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THE EFFECT OF TRAMADOL TREATMENT ON RAT TESTES AND THE POSSIBLE

PROTECTIVE ROLE OF SELENIUM (LIGHT AND ELECTRON MICROSCOPIC
STUDY)

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 AL-AZHAR ASSIUT MEDICAL JOURNAL VOL 13 , NO 1 , JANUREY 2015 SUPPL

THE EFFECT OF TRAMADOL TREATMENT ON RAT TESTES AND THE

POSSIBLE PROTECTIVE ROLE OF SELENIUM (LIGHT AND ELECTRON
MICROSCOPIC STUDY)
Eman Badawi El Shal* and Mai M.H. Selim**
Anatomy* and Histology** departments –Faculty of Medicine for Girls Al-Azhar University
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ABSTRACT
Introduction: Pain is a major health problem. Opioid drugs remain the first choice for relief of
moderate to severe pain conditions. Tramadol (Tr) is a centrally acting opioid which decreased semen
quality in chronic tramadol users. On the other hand, selenium (Se) was found to be antioxidant
nutrient that is essential for normal testicular development. Aim of the work: the current work was
aimed to study the light and electron microscopic changes occurring in rat testes following tramadol
administration and evaluate the role of selenium. Materials and methods: forty adult male albino rats
were divided into 4 equal groups: group I (control group), group II (Se group) which received a daily
dose of Se (31.5 microgram/rat/day) dissolved in distilled water (D.W.). Group III; (Tr group): the
animals received a daily dose of Tr (7mg/rat/day) dissolved in D.W. Group VI; (Se and Tr group): in
this group the animals were given both Se and Trof the same mentioned doses. All doses were given
orally by orogastric tube for 8 weeks. Testicular samples were prepared for light and electron
microscopic examination. Results:The germinal epitheliumof the seminiferous tubules of the tramadol
group showed that, the spermatogenic cells were replaced by vacuoles or acidophilic materials.
Spermatids and sperms were rarely seen. Ultrastructurally, some of the spermatogenic cells were seen
ruptured, and contained small dark condensed nuclei. The basement membrane (B.M.) was
discontinuous at certain pointsand the collagen fibers were detected around it.Some Leydig cells were
seen ruptured. Administration of a concomitant dose of Se and Tr improved all the previously
mentioned changes. Conclusion:In adult albino rat tramadol induced damage of the testicular
structure, which was ameliorated by the concomitant Se administration.
Key words: testes, tramadol, selenium, histology, oxidative stress, antioxidants.
INTRODUCTION Selenium (Se) has received considerable
Painis a major health problem in attention as an essential micronutrient for both
medicine. Despite the current introduction of animal and human beings (Ali et al., 2011). It
many new analgesics, opioid drugs remain the has an important role in the regulation of the
first choice to relief moderate to severe pain reproductive system and plays a role in male
condition, which require long term treatment infertility (Swathy et al.,2006).
(Ceccarelli et al.,2006). Recently, it was found Selenium is an important antioxidant
that intrathecal and oral opioids are capable of nutrient. It is essential for normal testicular
suppressing testosterone secretion throughout development and spermatozoa motility and
their period of administration (Aloisiet al., functions (Messaoudi et al., 2010). Ursini et al.
2009; Aloisi et al., 2011). Men consuming (1999) reported that supplementation of Se in
therapeutic opioids several times daily for sub fertile men with low Se status could
treatment of chronic pain frequently develop improve sperm motility and increase the chance
erectile dysfunction, decreased libido, fatigue of successful conception. Also, it has been
and depression (Ahmed and Kurkar,2014). demonstrated to have the protective effects
Tramadol (Tr) is a centrally acting opioid against the toxicity of chemicals andmetals in
analgesic which is mainly used for the treatment the male reproductive system of experimental
of moderate to severe pain (Nossaman et al., animals(Fahmy et al., 2008;Said et al., 2010)
2010).Tr causes respiratory depression, This study was designed to evaluate the
psychological and physical addiction similar to effect of long term tramadol treatment on the
that of other opiates. Repeated Tr administration testicular structure in adult male albino rats and
might lead to accumulation of toxic metabolites the possible protective role of selenium.
in the patient's bodies increase risk for MATERIAL AND METHODS
