Physiology & Behavior 209 (2019) 112587

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Physiology & Behavior

journal homepage: www.elsevier.com/locate/physbeh

Sex differences in glucocorticoids-induced anabolic effects in rats T

a,1 a,1 a
Larissa Rugila S. Stopa , Camila F. de Souza , Geisielle Fernandes Santos ,
Andressa B. Martinsa, Renan Nascimento Ferreirad, Fábio Goulart de Andraded,
Cristiane Mota Leiteb, Dimas A.M. Zaiac, Cassia Thaïs B.V. Zaiaa, Ernane Torres Uchoaa,
⁎

a
Department of Physiological Sciences, State University of Londrina, Londrina, PR, Brazil
b
University of Northern Parana (UNOPAR), Londrina, PR, Brazil
c
Department of Chemistry, State University of Londrina, Londrina, PR, Brazil
d
Department of Histology, State University of Londrina, Londrina, Brazil

GRAPHICAL ABSTRACT

ARTICLE INFO ABSTRACT

Keywords: Glucocorticoids (GC) increase food intake and body weight in humans and rodents and chronic stress and GC
Dexamethasone treatment-induced enhancement of the plasma concentration of GC lead to obesity and metabolic changes. In
Corticosterone response to hypercaloric treatment, males were shown to be more susceptible to obesity than females, de-
Male monstrating that sex differences may affect energy homeostasis. The objective of the current study was to
Female
evaluate the effects of prolonged (28 days) treatment with dexamethasone or corticosterone on food intake and
Metabolism
Food intake
body weight gain in intact rats, both male and female. Also examined were Lee index, weights and area of
adipocytes of retroperitoneal and perigonadal+perirenal adipose tissues, glucose tolerance test (GTT) and
plasma concentrations of free fatty acids, cholesterol and triglycerides. Treatment with dexamethasone was able
to increase body weight, food intake, area of adipocytes and weight of retroperitoneal adipose tissue in males.
Prolonged treatment with corticosterone also stimulated body weight gain and food intake in males. In addition,
it induced an increase in the area of adipocytes and weight of perirenal+perigonadal adipose tissue and higher
glycemia after GTT in these animals, without changes on Lee index and plasma parameters after both GC
treatments. No parameter was changed by dexamethasone or corticosterone treatment in female rats. Thus, it
can be concluded that male rats are more susceptible to the anabolic effects of glucocorticoids than female rats,

Corresponding author at: Department of Physiological Sciences, UEL, State University of Londrina, Rodovia Celso Garcia Cid, PR 445, km 380, Campus
⁎

Universitário, Londrina 860570-970, PR, Brazil.

E-mail address: euchoa@uel.br (E.T. Uchoa).
1
Larissa Rugila S. Stopa and Camila F. de Souza contributed equally to this work.

https://doi.org/10.1016/j.physbeh.2019.112587
Received 21 April 2019; Received in revised form 17 June 2019
Available online 24 June 2019
0031-9384/ © 2019 Elsevier Inc. All rights reserved.
L.R.S. Stopa, et al. Physiology & Behavior 209 (2019) 112587

and these responses can be due to the protective effects of circulating estrogens in females, and/or the difference
between males and females in the expression/activity of corticosteroids receptors.

