levels
were not correlated with those of the newborn. However, the
mother-placenta association inuenced fetal plasma cholesterol
levels and explained 35% of their variation (Table 3). There was a
strong correlation between, on the one hand, maternal and fetal
plasma levels of triglycerides and malondialdehyde (P 0.007 and
P 0.008, respectively), and on the other hand, placental and fetal
triglyceride and malondialdehyde levels (P 0.005). The mothere
placenta interaction increased the power of the model (P 0.001
and P 0.004, respectively) and explained 64% and 47%, respectively, of the variation of these parameters in the newborn.
Maternal Nitrite and reduced NBT levels were signicantly
correlated with those of newborns in obese group (P 0.01). The
interaction mother-placenta could signicantly predict the changes
in fetal Nitrite and reduced NBT levels (42% and 43%, respectively).
Regarding the antioxidant enzyme activities, though the maternal
values were not associated with those of newborns, the activities of
placental catalase and superoxide dismutase were positively
correlated to the activities of these fetal enzymes (P 0.030 and
P 0.039, respectively). The mother-placenta interaction was
highly signicant; it explained the respective changes (44% and
43%) of catalase and superoxide dismutase activities in newborns of
obese mothers (Table 3).

Fig. 1. Antioxidant status in mothers and their newborns and placentas. Each value represents the mean  standard deviation. GSH: reduced glutathione; SOD: superoxide dismutase. The signicance of the differences between obese and control group was determined by the students t-test after analysis of variance:*P < 0.05. **P < 0.01.

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N. Malti et al. / Placenta 35 (2014) 411e416

Fig. 2. Oxidant status in mothers and their newborns and placentas. Each value represents the mean  standard deviation. CP: carbonyl proteins; MDA: malondialdehyde; NO:
nitric oxide; O2: superoxide anion. The signicance of the differences between obese and control group was determined by the students t-test after analysis of variance:*P < 0.05.
**P < 0.01.

4. Discussion
This study revealed changes in maternal, fetal and placental
redox status as well as the maternal-placental-fetal interactions
during obesity. Obesity is commonly associated with metabolic
changes with high cardiovascular risk [3]. These metabolic alterations increase during pregnancy [5,6]. In our study, neither the
obese women nor their neonates had any complications. These
obese women presented elevated plasma triglyceride levels; whereas
total plasma cholesterol levels were similar to those of normal
weight women. Similar anomalies were noted in newborns and in
placentas of the obese group. Our ndings were in agreement with
those of other authors who showed high serum triglyceride levels in
obese women and their newborns compared with those of control
group [20,27]. Increased estrogens levels and insulin resistance
during pregnancy are responsible for this hypertriglyceridemia [28].
High triglyceride levels in newborns of obese mothers were correlated to their birth weights which were increased compared to
controls [27]. We have previously found that normal weight newborns of obese mothers had triglyceride levels similar to those found
in control newborns [21]. It has been shown that placental lipoprotein lipase (LPL) activity is high in obese pregnancies [29]. In the
placenta, LPL hydrolyzes triglycerides from maternal circulation to
release free fatty acids (FFA), which are transferred to the placenta,
and then esteried to triglycerides. Other authors have noted an
accumulation of triglycerides in the placenta of obese women
[30].The multiple regression analysis in the control group showed
that although maternal or placental lipid levels are not independently correlated to those of the fetus, the maternaleplacental

interaction was a substantial predictor of changes in fetal lipids. This

was quite normal because the fetus is dependent on fat intake from
the mother and transferred via the placenta. In the obese group, the
correlations between maternal, placental and fetal triglycerides
were more important, and the impact of the maternaleplacental
interaction was stronger. This supposed the concept that in the
presence of high maternal triglyceride levels, the placental transfer
was more important, leading to fetal hypertriglyceridemia. Previous
studies revealed also a highly signicant correlation between
maternal and fetal triglycerides [27]. It has been shown that high
levels of triglycerides in maternal circulation may create a steep
concentration gradient across the placenta, which accelerates their
transport and deposition in fetal tissues [9].
Our results provided evidence that oxidant/antioxidant status
was altered in the maternal fetoplacental unit during obesity. In
addition, the multivariate analysis clearly revealed that fetal
changes in lipid peroxidation, free radicals and antioxidant enzymes
were directly related to maternal and/or placental changes in
obesity. Oxidative stress is characterized by an imbalance between
pro-oxidants (free radicals, peroxides) and antioxidants (superoxide
dismutase, catalase, glutathione peroxidase, antioxidant vitamins)
[31,32]. Today, it is well established that oxidation phenomena are
involved in the development of metabolic and neurodegenerative
diseases, as well as aging. Vincent et al. . [33] showed that obesity
increases oxidative stress by raising lipid peroxidation proportionally to the degree of adiposity and low antioxidant defense. Significant changes of the oxidant/antioxidant balance still exist during
normal pregnancy, due to increased basal oxygen and energy consumption in different organs including the fetoplacental unit [16].