pharmacokinetic interaction and/or increasing 40 adult male albino rats(180-200 gm)
its potential for toxicity (Abou El Fatoh et al., were used. The animals were purchased from
2014).Decreased semen quality was also, noted the Egyptian organization for biological
in chronic Tr users (Ahmed and Kurkar, products and vaccines (Cairo, Egypt), they were
2014). kept in clean properly ventilated cages, in the
animal house of anatomy department, Faculty of
126 | P a g e
 Eman Badawi El Shal* and Mai M.H. Selim**
medicine for girls Al-AzharUniversity. Free
access to food and water was allowed
throughout the experiment. They were left for
one week before the experiment for
acclimatization.
The animals were randomly divided into
4 equal groups:
Group I (G I):were used as a control
group, divided into 2 subgroups; five of them
were given 0.7ml of distilled water /day by
orogastric tube, while the other 5 were left
without any treatment.
Group II (G II)(Se group):selenium was
purchased from (Wassen Company, UK) in the
form of tablets; each tablet contains 100
microgram, the equivalent human therapeutic
dose was 31.5 microgram/rat/day. This dose
was dissolved in 0.6 ml of D.W. given to the
animal by orogastric tube(Koyuturk et al.,
2007).
Group III(G III)(Tr group): tramadol
was in the form of capsules, each one contained
100mg of tramadol hydrochloride. The Tr dose
was calculated according to (Paget and Barnes,
1964) and according to the maximum human
therapeutic daily dose which was 400mg
(Brunton et al., 2006). One capsule was
evacuated and dissolved in 100 ml of D. W. to
obtain a concentration of 10mg/1ml D.W. given
daily to the animal by orogastric tube(0.7ml/
rat).TheTrwas purchased from local pharmacy.
Group IV (G IV)(SeandTr group): in
this group the animals were given both Se and
Tr daily by the previously mentioned doses and
route of administration, one quarter of hour
space between each drug.
The experiment continued for 8 weeks
then the animals were exposed to ether
inhalation, both testes were taken from the
scrotal sac, collected from all experimental
groups.The left testis was fixed inBouin's
solution(Drury and Wallington, 1980; Carson
and Hladik, 2009) processed for paraffin
blocks formation, serial sections of 5 Micron
thickness were cut and stained with hematoxylin
and eosin (H &E) for routine histological
examination (Kiernan, 2001). The slides were
examined under the light microscope; the
images were taken by a microscope connected
to a digital camera.
For electron microscopic examination the
right testis of each rat was immediately
dissected into a very small pieces of 1x1 mm³
and fixed in 3% phosphate buffered
gluteraldehyde for 24 hours and post fixed in
1% osmium tetraoxide in the same buffer for 1.5
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hour. The testes were further processed to form
ultrathin sections. The ultrathin sections were
cut, and double stained with urinyl acetate and
lead citrate (Hajibagheri,1999). Where, it was
ready for examination by Joel jem transmission
electron microscope at 60 KV accelerating
voltage. The electron microscopic study was
done at the Regional center for mycology and
Biotechnology AL-Azhar University.
RESULTS
Light microscopic results
Examination of H &E stained control
testicular sections of both subgroups were
completely similar, showed that the testis was
covered by C.T. capsule, tunica albuginea. The
parenchyma was made up of many seminiferous
tubules cut in various planes. The seminiferous
tubules were closely approximated with each
other, few interstitial tissue was present in
between themfig (1).
The seminiferous tubules were lined by
stratified epithelium; which was of two types,
the spermatogenic cells in various stages of
growth and non spermatogenic the supportive
larger Sertoli cells. Many spermatogenic cells
could be recognized as spermatogonia, they
were found in the basal layer of seminiferous
tubule. They characterized by rounded or oval
nuclei with condensed chromatin. 1ry
spermatocytes were characterized by their
copious cytoplasm and large nuclei containing
coarse clumps or thin threads of chromatin. The
spermatids could be recognized by its small
rounded darkly stained nuclei and its position
towards the luminal end while spermatozoa
could be recognized by their elongated shape
and pointed end fig (2).
Sertoli cells were the supportive cells that
present in an intimate relation to the
spermatogenic cells. They were rested on the
basement membrane of the seminiferous tubule
