Arsenic-induced testicular toxicity in Gallus gallus: Expressions
of inflammatory cytokines and heat shock proteins
Xiao Sun,2 Siwen Li,2 Ying He, Hongjing Zhao, Yu Wang, Xiangwei Zeng,1 and Mingwei Xing1
College of Wildlife Resources, Northeast Forestry University, Harbin 150040, Heilongjiang Province, China
ABSTRACT The aim of the study is to investigate
the effects of sub-chronic poisoning with arsenic on
the testes of chickens treated. Seventy-two 1-day-old
chickens were randomly divided into 4 groups and pro-
vided food with different doses arsenic. The histological
changes were examined. The mRNA levels of inflam-
matory factors, including transcription factor nuclear
factor-κB (NF-κB), tumor necrosis factor-α (TNF-α),
inducible nitric oxide synthase (iNOS), cyclooxygenase-
2 (COX-2), prostaglandin E synthase (PTGEs), and
heat shock proteins (Hsps), including Hsp70, Hsp90,
Key words: Arsenic, Chicken, Testicle, Inflammatory factor, Heat shock protein
INTRODUCTION
Arsenic is one of the most common toxins and a ma-
jor health concern globally due to its wide distribution
and adverse health effects. It is widely used as a feed
additive in the production of animals at present. Ar-
senic has been proposed as a cause of reproductive
failure in male workers at a copper smelter in Swe-
den, but this suggestion should be viewed with cau-
tion, as subjects were also exposed to other potentially
hazardous metals (Beckman, 1978). Nevertheless, ar-
senic is considered to be a likely reproductive toxicant
that might cause male-mediated reproductive effects
(Hopenhayn-Rich et al., 1999). In the epididymides
of mice, the presence of radioactive arsenic indicated
a risk of decreasing sperm viability and impaired
reproduction (Danielson et al., 1984; Pant et al., 2001).
Furthermore, it was reported that arsenic exposure pro-
duced steroidogenic dysfunction causing impairment of
spermatogenesis in rats (Sarkar et al., 2003).
Arsenic exposure influences the expression of inflam-
matory cytokines in the airway of women (Dutta et al.,
2015). Inflammation is a defensive reaction to injury
factors of the vascular system in living tissue. Many
inflammatory cytokines are involved in the inflamma-
tory response: for instance, the transcription factor nu-

C 2017 Poultry Science Association Inc.
Received November 30, 2016.
Accepted March 15, 2017.
1
Corresponding author: xingmingwei@nefu.edu.cn
2
Xiao Sun and Siwen Li contributed equally to this work.
1
Hsp60, Hsp40, and Hsp27 were assessed by quantita-
tive real-time PCR in the testes of chickens. The protein
expressions of iNOS, Hsp60, and Hsp70 were detected
by western blot. Increased mRNA and protein levels of
inflammatory factors and Hsps with testicular damage
showed that arsenic-induced testicular toxicity includes
inflammatory and heat shock response in chickens, and
that increased Hsps levels may play a protective role in
inflammation damage induced by arsenic on the testes
of chickens.
2017 Poultry Science 0:1–8
http://dx.doi.org/10.3382/ps/pex073
clear factor-κB (NF-κB) can regulate the expressions
of okadaic acid, a phosphatase 1 and 2A inhibitor in-
volved in the inflammatory response (Schmidt et al.,
1995). After activation of NF-κB, it can regulate the
expression of a series of genes, including tumor necrosis
factor-α (TNF-α) and inducible nitric oxide synthase
(iNOS), among others (Schmidt et al., 1995). TNF-α
is an important pro-inflammatory cytokine mainly pro-
duced by a series of immune cells, including T lympho-
cytes, B lymphocytes, natural killer cells, eosinophils,
basophils, neutrophils, and monocytes (Varfolomeev
and Ashkenazi, 2004). It can also be generated by some
non-immune cells, including Kupffer cells, fibroblasts,
smooth muscle cells, endothelial cells, glial cells, and
neurons (Thomson and Lotze, 2003). Moreover, TNF-
α can produce calcium-independent iNOS, release NO,
and cause tissue inflammation (Blanco et al., 2007). En-
zymes of iNOS and cyclooxygenase-2 (COX-2), which
are involved in tissue damage, can induce prostaglandin
E synthase (PTGEs; Surh et al., 2001). Therefore, re-
search on these inflammatory cytokines in the testes
of chickens treated with arsenic trioxide (As2 O3 ) may
be advantageous to understand whether heavy metal
arsenic induces an inflammatory response.