1. Introduction
Energy homeostasis is regulated by the central nervous system,
especially by the hypothalamus. Peripheral factors also act as reg-
ulators, for example leptin and insulin (often related to adiposity) and
glucocorticoids (GC), the ultimate hormones produced by the hy-
pothalamic-pituitary-adrenal axis [1]. GCs have different functions and
actions and they are particularly important in physiological regulation
and adaptation to stress situations. In cases of chronic stress, it is known
that there is an increase in the plasma concentration of this hormone
that induces increased appetite and body weight gain in humans and
rodents [2]. Increased plasma GC concentration also occurs when
synthetic glucocorticoid is administered as a pharmacological treat-
ment, for example, using dexamethasone.
GCs are steroid hormones, and, in the cytoplasm, they bind to
specific protein receptors—corticosteroid receptors—which are re-
ferred to as mineralocorticoid (MR) or type I receptors and gluco-
corticoid or type II (GR) receptors. It is known that endogenous corti-
costeroids, such as corticosterone in rodents, can bind to both receptors,
while the synthetic glucocorticoid dexamethasone binds almost ex-
clusively to GRs [3].
It is well established in the literature that males have less total white
adipose tissue than females but it is more centrally distributed, con-
tributing to a greater risk of metabolic disorders and their comorbidities
[4]. Females have more total white adipose tissue, but it is more dis-
tributed in the subcutaneous portion. White adipose tissue is more ac-
cumulated in the intra-abdominal depot in men and postmenopausal
women [4]. In view of these facts, it has been shown that, in response to
hypercaloric treatment, male mice were more susceptible to obesity
than intact females. But ovariectomy (bilateral removal of the ovaries)
in females made them as likely to develop obesity as males [5]. This
shows that sex differences may affect energy homeostasis, but studies
are needed to determine the specifics, because the data are currently
sparse and controversial.
In the literature there are no reports on the sex differences on food
intake and energy metabolism induced by either synthetic or en-
dogenous glucocorticoids treatments. Consequently, it is relevant to
study the differential effects of prolonged treatment with glucocorti-
coids on energy homeostasis in male and female animals. The objective
of this study was to evaluate and compare the effects of a 28-day
treatment with dexamethasone or corticosterone on the energy balance
in male and female Wistar rats.
2. Material and methods
2.1. Animals
Male and female Wistar rats (60 days, 220–250 g body weight) were
obtained from the Central Animal Care Facility of the State University
of Londrina. Animals were kept in cages at controlled temperature
(22 ± 2 °C), with a fixed light–dark cycle (light from 6:00 AM to
6:00 PM) and with ad libitum access to pelleted rat chow and fluid,
unless otherwise specified. They were daily monitored for 28 days, for
evaluation of food intake, body weight, and estrus cycles for females.
The work was conducted at the Department of Physiological Sciences
(CIF) at the Center of Biological Sciences, State University of Londrina,
and all experiments were approved by the Committee of Ethics of
Experimental Animals of the UEL (Protocol Number 14638.2016.42).
Animals were organized according to treatment and constituted the
following experimental groups: 1) Females receiving tap water (Female-
Water); 2) Females receiving dexamethasone (DEXA: 0.5 μg/l) diluted
in tap water (Female-DEXA); 3) Males receiving tap water (Male-

Fig. 1. Body weight gain (A, B), and Food intake (C, D) of females and males, which were treated with: Water or dexamethasone (DEXA: 0.5 μg/l, n = 6–12) (A, C)
and Water/ETOH or corticosterone (CORT: 15 mg/l, n = 12–18) (B, D) for 28 days. Data are expressed as mean ± SEM. * P < 0.05.