and extended to the lumen of the tubule, the
nuclei of these cells were oval and oriented at a
right angle to the basement membrane,
prominent nucleolus was common feature.
Interstitial cells of Leydig (ICL) were found in
the interstitial supporting tissue between the
seminiferous tubules. They were large rounded
or polygonal, some of them were small and
spindle (represent immature cells). Interstitial
cell of Leydig contained rounded vesicular
nuclei usually the cytoplasm was eosinophilicfig
(2).
Examination of H&E stained sections in
Se group (GII) showed no detectable variation
in comparison to the control group fig (3: A&B)
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 AL-AZHAR ASSIUT MEDICAL JOURNAL
showed the broad similarity between this group
and the control group.
Examination of Tr group (G III), H&E
stained sections showed apparently wider
interstitial spaces. Some tubules appeared to be
similar to those of the control, others showed
some disruption in their epithelial lining and
vacuoles, or empty spaces between their lining
spermatogenic cells fig (4). Higher
magnification showed that some of the tubules
appeared to be lined by single layer of
cells,however, other seminiferous tubules
appeared to be lined by many layers of
disrupted spermatogenic cells, most of these
cells were 1ry spermatocytes and few Sertoli
cells. These cells were interspaced by vacuoles
and acidophilic remnants. Spermatids and
sperms were hardly recognized within these
deteriorated tubules fig (5: A&B). Interstitial
tissue in between showed some disruption, some
Leydig cells appeared disintegrated or
vacuolated, in spite of that still harboring few
apparentlyhealthy interstitial cell of Leydigfig
(5: B).
Examination of Se and Tr group (GIV)
showed that the seminiferous tubules were
closely approximated to each other, few tubules
showed some small vacuoles between their
lining spermatogenic cells, most of the tubules
were broadly similar to the control fig (6).
Higher magnification, showed many
seminiferous tubules and interstitial tissues were
more or less similar to the control fig (7).
Electron microscopic results:
Examination of ultrathin testis sections of
the control group showed that the seminiferous
tubule appeared to be surrounded by basement
membrane (B.M.) and myoid cell. The B.M.
was regular and of uniform pattern, the myoid
cell was slender, smooth muscle like, it was
present outside the B.M. within the basal
lamina. The spermatogonia rested on the B.M.,
they were rounded with rounded euochromatic
nucleus and prominent nucleolus, many
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mitochondria could be seen scattered within the
cytoplasm fig(8).
The 1ry spermatocytes appeared to be
large cells with large euochromatic nuclei.
Spermatid appeared with large euchromatic
nucleus and its characteristic acrosomal vesicle ,
many sperms or fully formed spermatozoon
were also detected embedded within the Sertoli
cell, which appeared as a long dark cell with
irregular outline fig(9). Interstitial cell of Leydig
showed regular outline, containing euchromatic
indented nucleus fig (8).
Electron microscopic examination of Se
group (GII) showed no detectable variation than
the control group fig (10) support these
findings.
Exanimation of ultrathin sections of Tr
group (GIII) showed that the seminiferous
tubule lined by disrupted spermatogenic cells,
some of them contained dark condensed small
nuclei, some of these cells appeared ruptured, in
spite of that, the rarely seen spermatids were
recognized by the characteristic acrosomal
vesicle fig (11). The B.M. of the seminiferous
tubule was seen in some areas interrupted and
irregular, scattered collagen fibers were also
detected around it. Some of the interstitial cells
of Leydig appeared with irregular outline, some
otherswere ruptured, nuclei of such cells were
small, dark and condensed fig (12).
Examination of ultrathin sections of Se
and Tr (G IV) group showed generalized
amelioration of the previous findings. 1ry
spermatocytes, spermatids, and mature sperms
were clearly seen, more or less similar to the
control group. The B.M. of the seminiferous
tubule was continuous and regular.
However,some collagen fibers were still
recognized. The ICL was detected beside a
blood capillary that contained red blood
corpuscle (RBC), the ICL was of regular outline
and euchromatic nucleus with prominent
nucleolus fig (13).
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 Eman Badawi El Shal* and Mai M.H. Selim** VOL 13 , NO 1 , JANUREY 2015 SUPPL