Inflammatory cytokines can induce heat shock pro-
teins (Hsps) upon infection with a virus (Phillips et al.,
1991). In 1962, Ritossa observed that the polytene chro-
mosomes of normal flies showed “puffing”; later, the
transcription of those puffs were strengthened under
high temperature, which may prompt some kind of Hsps
synthesis (Tissieres et al., 1974; Ritossa, 1996). Hsps are
2 SUN ET AL.
a group of proteins that are generated by environmen-
tal stressors under high temperatures (Del et al., 2001).
Hsps are highly conserved during the process of evolu-
tion, such that the Hsps homology of bacteria and hu-
mans shares more than 50% (Craig, 1985). Among these
proteins, Hsp70 is the most important Hsps (Hunt and
Morimoto, 1985). To avoid aggregation, Hsp70 is able
to recognize and bind to hydrophobic groups around
them before folding the precursor protein. Its function
is to remit cell injuries and enhance the body’s resis-
tance to diverse stressors (Kang et al., 1990). Hsp90
is an important molecular chaperone that participates
in the synthesis of many key molecules within the cell
(especially those regulating cell growth and the sur-
vival of many signaling molecules), affecting the sta-
bility and function of these molecules (Neckers and Ivy,
2003). Hsp60 synthesis is stimulated by protein denatu-
ration (Hightower, 1993). Hsp40 and Hsp27 are mainly
involved in the actin stability and cytokine signal trans-
duction, protecting cells from damage of various stress
factors (Mehlen et al., 1996).
To the best of our knowledge, few studies have ex-
amined the expression of inflammatory cytokines and
heat shock proteins in relation to arsenic-induced tes-
ticular toxicity. The present study was performed to
investigate the protective effects of inflammatory cy-
tokines and heat shock proteins on testicular toxicity
using chicken as a model.
MATERIALS AND METHODS
Animal Care and Drug Treatment
All procedures used in this study were approved by
the Institutional Animal Care and Use Committee of
Northeast Forestry University (Harbin, China) (UT-31;
20 June 2014). The highest dose of sub-chronic tox-
icity test can use 1/20 to 1/5 of the median lethal
dose (LD50 ) [Appendix 1], and LD50 of arsenic for
chicken was 50 mg/kg BW [Appendix 2]. Base on that,
72 1-day-old chickens were randomly divided into 4
groups: the low arsenic dose group (L group, contain-
ing 7.5 mg/kg As2 O3 ), the medium arsenic dose group
(M group, containing 15 mg/kg As2 O3 ), the high
arsenic dose group (H group, containing 30 mg/kg
As2 O3 ), and the control group (C group, containing
0 mg/kg As2 O3 ). As2 O3 was added into the food to
make supplements uniformed according to the chicken
LD50 of As2 O3 . Each group contained 18 chickens. The
experimental animals were provided adequate food and
water. Each chicken was fed with the same amount of
feed, and detected the changes of body weight.
The composition of the diet was 2.766% metabo-
lizable energy (kcal/kg), 18.45% crude protein, 2.82%
crude fiber, 3.80% calcium, and 0.62% phosphorus,
which had been added to the feedstuff to meet the
minimum requirements for energy and nutrients for
chickens and without influencing subsequent euthanasia
with sodium pentobarbital (National Research Council,
1994).