2
L.R.S. Stopa, et al.
Table 1
Fluid intake and Lee index of female and male animals treated with Water or dexamethasone (DEXA) and Water/ETOH or Corticosterone (CORT).
Female
Water
Fluid intake (ml/100 g) 11.4 ± 0.5
Lee Index (g1/3/cm) 0.308 ± 0.002
Water/ETOH
Fluid intake (ml/100 g) 11.6 ± 0.3
Lee Index (g1/3/cm) 0.288 ± 0.004
Data are expressed as mean ± SEM. N = 12–22.
Water); 4) Males receiving DEXA diluted in tap water (0.5 μg/l) (Male-
DEXA); 5) Females receiving tap water containing 0.5% ethanol
(Female-Water/ETOH); 6) Females receiving tap water containing
corticosterone (CORT: 15 mg/l) diluted in 0.5% ethanol (Female-
CORT/ETOH); 7) Males receiving tap water containing 0.5% ethanol
(Male-Water/ETOH); 8) Males receiving tap water containing CORT
(15 mg/l) diluted in 0.5% ethanol (Male-CORT/ETOH).
2.2. Drugs
Dexamethasone (Ache, Sao Paulo-Brazil) was used in the prolonged
treatment of synthetic glucocorticoid (28 days), being diluted in water
and administered orally at the concentration of 0.5 μg/l, as the only
liquid offered to animals [6]. Corticosterone (Sigma, Saint Louis-USA)
was used in the prolonged treatment of endogenous glucocorticoid
(28 days) and had as dilution vehicle 0.5% ethanol and was adminis-
tered orally at a concentration of 15 mg/l [6]. The solution of corti-
costerone with ethanol was diluted in tap water, which was the only
liquid offered to the animals.
2.3. Experimental design and animal model
Effects of prolonged treatment with dexamethasone and corticos-
terone on food intake, body weight, glucose tolerance test (GTT), Lee
index, plasma metabolic parameters and white adipose tissue in female
and male rats.
Female and male animals received tap water or DEXA, and tap
water/ETOH or CORT, as described above, for 28 days. During this
experimental period, body weight, food and fluid intakes were daily
measured. On the 27th day, animals had their food removed at
8:00 AM, and at 2:00 PM, a drop of blood from the tail was collected for
the determination of basal glycemia by using the test tape of the Accu-
Check Advantage II (Roche, Taquara, RJ, Brazil) device for the de-
termination of blood glucose. Then, intraperitoneal injection of glucose
(1.0 g/kg of body weight) at the concentration 5% was performed, and
blood glucose measurements were done by the test strip 15, 30, 60 and
120 min after glucose overload [6–8]. Food was returned to the cages in
the end of GTT.
On day 28th, animals were also submitted to food restriction at
8:00 AM and Lee index was calculated by considering body weight and
naso-anal distance, whereas the cubic root of body weight in gram was
divided by naso-anal length in centimeters [9]. Rats were euthanized by
decapitation at 2:00 PM, trunk blood was collected in heparinized tubes
and centrifuged at 14,000 ×g for 20 min at 4 °C to obtain the plasma,
which was stored at −20 °C for biochemical analysis. Perigonadal,
perirenal and retroperitoneal adipose tissues were carefully removed
and weighed at autopsy.
2.4. Determination of the estrous cycle
The estrous cycle throughout the experiment was daily evaluated.
Physiology & Behavior 209 (2019) 112587
Male
DEXA Water DEXA
11.7 ± 0.3 12.0 ± 0.2 11.8 ± 0.3
0.307 ± 0.002 0.302 ± 0.002 0.298 ± 0.002
CORT/ETOH Water/ETOH CORT/ETOH
11.7 ± 0.2 11.6 ± 0.3 12.0 ± 0.3
0.291 ± 0.003 0.305 ± 0.003 0.305 ± 0.004
For this, vaginal smear was daily collected at 8:00 AM on a histological
glass slide, which was analyzed in optical microscope at 10× magni-
fication. The phases of the estrous cycle were determined according
with the proportions and types of the cells (cornified epithelial cells,
nucleated epithelial cells and leukocytes). In proestrus, there was pre-
dominance of nucleated epithelial cells, while the estrous phase was
identified by the presence of a greater proportion of cornified epithelial
cells. The metaestrus (diestrus I) was identified by cornified and nu-
cleated cells, accompanied by many leukocytes, and the diestrus
(diestrus II) was characterized by the presence of leukocytes cells. Only
females in estrus or diestrus II on the days of the experiments (GTT or
euthanasia) were considered in this study.
2.5. Histology of visceral white adipose tissue
Perigonadal, perirenal and retroperitoneal subdivisions of visceral
white adipose tissues were carefully removed, weighted immediately
after decapitation, and the values were expressed in g/100 g. After
fixation in 4% formaldehyde in phosphate buffered saline (pH 7.4), for
48 h at 4 °C, adipose tissues were first dehydrated in graded ethanol,
then cleared in xylol, and embedded in paraffin at 62 °C. Using mi-
crotome, sections of 12 mm were obtained, stained by hematoxylin and
eosin, and the images were captured with an image system attached to
the microscope (Motic, Xiamen, China). Using image J, images were
analyzed and the areas of adipocytes (μm2) were determined, as pre-
viously described [6].
2.6. Measurement of cholesterol, triglycerides and free fatty acids plasma
levels
Cholesterol and triglyceride plasma concentrations were de-
termined by spectrophotometry using the BioLiquid Cholesterol
Commercial KIT (Laborclin, PR, Brazil) and Triglycerides BioLiquid KIT
(Laborclin, PR, Brazil), respectively, according to the instructions of the
manufacturers. The results obtained from plasma concentrations of
total cholesterol and triglycerides were expressed in mg/dl. The con-
centration of free fatty acids (FFA) was determined using the spectro-
photometric method of Falholt et al. [10] and the values were expressed
in μmoles/dl. The concentration of corticosterone expressed in μg/dl
was obtained by the fluorometric method of Guillemin et al. [11], based
on the fluorescence of corticosterone in sulfuric acid. The analysis was
performed on a fluorimeter (Victor3TM, PerkinElmer) with excitation
at 477 nm, emission 520 nm and sensitivity 11.
2.7. Statistical analysis
Data were expressed as means ± standard error of the mean (SEM).
The effects of glucocorticoid treatments with their respective controls
(Water or Water/ETOH) were compared in females in diestrus II, fe-
males in estrus and males. Shapiro-Wilk normality test was first per-
formed, and data were analyzed by t-test or Mann-Whitney test, when
3
L.R.S. Stopa, et al. Physiology & Behavior 209 (2019) 112587

Fig. 2. Weights of perirenal + perigonadal (A, B) and retroperitoneal (C, D) adipose tissues of female and male rats treated with: Water or dexamethasone (DEXA:
0.5 μg/l, n = 6–12) (A, C) and Water/ETOH or corticosterone (CORT: 15 mg/l, n = 12–18) (B, D) for 28 days. Data are expressed as mean ± SEM. *P < 0.05.