Fig (1): Showing the seminiferous tubules in various planes of section, and interstitial tissue in
between.
Control, H&E x 100

Fig (2): Showing parts of 3 seminiferous tubuleslined by basalsperomatogonia with dark rounded
nuclei (1), 1ry spermatocytes with large pale nuclei (2), spermatids (3) and sperm (4),Sertoli cell
(5) and ICL (6) can be seen.
Control,H&E x 400

Fig (3):A, showing seminiferous tubules are cut in various planes of section. B, showing parts
seminiferous tubules lined by:spermatogonia (1), 1ry spermatocyte (2), spermatid (3), sperm (4),
ICL (arrow) can be seen.
(G II), H&E x100 (A), 400 (B)

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 AL-AZHAR ASSIUT MEDICAL JOURNAL VOL 13 , NO 1 , JANUREY 2015 SUPPL

Fig (4): showing seminiferous tubules, some of them contain many vacuoles (arrows), apparently
wide interstitial spaces.
(G III), H&E x100

Fig (5): showing different seminiferous tubules. (A): showing tubule lined by one layer of
unhealthy spermatogenic cells (1) interrupted by vacuoles (2) and acidophilic remnants (3). (B):
showing tubule lined by many layer of spermatogenic cells mainly 1ry spermatocytes (1), some
Sertoli cells (2), vacuoles(3), andacidophilic remnants (4). Healthy ICL (5), and vacuolated and
disintegrated ICL (6).
(G III), H&E x400

Fig (6): showing many seminiferous tubules are broadly similar to the control.
(G III), H&E x100

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 Eman Badawi El Shal* and Mai M.H. Selim** VOL 13 , NO 1 , JANUREY 2015 SUPPL

Fig (7): showing parts of different seminiferous tubules lined by ; 1- spermatogonia, 2-1ry
spermatocytes, 3-spermatids, 4-sperms, 5- Sertoli cells, ICL (6) are also noticed.
(G IV), H&E x 400

Fig (8): showing spermatogonium (S) with its large euchromatic nucleus (N) and prominent
nucleolus (n), many mitochondria (m), myoid cell (M), continuous B.M.(arrows).Inset: ICL
Control, TEM x 10000
Inset x 8000

Fig (9): showing part of seminiferous tubule lined by, 1ry spermatocytes (S1), Sertoli cell (Se),
sperms (S3). Inset: spermatid (S2) andacrosomal vesicle (AV).
Control, TEM x 3000
Inset x 5000
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 AL-AZHAR ASSIUT MEDICAL JOURNAL VOL 13 , NO 1 , JANUREY 2015 SUPPL

Fig (10): showing part of seminiferous tubule lined by 1ry spermatocytes (S1), Sertoli cell (Se),
sperms (S3). Lt inset: continuous B.M. (arrows) andmyoid cell (M). Rt inset: spermatid (S2)
andacrosomal vesicle (AV).
(G II), TEM x 3000
Lt. inset x 10000
Rt. inset x 5000

Fig (11): showing part of seminiferous tubule, disrupted spermatogenic cells, some of them
contained dark condensed nucleus (1), spermatid (S2) andacrosomal vesicle (AV). Inset: higher
magnification of part of this picture showing ruptured cell (arrows) and condensed dark nuclei
(2).
(G III), TEM x 3000
Inset x 5000

Fig (12): Lt side showing; ICL with irregular outline (arrow), ruptured cell (R) and condensed
nucleus (N). Rt side showing discontinuous B.M. (1), of irregular pattern (2), collagen fibers (C)
andmyoid cell (M).