Euthanasia and Testes Collection
Animals were anesthetized and euthanized by injec-
tion of sodium pentobarbital (30 mg/kg body weight),
and the testes of the chickens removed quickly on
days 30, 60, and 90, respectively. Then the testis tis-
sue specimens were fixed with 2.5% glutaraldehyde in
0.1 M sodium phosphate buffer for electron microscopy,
and the other samples were directly stored at −80◦ C
until further use.
Ultrastructural Observations
For electron microscopy, testis tissue specimens were
fixed with 2.5% glutaraldehyde in 0.1 M sodium phos-
phate buffer (pH 7.2) for 3 h at 4◦ C, washed in the
same buffer for 1 h at 4◦ C and post-fixed with 1% os-
mium tetroxide in sodium phosphate buffer for 1 h at
4◦ C. The tissues were then dehydrated in a graded se-
ries of ethanol starting at 50% ethanol for 10 min at a
time and then were immersed twice in propylene oxide.
The tissue specimens were mounted using a 10:1 ratio
of epoxy (Araldite GY-502 Epoxy, Huntsman Interna-
tional LLC, Salt Lake City, UT): hardener (Hysol HD
3416 Hardener, Loctite/Henkel Corp., Atlanta, GA).
Ultrathin sections were stained with Mg-uranyl ac-
etate and lead citrate for evaluation, and testis samples
were observed by a transmission electron microscope
(Xing et al., 2015).
The mRNA Levels of Inflammatory
Cytokines and Hsps in the Testes
of Chickens Exposure to As2 O3
Specific primers for inflammatory cytokines (NF-κB,
TNF-α, iNOS, COX-2, PTGEs), Hsps (Hsp70, Hsp90,
Hsp27, Hsp40, Hsp60), and the internal reference of
glyceraldehyde-3-phosphate dehydrogenase (GAPDH)
were designed by the Primer Premier software (PRE-
MIER Biosoft International, Palo Alto, CA) referring
to known chicken sequences (Appendix 3).
Total RNA was extracted from the testes (50 mg tis-
sue, n = 6/group) using RNAiso Plus reagent (Takara,
China). The concentration and purity of the total
RNA were determined spectrophotometrically at the
absorbance of OD260/280 nm (Gene Quant 1300/100,
USA). The RNA was reverse-transcribed to cDNA us-
ing the PrimeScript RT Reagent Kit (Takara, China)
according to the manufacturer’s instructions. Synthe-
sized cDNA was diluted 10 times with sterile, double-
distilled water and stored at −80◦ C before use.
Quantitative real-time PCR was completed using
the FastStart Universal SYBR Green Master reagents
(Roche USA, Branford CT) on a BIOER LineGene
9600 sequence detection system (BIOER, China). Each
 ARSENIC INDUCES CHICKEN TESTICULAR TOXICITY
Figure 1. Ultrastructural changes of chickens’ testes tissue sections. A and F represented testes of chickens from Control group. B and G
represented testes of chickens from L groups. C and H represented testes of chickens from M group. D, E, and I represented testes of chickens
from H groups at 90 days.
sample was evaluated in triplicate. The detailed con-
ditions of PCR protocol and the method of the rela-
tive mRNA abundance on the basis of the method of
2−ΔΔCt were indicated in our previous research (Leite
et al., 2013; Guo et al., 2016).
Western Blot Analysis of iNOS, Hsp60,
and Hsp70
All of the 50 mg of testis tissue for each sample were
rinsed in saline and then homogenized in sodium do-
decyl sulfate (SDS) Lysis Buffer (Beyotime, China),
which contained 1 mM phenylmethylsulfonyl fluoride.
After centrifuging, the supernatants were collected. The
concentrations of protein were detected using BCA pro-
tein assay kits to equal the protein extracts doses of all
groups.