4
L.R.S. Stopa, et al. Physiology & Behavior 209 (2019) 112587

appropriate. Differences were considered significant when P < 0.05.
3. Results
Effects of prolonged treatment with dexamethasone and corticos-
terone on food and fluid intakes, body weight, glucose tolerance test
(GTT), Lee index, plasma metabolic parameters and white adipose
tissue in female and male rats.
The results obtained show that there was an increase (P < 0.05) in
body weight and food intake in male groups treated with DEXA or
CORT, compared to their respective control group (Fig. 1A). Con-
versely, DEXA or CORT treatment did not change body weight and food
intake in female animals. In DEXA and CORT protocol, there was no
difference in fluid intake among the groups (Table 1). In both sexes
there was no difference in the Lee index between animals given glu-
cocorticoids treatments and their respective control groups (Table 1).
In Figs. 2 and 3 it can be observed that the male group treated with
DEXA presented greater (P < 0.05) relative weight and area of the
adipocytes of the retroperitoneal adipose tissue (g/100 g) when com-
pared to the Male-Water group. DEXA also induced increase in the
weight of perigonadal+perirenal adipose tissue, without difference in
the area of adipocytes of this fat depot. Figs. 2 and 3 also show that
there was an increase (P < 0.05) in the relative weight and area of the
adipocytes of perigenal+perirenal tissue in male rats treated with
CORT, compared to the Male-Water/ETOH group, without statistical
differences in the retroperitoneal adipose tissue. In females, neither

Fig. 3. Representative photomicrographs (100× magnification) of retroperitoneal (A–D) and perirenal + perigonadal (E–H) adipose tissues, stained with hema-
toxylin-eosin, of female and male groups, treated with Water or dexamethasone (DEXA: 0.5 μg/l) (A–D) and Water/ETOH or corticosterone (CORT: 15 mg/l) (E–H).
respectively.

5
L.R.S. Stopa, et al.
Fig. 4. Glucose tolerance test (GTT) (A, B and C) and area under the curve (AUC) of GTT (D) of females (diestrus II or estrus) and males, which were treated with
Water or dexamethasone (DEXA: 0.5 μg/l, n = 6–12) for 28 days. Data are expressed as mean ± SEM. * P < 0.05.
DEXA nor CORT changed the weights and area of the adipocytes of the
two visceral white adipose tissues, compared to their respective control
group animals.
It can be noted that the glycemic profile presented by the animals
treated with DEXA was similar to the profile of the animals treated with
water, both in males and females in diestrus (Fig. 4A, C), while females
in estrus treated with DEXA showed lower glycemia after 60 min of
glucose overload (Fig. 4B). Statistical AUC of GTT analysis showed no
significant difference between treatments in both sexes (Fig. 4D). Re-
garding treatment with CORT, females in diestrus II presented lower
basal glycemia (Fig. 5A). In response to glucose overload, it can be
observed that after the treatment with CORT, females in estrus had a
lower glycemic value after 60 min of glucose injection (Fig. 5B), and
males treated with CORT had higher glycemic values after 60 and
120 min of glucose overload (Fig. 5C). The AUC GTT analysis of the
animals treated with CORT showed that females in estrus treated with
this glucocorticoid had lower glycemia compared to those treated with
water. Males treated with CORT presented higher glycemia than those
treated with water (Fig. 5D).
6
Physiology & Behavior 209 (2019) 112587
As can be seen in the Fig. 6, DEXA or CORT treatments did not
change total cholesterol, triglycerides, free fatty acids and corticos-
terone plasma concentrations, compared to their respective groups
treated with water, both in males and females, in diestrus II or estrus.
4. Discussion
Data from the present study show that treatment of male rats with
dexamethasone, a synthetic glucocorticoid, or corticosterone, an en-
dogenous glucocorticoid, promoted an increase in food intake, body
weight, weight and area of adipocytes of visceral adipose tissue, and
glucose intolerance—but there were none of these effects in females.
However, in both sexes, GCs did not promote changes in the other
parameters evaluated such as Lee index, plasma concentrations of tri-
glycerides, total cholesterol, and free fatty acids.
Though prolonged treatments with DEXA and CORT did not alter
the Lee index, the increase in body weight associated with higher food
intake in male rats, but not in females, may be supported by other
studies which demonstrated the effectiveness of dexamethasone and
L.R.S. Stopa, et al.
Fig. 5. Glucose tolerance test (GTT) (A, B and C) and area under the curve (AUC) of GTT (D) of females (diestrus II or estrus) and males, which were treated with
Water/ETOH or corticosterone (CORT: 15 mg/l, n = 12–18) for 28 days. Data are expressed as mean ± SEM. * P < 0.05.
corticosterone in increasing food intake and body weight gain in male
rodents and humans [2,12,13]. Indeed, previous studies using GC
treatments at other concentrations showed different results regarding
weight gain and food intake. With DEXA, for example, it was shown
that at 2 mg/ml diluted in water, the animals presented lower body
weight and food intake than the control group [14]. In addition, DEXA,
when administered at a dose of 1 mg/kg of body weight, in-
traperitoneally, also caused a decrease in body weight and food intake
[15]. This difference is associated with the different concentrations or
doses of the glucocorticoid. Data from the literature have shown that
the effect of this hormone on food intake is determined by the dose of
administered GC—low doses stimulating food intake, body weight gain
and lipid deposition and high doses reducing dietary intake and pro-
moting lipid and protein catabolism and, consequently, the reduction of
body weight [16]. Accordingly, the concentrations used in the present
study [6] can be considered low and therefore promoted an increase in
body weight and food intake in male animals, but not in intact females.
The increase in the visceral adiposity in males in response to GC
treatment might be ascribed to the stimulation of lipoprotein lipase
enzyme activity by GC [17]. Indeed, visceral adipose tissues are very
susceptible to the anabolic actions of GC due to the high amount and
7
Physiology & Behavior 209 (2019) 112587
affinity of corticosteroid receptors in this fatty tissue deposit [18,19].
Thus, the difference in DEXA and CORT response in different visceral
adipose tissues may be a consequence of the differential predominance
of GR and MR expression in each tissue, since DEXA has more affinity
for GR but CORT can bind to both receptors. However, further studies
are necessary to investigate this hypothesis. Again, there was no effect
of GC on visceral adipose tissue weights evaluated in females.
In fact, it is known that treatment with CORT (100 mg/l) or DEXA
(0.25 mg/kg day) for four weeks in male rodents [13,19,20] resulted in
increased concentrations of glucose, triglycerides, cholesterol, plasma
insulin, and induced glucose intolerance and insulin resistance. Except
for GTT and AUC of the GTT of the animals treated with CORT, the
results of the present study partially diverge from these data in the
literature, as it was observed that animals treated with DEXA and CORT
did not present alteration in most of these parameters analyzed in the
present study. This difference can be attributed to the higher dose/
concentration of GC used in the studies cited. Alternatively, corticos-
terone-induced impairment in glucose tolerance in males may be a re-
sult of the lower GLUT4 translocation to the cell surface, especially in
the skeletal muscle, reducing peripheral uptake of glucose [21–24].
As has been stated in the methodology section and description of the
L.R.S. Stopa, et al. Physiology & Behavior 209 (2019) 112587