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 Eman Badawi El Shal* and Mai M.H. Selim**
(G III),TEM Lt. side x 8000
Rt. side x 10000
Fig (13): showing part of seminiferous tubule, 1ry spermatocyte (S1), spermatids (S2), acrosomal
vesicle (AV) and sperm (S3). Lt inset: myoid cell (M), regular and continuous B.M. (arrows) and
collagen fibers (C). Rt inset: ICL and its nucleus (N), RBC within blood capillary.
(G IV), TEM x 3000
Lt. inset x 10000
Rt. inset x 8000
DISCUSSION
An increasingly alarming rate of tramadol
abuse has been revealed in the Egyptian
community in the last few years (Fawzi, 2011).
Generally, opioids were used as analgesic drugs
without standing the several side effects already
known. One of the side effects that rarely
considered was hypogonadism(Reddy,2010).
In the present study, in tramadol treated
group (G III) light microscopic examination
showed apparently wider interstitial spaces.
Some tubules showed some disruption in their
epithelial lining,some cells were replaced by
vacuoles or acidophilic remnants. Similar
results were detected by (Abdellatief et al.,
2014) they reported thattramadol affected testis
histomorphologically and induced variable
degrees of degeneration in the seminiferous
tubules. They showed shrinkage,
disorganization and vacuolization of
spermatogenic layers. Darkly stained nuclei
were sloughed into the tubular lumen. They
added that the mean values of seminiferous
tubules diameter and the mean germinal
epithelial height were significantly decreased.
This observation was in line with the
results of (Rashed et al., 2010) who accounted
the widening of peritubular tissue due to
withdrawal of gonadotrophic stimulation as
occurring after hypophysectomy. These
observations confirmed by (El-Gaafarawi2006)
VOL 13 , NO 1 , JANUREY 2015 SUPPL
who explained the cause of these changes by the
direct effect of tramadol on the hypothalamic–
pituitary axis leading to inhibition of secretion
of both FSH and LH. Both FSH and LH were
responsible for normal spermatogenesis in male
rats; therefore, decrease in peripheral LH
resulted in reduction of testosterone secretion,
which could be involved in the involution of the
seminiferous epithelium. An alternative way for
tramadol to suppress testosterone was to induce
nitric oxide (NO) (Ahmed and Kurkar, 2014).
Stevens and Lowe(1997) stated that
Sertoli cells are not affected by any of the
factors that injure the sensitive germinal
epithelium. This may explain why most of the
surviving cells in the current experiment were
1ry spermatocytes and Sertoli cells, Oxidative
stress in the testes is one of the major factors
that induce germ cell apoptosis (Rao and
Shaha, 2000; Yang et al, 2001). Light and
electron microscopic results in the present work
showed that the sperms and spermatids were
rarely seen, this may explained by (Bauché et
al., 1994) who said that the reactive oxygen
species (ROS) are known to be cytotoxic and
often cause tissue injury. Erkkila et al(2002)
stated that germ cells in late phases of
differentiation are most sensitive to death
inducing condition, also Eddy (1999)
mentioned that the spermatocytes arrested in
prophase meiosis and then eliminated at late
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 AL-AZHAR ASSIUT MEDICAL JOURNAL
pachyten phase by apoptosis resulting in
absence of spermatids.Maneesh et al. (2005) in
their experiment noticed histopathological
changes in the seminiferous tubules of rat testes
in the form of apoptosis of germ cells and
vacuoles of varying sizes, they explained that by
the excessive production ofROS due to alcohol
administration.
Furthermore, in the present work, electron
microscopic examination of the testes of group
(III) showed ruptured some of the
spermatogenic cells, these cells contained dark
small nuclei, the same changes were noticed
within the ICL this may be explained byNewton
etal.(2005) and Tamura et al.(2012) who said
that the free radicals facilitate the release of
lysosomal enzymes into cytosol with subsequent
oxidation of the protein components of the cell,
causing their fragmentation.In the present work