The protein extracts were separated by 12% SDS-
polyacrylamide gel electrophoresis. The separated pro-
teins were transferred to a polyvinylidene fluoride
(PVDF) membrane, then blocked for 2 h at 37◦ C
and incubated with rabbit anti-chicken iNOS, Hsp60 or
Hsp70 polyclonal antibodies (1:1000, provided by Prof.
Xu, Northeast Agricultural University), and GAPDH
antibody (1:1000, Beyotime, China) overnight at 4◦ C.
After being washed 3 times with Tris-buffered NaCl
solution with tween 20 (TBST), the membrane was
then incubated with a horseradish peroxidase (HRP)-
labeled goat anti-rabbit IgG (1:10000, Beijing Biosyn-
thesis Biotechnology Co., LTD, China) for 1 h at 37◦ C.
By using the enhanced chemiluminescence (ECL) west-
ern blotting detection kit (Thermo Scientific, Waltham,
MA), the protein expression bands were visualized, and
then the membrane was exposed to an X-ray film. The
GAPDH signal was used as an internal reference pro-
tein standardization to obtain relative amounts of the
proteins of each sample.
3
Statistical Analyses
All statistical analyses of the data were performed
using SPSS 13.0 software. The level of statistical about
significant differences value (P < 0.05) was obtained by
one-way analysis of variance (ANOVA). Data regard-
ing differences between the mean values of normally dis-
tributed data were assessed by Tukey’s honestly signif-
icant difference test for post hoc multiple comparisons.
Data were expressed as the mean ± SD of 6 observa-
tions.
RESULTS
Ultrastructural Changes
Testes tissues from the control group showed nor-
mal ultrastructure structures with regular cells on
90 days (Figure 1A and 1F). In the corresponding L,
M, H group, arsenic treatment caused extensive injury
of the testis tissues. The mitochondria in testis tissues
of the arsenic groups were swollen and vacuolated with
degeneration or loss of cristae. Cells showed typical
chromatin condensation and margination and shrink-
age of their nuclei comparison with the control group
(Figure 1B-E and 1G-I), and there was apoptosis with
perinuclear chromatin condensation in H group. The
alteration degree was in a dose-dependent manner.
Effects of As2 O3 on the mRNA Levels
of NF-κB, TNF-α, iNOS, COX-2, and PTGEs
in the Testes of Chickens
The effects of As2 O3 on the mRNA levels of in-
flammatory cytokines in testes of chickens were shown
in Figure 2. The NF-κB and TNF-α mRNA levels of
testes tissues were significantly increased (P < 0.05 or
4 SUN ET AL.

Figure 2. Effects of As2 O3 on the mRNA levels of NF-κ B, TNF-α , iNOS, COX-2, and PTGEs in the testes of chickens. A represented the
mRNA levels of NF-κ B in the testes. B represented the mRNA levels of TNF-α in the testes. C represented the mRNA levels of iNOS in the
testes. D represented the mRNA levels of COX-2 in the testes. E represented the mRNA levels of PTGEs in the testes. Each value represented
the mean ± SD of 6 individuals (n = 6). The asterisk or double asterisk indicates that there were significant differences (∗ P < 0.05 or ∗∗ P <
0.01) between the C group and L, M, H group at the same time point, respectively.

P < 0.01) in arsenic treatment groups compared with
control groups, except for the L group on days 30 with
no significant changes (Figure 2A and B). Compared
with control groups, the iNOS, COX-2, and PTGEs
mRNA levels of testes tissues were significantly in-
creased (P < 0.05 or P < 0.01) (Figure 2A-E).