N. Malti et al. / Placenta 35 (2014) 411e416

Table 2
Multivariate analysis in the control group.
Dependent Variables
(newborns)
Cholesterol
B (ES)
P
R2
Triglycerides
B (ES)
P
R2
Malondialdhyde
B (ES)
P
R2
Carbonyl proteins
B (ES)
P
R2
Nitrite
B(ES)
P
R2
red. NBT
B (ES)
P
R2
Catalase
B (ES)
P
R2
Superoxide dismutase
B (ES)
P
R2
Reduced glutathione
B (ES)
P
R2

Mothers
variables

Placenta
variables

Interaction
Mother-placenta

0.132 (0.061)
0.215
e

0.184 (0.053)
0.119
e

0.416 (0.065)
0.015
0.38

0.136 (0.040)
0.138
e

0.194 (0.020)
0.140
e

0.424 (0.069)
0.014
0.39

0.082 (0.050)
0.102
e

0.102 (0.041)
0.147
e

0.155 (0.077)
0.125
0.07

0.147 (0.032)
0.139
e

0.125 (0.051)
0.108
e

0.199 (0.078)
0.089
0.10

0.187 (0.071)
0.107
e

0.117 (0.064)
0.114
e

0.188 (0.027)
0.095
0.11

0.111 (0.037)
0.175
e

0.157 (0.027)
0.116
e

0.158 (0.063)
0.115
0.09

0.142 (0.070)
0.104
e

0.181 (0.030)
0.111
e

0.182 (0.083)
0.101
0.12

0.201 (0.073)
0.108
e

0.128 (0.076)
0.143
e

0.199 (0.051)
0.168
0.07

0.102 (0.052)
0.222
e

0.152 (0.075)
0.177
e

0.165 (0.077)
0.125
0.06

B (ES) are the correlation coefcients (standard error) of each independent variable
with the dependent variable. R2 is the coefcient of determination and provides the
percentage of variance explained by all variables. Relationships are signicant at
p < 0.05.

Antioxidants are extremely important in maintaining cell function

during normal pregnancy [34]. According to Mueller et al. [17],
during normal pregnancy, the placenta is a major source of prooxidants and antioxidant systems; it is capable of keeping lipid
peroxidation under control. The placenta is rich in mitochondria,
highly vascular and is exposed to high maternal oxygen partial
pressure, therefore resulting in increased production of superoxide.
The nitric oxide (NO) is also locally produced by the placenta from a
substrate, L-arginine, by NO synthases (NOS) [35]. The formation of
these two free radicals is tightly controlled by a highly effective
antioxidant defense system during normal pregnancies. However,
under pathological conditions, the redox balance is disturbed, thus
inducing an oxidative stress. In our study, obese mothers showed
high levels of pro-oxidant markers (superoxide anion as expressed
by reduced NBT, nitric oxide as expressed by nitrite, malondialdehyde, carbonyl proteins) and a reduction in antioxidant defenses
(reduced glutathione, superoxide dismutase). In our study,
conventionally methods were used to estimate superoxide anion
and nitric oxide. The reduction of NBT to insoluble blue formazan
was used as a probe for superoxide generation, although it is not
entirely specic for O2. Nitric oxide production was estimated
from determining the concentrations of nitrite end products. A
reduction in SOD, the primary enzyme that inactivates the superoxide radical and in GSH which is involved in glutathionedependent enzyme reactions, would lead to increased numbers of
free radicals and this could thereafter be responsible for the