the effect of tramadol on the interstitial cells of
Leydig was explained by Alvarez andStorey
(1995) who suggested that tramadol could lead
to low testosterone secretion by these cells and
induced Lipid peroxidation can eventually result
in dysfunction and structural damage of cells.
The electron microscopic examination of
the same group showed that the B.M. was
irregular and discontinuous surrounded by
scattered collagen fibers. These findings were in
accordance with those of Ahmed and
Kurkar(2014) who reported that treatment with
tramadol causes a reduction in the activity of
scavenger antioxidant enzymes, leading to
overproduction of nitric oxide (NO) and
oxidative stress. Free radicals disturb B.M.
integrity through lipid peroxidation, this might
be the cause of discontinuous irregular pattern
of B.M.Berruti (2006) mentioned that the
thickness in the B.M. due to an increase
production of glycosaminoglycans and
proteoglycans and considered the collagen type
IV in the B.M. as a defense reaction against the
damaging activity of free radicals. Lombardo
et al (2005) said that certain stimulants may
induce myoid cells to produce more collagen
and extracellular matrix, which are responsible
for basal lamina thickness.
In the present study of Se and Tr group
(IV) showed improvement of many
seminiferous tubules and interstitial tissues
which were more or less similar to the
control.Fraga et al (1991); Kasahara et al
(2002) stated that the antioxidants protect germ
cell against oxidative DNA damage and play
important role in the spermatogenesis. The
improvement in the previous findings was
VOL 13 , NO 1 , JANUREY 2015 SUPPL
supported by Behne et al. (1988) who
suggested that Se was preferentially maintained
in the rodent testis. Se was essential for the
production of normal spermatozoa and thus
played a critical role in testis, sperm, and
reproduction (Flohé, 2007). In addition, Grotto
et al (2009) examined the possible
antigenotoxic effect of selenium in rats
chronically exposed to low levels of methyl
mercury,they detected that Se co-administration
showed a significant reduction in methyl
mercury-induced genotoxicity and reduced
DNA damage. Kara et al. (2007) concluded
that the mechanism of chemoprotection of
selenium may be related to its antioxidant
properties as well as its ability to interfere with
DNA repair pathways. It is thought that the
seminiferous epithelium and mature sperm also
require a particularly efficient protection against
oxidative stress which was provided by Se(Zini
and Schlegel, 1997).
Light and electron microscopic
examination of Se and Tr group (IV) showed
that the spermatid and sperms were clearly seen
and apparently healthy. The co-administrationof
Se and Trwas in line with Kaur and Bansal
(2005) who explained that Se had an
important role in protection and was the key
component of a protein present in the sperm,
which had a role in maintaining normal
fertility.Su et al (2008) added that Se could
strengthen antioxidant ability by enhancing
activities of antioxidant enzymes and by
increasing contents of the antioxidants.Se was
essential for the production of normal
spermatozoa and thus played a critical role in
testis, sperm, and reproduction (Flohé, 2007).
Ahmed and Kurkar (2014)and
Abdellatief et al.(2014)revealed that tramadol
treatment increased testicular level of nitric
oxide, and lipid peroxidation and decreased the
antioxidant enzymes activities, these may
facilitate the damage of testicular tissue, on the
other hand, EL-Gerbed(2013) mentioned that
the Se has the ability to counteract free radicals
and protect the structure and function of
proteins, DNA, and chromosomes against
oxidation injury. Even, Aslam et al.(2010)
stated that the Se is the most powerful trace
element in reducing storage and toxicity of
ROS. The previous authorsexplained why the
B.M. in (G IV) of the current experiment
wasseen continuous and of regular pattern
surrounded by some collagen fibers.The
collagen fibers which were present around the
B.M. in this group was explained by Stevens
134 | P a g e