Effects of As2 O3 on the mRNA Levels
of Hsp70, Hsp90, Hsp60, Hsp40, and Hsp27
in the Testes of Chickens
Effects of As2 O3 on the mRNA levels of Hsp70,
Hsp90, Hsp60, Hsp40, and Hsp27 in testes of chickens
were shown in Figure 3. The mRNA levels of Hsp70,
Hsp90, Hsp40, and Hsp27 in the testes were shown in
Figure 3A, B, D and E. The L, M, and H groups on
days 30, 60, and 90 had a significant increase (P < 0.05
or P < 0.01) compared with those in the correspond-
ing control groups. The mRNA levels of Hsp60 in the
testes were shown in Figure 3C. Compared with the
control group, the M and H group on days 30, 60, and
90 had a significantly increase (P < 0.05 or P < 0.01).
From the information above, the Hsp70, Hsp90, Hsp60,
Hsp40, and Hsp27 mRNA transcription increased in a
dose-dependent manner (P < 0.05 or P < 0.01).
Western Blot Analysis of iNOS Protein
Expressions in the Testes of Chickens
In order to investigate the effects of arsenic-toxicity
on inflammation-related protein expressions in testes
of chickens, iNOS expressions were examined by West-
ern blot and showed in Figure 4. Compared with the
corresponding control groups, the protein expressions
of iNOS in the chronic arsenic treatment group for
iNOS increased significantly (P < 0.05 or P < 0.01)
(Figure 4B).
Western Blot Analysis of Hsp60 and Hsp70
Protein Expressions in the Testes of
Chickens
In order to investigate the effects of arsenic-toxicity
on Hsps levels in testes of chickens, Hsp60 and Hsp70
expressions were examined by Western blots. The pro-
tein expressions of the H groups for Hsp60 in the testes
 ARSENIC INDUCES CHICKEN TESTICULAR TOXICITY 5

Figure 3. Effects of As2 O3 on the mRNA levels of Hsps in the testes of chickens. A represented the mRNA levels of Hsp70 in the testes. B
represented the mRNA levels of Hsp90 in the testes. C represented the mRNA levels of Hsp60 in the testes. D represented the mRNA levels of
Hsp40 in the testes. E represented the mRNA levels of Hsp27 in the testes. Each value represented the mean ± SD of 6 individuals (n = 6). The
asterisk or double asterisk indicates that there were significant differences (∗ P < 0.05 or ∗∗ P < 0.01) between the C group and L, M, H group at
the same time point, respectively.

in all groups increased significantly on days 30 and 90
(P < 0.05 or P < 0.01) in comparison to the correspond-
ing control groups (Figure 5B). However, the protein
expressions of the H group for Hsp60 were decreased
significantly on days 60 (P < 0.05 or P < 0.01) (Fig-
ure 5B). Nevertheless, the protein expressions of the
H group for Hsp70 was increased significantly in com-
parison to the control (P < 0.05 or P < 0.01) (Fig-
ure 5C), except for the H group on days 60 with no
significant changes.
DISCUSSION
Arsenic, a heavy metal, is toxic when it penetrates
into human and animal bodies by inhalation (Dutta
et al., 2015). Following arsenic exposure, the matu-
ration of spermatogonia through the process of meio-
sis has been reported to be severely disrupted (Omura
et al., 2000; Sanghamitra et al., 2007). Tissues are dam-
aged following the change of the expression of inflam-
matory cytokines and heat shock proteins (Zhao et al.,
2013). For the purpose of detecting arsenic-induced tis-
sues damage, we examined the histological (Appendix
4, unpublished data) and ultrastructural observations,
which showed testicular damage with inflammatory
cell infiltration, typical chromatin condensation and
margination, and shrinkage of their nuclei in compari-
son with the control group. Results indicated that the
tissues of chickens treated with arsenic were damaged
in a time- and dose-dependent manner compared with
the control group. We found arsenic ingestion reduced
chicken body weight slightly; however, the testes weight
tended to be reduced significantly (Appendix 5, unpub-
lished data), which was similar with Adedara’s study
(Adedara et al., 2017). In addition, the heart induced
by chronic arsenic toxicity was reported to be partially
reversible after cessation of As2 O3 administration (Wu
et al., 2003).