415

increased levels of malondialdehyde and carbonyl proteins in obese

mothers. Antioxidant enzymes may also be consumed or inactivated
in high oxidative conditions. Several studies reported that oxidative
stress status in obese women during pregnancy was always associated with a pro-inammatory state [36]. Our results proved the
presence of oxidative stress in placentas of obese mothers. It is
important to note that measurements were taken on term placentas
at a single time point. The oxidative stress was characterized by high
levels of malondialdehyde and carbonyl proteins despite elevated
levels of reduced glutathione. Placental levels of nitric oxide (nitrite)
and superoxide anion (reduced NBT) did not change signicantly
between the two groups. This might have resulted from the parallel
increase in the activities of antioxidant enzymes, catalase and superoxide dismutase. High placental antioxidant activities might be a
compensatory mechanism that limits oxidative stress in the
placenta, during obesity [37]. Maintaining normal placental nitric
oxide concentrations is an important aspect for a good pregnancy
development, as nitric oxide plays a signicant role in placental
development, angiogenesis and vasodilatation [35]. However, in the
placenta, excess superoxide and nitric oxide production can result in
peroxynitrite formation, leading to nitrative stress. Peroxynitrite is a
powerful pro-oxidant capable of initiating lipid peroxidation and
nitro-tyrosine formation [19,37]. Further studies are then needed to
conrm this hypothesis in the placenta of obese mothers.
In the present study, infants of obese mothers were also subjected to oxidative stress. There was a signicant increase in
malondialdehyde, superoxide anion and nitric oxide levels in
newborns of obese mothers compared to those of normal-weight
mothers. There is some evidence that maternal obesity induced
mitochondrial dysfunction with an increase in mitochondrial
reactive oxygen species and oxidative stress in oocytes, zygotes and
embryonic life [9]. Our ndings are in agreement with the hypothesis that mitochondrial injury due to maternal obesity could
compromise metabolism in the developing fetus and may even
impact fetal mitochondrial function prior to conception.
In newborns of obese mothers, the increase in pro-oxidants was
associated with a parallel increase in antioxidant enzymes activities,
catalase and superoxide dismutase. The over expression of antioxidant activities in these infants might be an adaptive response with
an induction to counter the effect of increased oxidative stress. Our
results suggested that these infants were exposed to greater
oxidative stress despite higher antioxidant enzyme activities.
The multivariate analysis showed that, during normal pregnancy, fetal redox status was not correlated with that of the mother
or with that of the placenta, suggestive of a maximum protection of
the fetus against free radicals. Pro-oxidants from the mother or
from the placenta itself are usually destroyed before they reach
child development [38]. In contrast, during obesity, the multivariate analysis indicated that the correlations between maternal or
placental redox status and that of the fetus were signicant. The
maternaleplacental interaction was an important predictor of
changes in fetal oxidant/antioxidant status during obese pregnancies. It was clear that oxidative stress in obese mother induced
placental and fetal oxidative stress despite high placental antioxidant defense. Oxidative stress associated with obesity during
pregnancy may be a contributing factor in postnatal consequences
of the neonate and the induction of programmed phenotypes in the
adult offspring. In fact, oxidative stress has also been identied as a
contributing factor in epigenetic mechanisms [7e9,18,19,37,38].
However, only term pregnancies and placentas were considered in
this study. We are then limited in our ability to identify redox alterations during the different trimesters of pregnancy. Further
studies are needed to determine time course of change in maternal
and placental oxidant/antioxidant status during the association of
obesity and pregnancy.

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N. Malti et al. / Placenta 35 (2014) 411e416

Table 3
Multivariate analysis in the Obese group.
Dependent variables
(newborn)
Cholesterol
B (ES)
P
R2
Triglycerides
B (ES)
P
R2
Malondialdehyde
B (ES)
P
R2
Carbonyl proteins
B (ES)
P
R2
Nitrite
B (ES)
P
R2
red. NBT
B (ES)
P
R2
Catalase
B (ES)
P
R2
Superoxide dismutase
B (ES)
P
R2
Reduced glutathione
B (ES)
P
R2

Mother
variables

Placenta
variables

Interaction
mother e placenta

0.111 (0.039)
0.108
e

0.124 (0.026)
0.192
e

0.413 (0.081)
0.019
0.35

0.483 (0.053)
0.007
e

0.547 (0.095)
0.005
e

0.829 (0.028)
0.001
0.64

0.443 (0.065)
0.008
e

0.487 (0.055)
0.005
e

0.526 (0.028)
0.004
0.47

0.142 (0.045)
0.112
e

0.125 (0.054)
0.115
e

0.146 (0.038)
0.107
0.05

0.358 (0.067)
0.010
e

0.136 (0.071)
0.123
e

0.458 (0.044)
0.008
0.42

0.339 (0.046)
0.012
e

0.190 (0.045)
0.118
e

0.480 (0.031)
0.007
0.43

0.172 (0.060)
0.226
e

0.303 (0.082)
0.030
e

0.496 (0.055)
0.005
0.44

0.284 (0.052)
0.070
e

0.328 (0.040)
0.039
e

0.485 (0.078)
0.006
0.43

0.184 (0.078)
0.140
e

0.166 (0.078)
0.116
e

0.209 (0.037)
0.155
0.06

B (ES) are the correlation coefcients (standard error) of each independent variable
with the dependent variable. R2 is the coefcient of determination and provides the
percentage of variance explained by all variables. Relationships are signicant at
p < 0.05.

In conclusion, several metabolic and redox status changes

appeared in the mother, the fetus and the placenta during obesity.
Disturbances in the oxidant/antioxidant status that affected the
entire unit mother-placenta-fetus may be responsible, during
pregnancy, of a range of maternal and fetal complications and may
be one of the key downstream mediators that initiate programming
of the offspring. Reducing maternal oxidative stress will be
important for developing therapeutic strategies for alleviating
long-term programmed consequences associated with obesity.
Acknowledgments
The present work was realized with the nancial support of the
National Agency for the Development of Health Research (PNR
ANDRS). Our thanks go to all volunteers.
None of the authors has any nancial or personal conicts of
interest.
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