(caption on next page)

8
L.R.S. Stopa, et al.
Fig. 6. Plasma levels of total cholesterol (A, B), triglycerides (C, D), free fatty acids (E, F) and corticosterone (G, H) of females (diestrus II or estrus) and males, which
were treated with: Water or dexamethasone (DEXA: 0.5 μg/l, n = 6–12) (A, C, E) and Water/ETOH or corticosterone (CORT: 15 mg/l, n = 12–18) (B, D, F) for
28 days. Data are expressed as mean ± SEM.
results, two phases of the estrous cycle (diestrus II and estrus) were
selected for the evaluation of the effects of GC specifically on the me-
tabolic parameters in plasma. However, the same animals used in the
analysis of the plasma parameters were also used for the other para-
meters but forming only one group (females = diestrus+estrus). Thus,
as pointed out in the above paragraphs, the anabolic effects of GCs
observed in males in the various parameters evaluated in the present
study were not observed in females, neither in diestrus II nor estrus
cycle phase. In this context, a recent study of the group showed that
treatment with CORT and DEXA in the same concentrations as the
current study also demonstrated that these glucocorticoids did not
change these parameters when evaluated in female rats in diestrus I [6].
In addition, CORT treatment (400 μg/ml), for 21 days, reduced body
weight in male and female rats [25], and treatment with a high dose of
dexamethasone (1 mg/kg), for 5 days, decreased both food intake and
body weight gain in rats of both sexes [15]. Despite the divergent data
with those of the present study due to the higher concentration of
corticosterone and dose of dexamethasone administered, the first work
mentioned [25] did not aim to evaluate the energy homeostasis nor the
sex differences within this context. The second one focused on age- and
gender-related changes in glucose homeostasis in glucocorticoid-treated
rats, but it did not use females in specific phases of the estrus cycle. We
cannot rule out that higher doses of glucocorticoids could also induce
metabolic changes not only in male animals but also in females, since
the metabolic effects of glucocorticoids are dependent on the dose/
concentration used in each study.
Thus, the present study demonstrates, for the first time in the lit-
erature, that male rats are more susceptible to the anabolic effects of GC
(endogenous and synthetic) than females. In fact, Hong et al. [5] de-
monstrated that male animals were more likely to develop obesity than
females in response to a hyperlipidic diet. Accordingly, the current
study is reinforced by data in the literature that demonstrated higher
food intake and body weight gain induced by stress in male rats com-
pared to females, strongly suggesting that males are more responsive to
the anabolic effects of stress, which is known to induce marked release
of glucocorticoids [26]. Furthermore, Santos and colleagues [15] ob-
served that a high dose of dexamethasone induced glucose intolerance
in adult (3 months) male rats, but not in adult females—but the same
treatment made older female rats intolerant to glucose. The authors
suggested that the decreased plasma estradiol levels and/or estradiol
receptor responsiveness could account for their increased vulnerability
to modulation of glucose homeostasis by GC. In addition, this sex dif-
ference in the response to anabolic treatments can be explained by the
higher concentration of circulating estrogens in females, since it is
known that estrogens have a protective effect on the development of
obesity, as well as on their comorbidities, induced by a hyperlipidic diet
in males [27], intact females [28] and ovariectomized animals [29,30].
Accordingly, Souza et al. [6] demonstrated that ovariectomy increased