We examined the inflammatory reactions, such as the
mRNA levels of NF-κB, TNF-α, iNOS, COX-2, and
PTGEs on the testes by quantitative real-time PCR
(qPCR). Based on our results, we observed that the
NF-κB, TNF-α, iNOS increased in a time- and dose-
dependent manner. COX-2 and PTGEs mRNA levels
of testes tissue were significantly increased, suggest-
ing that both of them may play influential roles in
tissue inflammation injury induced by arsenic toxic-
ity within the testes. Previous studies have reported
the similar findings that arsenic toxicity could disrupt
the homeostasis and cause inflammation injury to the
6 SUN ET AL.

gastrointestinal tract of chickens (Xing et al., 2015). In

Figure 4. Effects of As2 O3 on proteins expressions of iNOS in the
testes of chickens. A represented the protein expressions of iNOS in
the testes. C30 , C60 , and C9 0 represented C groups at 30, 60, and
90 d, respectively. H30 , H60 , and H90 represented H groups at 30, 60,
and 90 d, respectively. B represented iNOS/GAPDH ratio in the testes.
The values presented the means ± SD of three individuals (n = 3).
The asterisk or double asterisk indicates that there were significant
differences (∗ P < 0.05 or ∗∗ P < 0.01) between the C group and H
group at the same time point, respectively.
addition, lead induced inflammatory damage in testes
of Swiss albino mice (Barbhuiya et al., 2013), and
the mRNA level of TNF-α in the zebrafish larvae
treated with copper increased in a time-dependent man-
ner (Leite et al., 2013). Results were consistent with
the previous conclusion that PTGEs catalyzed the iso-
merization of PGH2 to PGE2, and the latter played
an important role in inflammation (Liu et al., 2012),
cell growth, and transformation (Martin et al., 1991).
TNF-α induces the activation of NF-κB in the COX-2
promoter, followed by initiation of COX-2 expression
(Chen et al., 2000). Our results were similar to a prior
study by Fu et al. (2013), which showed that expression
of COX-2 and PTGEs in the intestinal tract were in-
creased by cold stress at early time points, but reduced
after 20 d of cold exposure. Our results showed that
after adding As2 O3 to the food, COX-2 mRNA levels
in M and H groups increased on days 30 and 60 com-
pared with the corresponding control groups; however,
these declined on d 90, so we surmised that TNF-α in-
duced the activation of COX-2. Compared with control
group, our results revealed that excess arsenic could up-
regulate NF-κB, TNF-α, iNOS, COX-2, and PTGEs
mRNA levels.
The mRNA expression levels of Hsps were up-
regulated compared with control groups; as molecular
chaperones, Hsps are synthesized by a group of spe-

Figure 5. Effects of As2 O3 on the protein expressions of Hsp60, Hsp70 in the testes of chickens. A represented the protein expressions of
Hsp60, Hsp70 and GAPDH in the testes. C30 , C60 , and C90 represented C groups at 30, 60, and 90 d, respectively. H30 , H60 , and H90 represented
H groups at 30, 60, and 90 d, respectively. B represented Hsp60/GAPDH ratio in the testes. C represented Hsp70/GAPDH ratio in the testes.
The values presented the means ± SD of three individuals (n = 3). The asterisk or double asterisk indicates that there were significant differences
(∗ P < 0.05 or ∗∗ P < 0.01) between the C group and H group at the same time point, respectively.
 ARSENIC INDUCES CHICKEN TESTICULAR TOXICITY
cific protein when under stress, such as high fever,
oxidative stress, infections, nutritional deficiencies, or
exposure to harmful chemicals and inflammatory cy-
tokines (Lindquist and Craig, 1988; Yao et al., 2013a,b).
In our study, the increased inflammation factors sug-
gested that an inflammatory response in the testes was
induced by As2 O3 , which might lead to testicular in-
jury. Increasing numbers of research studies in the lit-
erature support the important roles of Hsps in the in-
flammatory response. Hsps are involved in signal trans-
duction and controlling of cytokine genes to enhance
the role of T lymphocyte antigen presentation, partic-
ipating in cell proliferation and the mechanism caused
by inflammation (Suzuki et al., 1991). Previous studies
have shown that As2 O3 increased mRNA expressions
of Hsp27, Hsp40, Hsp60, Hsp70, and Hsp90 in chicken
immune organs (Guo et al., 2016). Moreover, arsenite
could stimulate Hsp70 expression in salmon (Grosvik
and Goksoyr, 1996). Cao et al. reported that heavy
metal (Mo, Ca) exposure raised the mRNA expressions
of Hsp60 and Hsp70, and there was a positive correla-
tion between metal concentration and expressions (Cao
et al., 2016). Thus, in our study, the increased inflam-
mation factors suggested that the testes inflammatory
response was induced by As2 O3 , which might lead to
testicular injury. Quite notably, in this study, mRNA
expression levels of Hsps were up-regulated comparing
with control groups, indicating that Hsps in testes were
overexpressed because of stress caused by heavy met-
als and were synthesized in large quantities to protect
interstitial cell from damage through the Hsp path-
way. These results were also consistent with the study
by Rajeshkumar and Munuswamy (2011), which re-
ported that heavy metals such as Ag, Cd, and Cu could
increase the Hsps expression in milk fish.
Expression of genes in the Hsp70 family may be
related to changes in pro-inflammatory cytokines in-
duced by exposure to As2 O3 (Sreedhar and Csermely,
2004). By activation of phospholipase A2 (PLA2) and
promotion the rapid increase in intracellular ROS,
TNF-α caused cellular lipid peroxidation and resulted
in inflammatory tissue damage (Jacquier-Sarlin et al.,
1994). Hsps play roles in protecting cells and tissues
against inflammatory damage by inhibiting cytokines
(Coelho et al., 2008). The previous results portray pos-
itive correlations between the serum levels of Hsp70
and various markers of inflammation in elderly pa-
tients (Njemini et al., 2004), which were in accord with
the current study, the expression of TNF-α and Hsps
showed a similar trend (Figure 2A and 3A). However,
it is suggested that iNOS could induce the generation
of Hsp70, astrocytes treated by LPS and TNF-α could
induce iNOS production and increase the protein ex-
pressions of Hsp70 stress protein, both of which were
similar with the experiment (Calabrese et al., 2000).
Furthermore, the results of PTGEs mRNA levels
showed that when the body began to heat shock reac-
tion, Hsps mRNA expressions significantly increased,
but PTGEs mRNA expression had a slowing growth
7
rate, suggesting that PTGEs might be inhibited by
Hsps. This is consistent with our results.
Arsenic causes development of various hazardous ef-
fects on testes of chickens and induces testicular toxi-
city. Inflammatory and heat shock response were trig-
gered and might play protective roles in inflammation
damage. Results of the present study strongly suggested
that exposure to arsenic might have adverse effects on
testicular development. However, so far, the specific
As2 O3 -activated induction and regulation mechanisms
among Hsps and inflammatory cytokines are unclear,
and further studies are needed.
CONCLUSION
In conclusion, the present experiments showed that
arsenic induced testicular toxicity in chickens. Arsenic
exposure caused inflammatory and heat shock response,
and the increased of Hsps levels may play a protec-
tive role in inflammation damage induced by As2 O3 in
chicken testes.
ACKNOWLEDGMENTS
This study was supported by the National Natural
Science Foundation of China (Grant No. 31672619),
the Fundamental Research Funds for the Central Uni-
versities (Grant No. 2572016EAJ5) and the Natural
Science Foundation of Heilongjiang Province (Grant
No. C2015061).
Conflict of interest: No author has any financial or per-
sonal relationships that could inappropriately influence
or bias the dose of the paper.
SUPPLEMENTARY DATA
Supplementary data are available at PSCIEN online.
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