the susceptibility to glucocorticoids-induced anabolic changes, and es-
tradiol treatment could prevent these effects. However, it would not be
wise to exclude the possibility that the increased responsiveness of
males to the anabolic effects of glucocorticoids may be ascribed to the
higher concentration of circulating androgens, which have stimulatory
effects on eating behavior and body weight gain [31]. In addition, it is
important to highlight that there was no difference in fluid intake
among the experimental groups of both protocol, indicating that the sex
differences on glucocorticoids-induced anabolic changes is not ascribed
to differences in fluid intake between males and females.
Additionally, the difference in corticosteroid receptors expression in
males and females may contribute to these differentiated GC responses
in both sexes. In fact, it was demonstrated that there is a higher
9
Physiology & Behavior 209 (2019) 112587
expression of corticosteroid receptors and the ability of GC to bind to
the hypothalamus and hippocampus in male animals than in females
[32,33]. In addition, pre-adipocytes from male animals have higher GR
expression and GC binding sites than females [34], which could justify
sex differences in the effects of GC on increasing visceral white adipose
tissues in males, but not in females, as observed in the present study.
Finally, it might be suggested that the anabolic effects of GC in male
rats, but not in females, are mediated by their actions in both corti-
costeroids receptors, GR and MR, since both glucocorticoids exerted
anabolic effects in males, and dexamethasone has more affinity to GR
while corticosterone can bind to both receptors [35].
In summary, the present study showed that the 28-day prolonged
treatment with dexamethasone was able to increase body weight, food
intake and weight and area of adipocytes of retroperitoneal adipose
tissue in males. The prolonged treatment of 28 days with corticosterone
stimulated body weight gain, food intake, weight and area of adipo-
cytes of perigonadal+perirenal adipose tissue and glucose intolerance
in males. However, these glucocorticoids had no effects in female rats.
Thus, these results suggest that males are more responsive to the ana-
bolic effects of prolonged treatment (28 days) with glucocorticoids than
females, probably due to the protective effect of circulating estrogens in
females, and/or the difference between the sexes in the expression/
activity of corticosteroid receptors in tissues evaluated in the present
study.
Acknowledgments
This work had the financial support of Conselho Nacional de
Desenvolvimento Cientifico e Tecnologico (CNPq) (Process: 422032/
2016-6) and Coordenação de Aperfeiçoamento de Pessoal de Nível
Superior (CAPES), Brazil.
Declaration of Competing Interest
The authors have nothing to disclose.
References
[1] M.W. Schwartz, S.C. Woods, D.J.R. Porte, R.J. Seeley, D.G. Baskin, Central nervous
system control of food intake, Nature 404 (2000) 661–671, https://doi.org/10.
1038/35007534.
[2] P.A. Tataranni, D.E. Larson, S. Snitker, J.B. Young, J.P. Flatt, E. Ravussin, Effects of
glucocorticoids on energy metabolism and food intake in humans, Am. J. Physiol.
Endocrinol. Metab. 271 (1996) 317–325, https://doi.org/10.1152/ajpendo.1996.
271.2.E317.
[3] B.S. McEwen, E.R. De Kloet, W. Rostene, Adrenal steroid receptors and actions in
the nervous system, Physiol. Rev. 66 (1986) 1121–1188, https://doi.org/10.1152/
physrev.1986.66.4.1121.
[4] C.J. Bautista, P.M. Martínez-Samayoa, E. Zambrano, Sex steroids regulation of ap-
petitive behavior, Mini Rev. Med. Chem. 12 (2012) 1107–1118, https://doi.org/10.
2174/138955712802762176.
[5] J. Hong, R.E. Stubbins, R.R. Smith, A.E. Harvey, N.P. Núñez, Differential suscept-
ibility to obesity between male, female and ovariectomized female mice, Nutr. J. 8
(2009) 1–5, https://doi.org/10.1186/1475-2891-8-11.
[6] C.F. de Souza, L.R.S. Stopa, G.F. Santos, L.C.N. Takasumi, A.B. Martins,
M.C. Garnica-Siqueira, R.N. Ferreira, F.G. de Andrade, C.M. Leite, D.A.M. Zaia,
C.T.B. Zaia, E.T. Uchoa, Estradiol protects against ovariectomy-induced suscept-
ibility to the anabolic effects of glucocorticoids in rats, Life Sci. 218 (2019)
185–196, https://doi.org/10.1016/j.lfs.2018.
[7] A. Santos Júnior, A.R. Carpinelli, R. Curi, The effect of controlled feeding conditions
on the metabolic characteristics of rats, Physiol. Behav. 45 (1989) 529–532,
https://doi.org/10.1016/0031-9384(89)90069-3.
[8] K. Veras, F.N. Almeida, R.T. Nachbar, D.S. de Jesus, J.P. Camporez, A.R. Carpinelli,
J.H. Goedecke, C.R. de Oliveira Carvalho, DHEA supplementation in ovar-
iectomized rats reduces impaired glucose- stimulated insulin secretion induced by a
high-fat diet, FEBS Open Bio 18 (2014) 141–146, https://doi.org/10.1016/j.fob.
2014.01.005.
[9] L.L. Bernardis, B.D. Patterson, Correlation between “Lee index” and carcass fat
L.R.S. Stopa, et al.
content in weanling and adult female rats with hypothalamic lesions, J. Endocrinol.
40 (1968) 527–528.
[10] K. Falholt, B. Lund, W. Falholt, An easy colorimetric micromethod for routine de-
termination of free fatty acids in plasma, Clin. Chim. Acta 46 (1973) 105–111,
https://doi.org/10.1016/0009-8981(73)90016-8.
[11] R. Guillemin, G.W. Clayton, J.D. Smith, H.S. Lipscomb, Measurement of free cor-
ticosteroids in rat plasma: physiological validation of a method, Endocrinology 63
(1958) 349–358.
[12] K.E. Zakrzewska, I. Cusin, A. Stricker-Krongrad, D. Boss, D. Ricquer, B. Jeanrenaud,
F. Rohner-Jeanrenaud, Induction of obesity and hyperleptinemia by central glu-
cocorticoid infusion in the rat, Diabetes 48 (1999) 365–375, https://doi.org/10.
2337/diabetes.48.2.365.
[13] I.N. Karatsoreos, S.M. Bhagat, N.P. Bowles, Z.M. Weil, D.W. Pfaff, B.S. McEwen,
Endocrine and physiological changes in response to chronic corticosterone: a po-
tential model of the metabolic syndrome in mouse, Endocrinology 151 (2010)
2117–2127, https://doi.org/10.1210/en.2009-1436.
[14] J.L. Steiner, M.E. Bardgett, L. Wolfgang, C.H. Lang, S.D. Stocker, Glucocorticoids
attenuate the central sympathoexcitatory actions of insulin, J. Neurophysiol. 112
(2014) 2597–2604, https://doi.org/10.1152/jn.00514.2014.
[15] C. Santos, F.B. Ferreira, L.M. Gonçalves-Neto, S.R. Taboga, A.C. Boschero,
A. Rafacho, Age- and gender-related changes in glucose homeostasis in gluco-
corticoid-treated rats, Can. J. Physiol. Pharmacol. 92 (2014) 867–878, https://doi.
org/10.1139/cjpp-2014-0259.
[16] L. Devenport, A. Knehans, A. Sundstrom, T. Thomas, Corticosterone's dual meta-
bolic actions, Life Sci. 45 (1989) 1389–1396, https://doi.org/10.1016/0024-
3205(89)90026-X.
[17] P. Björntorp, Do stress reactions cause abdominal obesity and comorbidities? Obes.
Rev. 2 (2001) 73–86, https://doi.org/10.1046/j.1467-789x.2001.00027.x.
[18] M. Rebuffé-Scrive, M. Brönnegard, A. Nilsson, J. Eldh, J.A. Gustafsson, P. Björntorp,
Steroid hormone receptors in human adipose tissues, J. Clin. Mol. Endocrinol. 71
(1990) 1215–1219, https://doi.org/10.1210/jcem-71-5-1215.
[19] P. Chimin, T.D. Farias, F.L. Torres-Leal, A. Bolsoni-Lopes, A.B. Campana,
S. Andreotti, F.B. Lima, Chronic glucocorticoid treatment enhances lipogenic ac-
tivity in visceral adipocytes of male Wistar rats, Acta Physiol. 211 (2014) 409–420,
https://doi.org/10.1111/apha.12226.
[20] J. Yu, B. Yu, J. He, P. Zheng, X. Mao, D. Chen, Chronic glucocorticoid exposure-
induced epididymal adiposity is associated with mitochondrial dysfunction in white
adipose tissue of male C57BL/6J mice, PLoS One 9 (11) (2014) e112628, https://
doi.org/10.1371/journal.pone.0112628.
[21] N.E. Block, M.G. Buse, Effects of hypercortisolemia and diabetes on skeletal muscle
insulin receptor function in vitro and in vivo, Am. J. Phys. 256 (1989) 39–48,
https://doi.org/10.1152/ajpendo.1989.256.1.E39.
[22] G. Dimitriadis, B. Leighton, M. Parry-Billings, S. Sasson, M. Young, U. Krause,
S. Bevan, T. Piva, G. Wegener, E.A. Newsholme, Effects of glucocorticoid excess on
the sensitivity of glucose transport and metabolism to insulin in rat skeletal muscle,
10
Physiology & Behavior 209 (2019) 112587
Biochem. J. 321 (1997) 707–712.
[23] H.C. Horner, A. Munck, G.E. Lienhard, Dexamethasone causes translocation of
glucose transporters from the plasma membrane to an intracellular site in human
fibroblasts, J. Biol. Chem. 262 (1987) 17696–17702.
[24] R. Nosadini, S. Del Prato, A. Tiengo, A. Valerio, M. Muggeo, G. Opocher,
F. Mantero, F. Duner, C. Marescotti, F. Mollo, F. Belloni, Insulin resistance in
Cushing’s syndrome, J. Clin. Endocrinol. Metab. 57 (1983) 529–536, https://doi.
org/10.1210/jcem-57-3-529.
[25] J.B. Ortiz, K.J. Mclaughlin, G.F. Hamilton, S.E. Baran, A.N. Campbell, C.D. Conrad,
Cholesterol and perhaps estradiol protect against corticosterone-induced hippo-
campal CA3 dendritic retraction in gonadectomized female and male rats,
Neuroscience 29 (2013) 409–421, https://doi.org/10.1016/j.neuroscience.2013.
04.027.
[26] E.A. Elbassuoni, Gender differences in ghrelin response to chronic immobilization
stress in rats: possible role of estrogen, Gen. Physiol. Biophys. 33 (2014) 111–120,
https://doi.org/10.4149/gpb_2013061.
[27] L. Zhu, M.N. Martinez, C.H. Emfinger, B.T. Palmisano, J.M. Stafford, Estrogen
signaling prevents diet-induced hepatic insulin resistance in male mice with obe-
sity, Am. J. Physiol. Endocrinol. Metab. 306 (2014) 1188–1197, https://doi.org/10.
1152/ajpendo.00579.2013.
[28] S.A. Litwak, J.L. Wilson, W. Chen, C. Garcia-Rudaz, M. Khaksari, M.A. Cowley,
P.J. Enriori, Estradiol prevents fat accumulation and overcomes leptin resistance in
female high-fat diet mice, Endocrinology 155 (2014) 4447–4460, https://doi.org/
10.1210/en.2014-1342.
[29] E. Riant, A. Waget, H. Cogo, J.F. Arnal, R. Burcelin, P. Gourdy, Estrogens protect
against high-fat diet-induced insulin resistance and glucose intolerance in mice,
Endocrinology 150 (2009) 2109–2117, https://doi.org/10.1210/en.2008-0971.
[30] R. Matysková, B. Zelezná, J. Maixnerová, D. Koutová, M. Haluzík, L. Maletínská,
Estradiol supplementation helps overcome central leptin resistance of ovar-
iectomized mice on a high fat diet, Horm. Metab. Res. 42 (2010) 182–186, https://
doi.org/10.1055/s-0029-1243250.
[31] L. Asarian, N. Geary, Sex differences in the physiology of eating, Am. J. Physiol.
Regul. Integr. Comp. Physiol. 305 (2013) R1215–RE126, https://doi.org/10.1152/
ajpregu.00446.2012.
[32] D. Karandrea, C. Kittas, E. Kitraki, Contribution of sex and cellular context in the
regulation of brain corticosteroid receptors following restraint stress,
Neuroendocrinology 71 (2000) 343–353, https://doi.org/10.1159/000054555.
[33] I. Elaković, A. Djordjevic, M. Adzic, J. Djordjevic, M. Radojčić, G. Matić, Gender-
specific response of brain corticosteroid receptors to stress and fluoxetine, Brain
Res. Rev. 1384 (2011) 61–68, https://doi.org/10.1016/j.brainres.2011.01.078.
[34] J.M. Joyner, L.J. Hutley, D.P. Cameron, Glucocorticoid receptors in human pre-
adipocytes: regional and gender differences, J. Endocrinol. 166 (2000) 145–152.
[35] D.L. Tempel, S.F. Leibowitz, Adrenal steroid receptors: interactions with brain
neuropeptide systems in relation to nutrient intake and metabolism, J.
Neuroendocrinol. 6 (1994) 479–501.