 Eman Badawi El Shal* and Mai M.H. Selim**
and Lowe (1997) who described the B.M. of
the seminiferous tubule, and mentioned that it
was surrounded by C.T. basal lamina which
contained myoid cell and collagen fibers as a
usual findings in healthy tissue. This opinion
was supported by the healthy regular and
continuous B.M.
CONCLUSION
The use of tramadol, one of the centrally
acting analgesics, causes serious changes in the
testicular tissue that could threaten the
reproductive functions. Se ameliorated the
tramadol induced testicular toxicity to a great
extent.
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Antioxidant system in rat testicular
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10. Behne, D., Hilmert., H, Scheid, S.,
Gessner, H., Elger, W. (1988): Evidence for
specific selenium target tissues and new
biologically important selenoproteins.
BiochimBiophysActa. 966: 12–21.
11. Berruti, G. (2006): Signaling events
during male germ cell differentiation: update.
Front Biosci. 11:2144-56.
12. Brunton, L.L., Lazo, J.S., Parker, K.L.
(2006): Goodman and Gilman’s. “The
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13. Carson, F.L. and Hladik, C.
(2009):Histotechnology, A self instructional
text. 3rd ed. Chicago; American Society for
Clinical pathology. pp: 161-170.
14. Ceccarelli, I., De Pradova, A.M.,
Fiorenzani, P., Massafra, C., Aloisi, A.M.
(2006): Single opioid administration modifies
gonadal steroids in both the CNS and plasma
of male rats. Neuroscience. 140:929–937.
15. Drury, R.A. and Wallington,
E.A.(1980): “Histological techniques”. 5th ed.
Oxford NY, Toronto; Oxford University
press. pp: 520-535.
16. Eddy,E.M.(1999): Role of heat shock
protein HSP70-2 in spermatogenesis. Rev
Reprod. 4 :23-30.
17. El-Gaafarawi I.I. (2006): Biochemical
toxicity induced by tramadol administration
in male rats. Egypt J Hosp Med., 23:353–362.
18. El-Gerbed, M.S.A.
(2013):Histopathological and ultrastructural
effects of methyl parathion on rat testis and
protection by selenium. Journal of Applied
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S63.
19. Erkkila, K, Aito, H., Aalto,
K., Pentikäinen, V., Dunkel, L.
(2002):Lactate inhibits germ cell apoptosis in
the human testis. Mol Hum Reprod. 8(2):109-
117.
20. Fahmy, M.A., Hassan, N.H.A.,
Farghaly, A.A., et al (2008): Studies on the
genotoxic effect of beryllium chloride and the
possible protective role of selenium /vitamins
A, C and E. Mutat. Res., 652: 103-111.
21. Fawzi, M.M. (2011): Some medicolegal
aspects concerning tramadol abuse: the new
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22. Flohé, L. (2007): Selenium in
mammalian spermiogenesis. Biol Chem.,
388: 987–995.
23. Fraga, C.G., Motchnik, P.A.,
Shigenaga, M.K., et al (1991): Ascorbic acid
protects against endogenous oxidative DNA
damage in human sperm. Proc. Nalt. Acad.
Sci. USA. 88: 11003- 11006.
24. Grotto, D., Barcelos, G.R., Valentini,
J., Antunes, L.M., Angeli, J.P., Garcia,
S.C., Barbosa,J. F. (2009): Low levels of
methyl mercury induces DNA damage in rats:
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25. Hajibagheri, M.A.N. (1999): Electron
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Humanana press. Inc. Totowa and New
Jersey. (117): p- 3
26. Kara, H., Cevik, A., Konar, V.,
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Protective effects of antioxidants against
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27. Kasahara, E.1., Sato, E.F., Miyoshi, M.
et al. (2002): Role of oxidative stress in germ
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ethylhexyl)phthalate. Biochem. J. 365:849-
856.
28. Kaur, P., Bansal, M.P. (2005): Effect of
selenium-induced oxidative stress on the cell
kinetics in testis and reproductive ability of
male mice. Nutrition 21:351–357.
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histochemical methods, theory and practice.
3rd ed. ARNOLD. London, New York and
new Delhi. P: 31.
30. Koyuturk, M., yanardag, R., Bolkent,
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31. Lombardo, F.1., Sgrò, P., Salacone,
P., Gilio, B., Gandini, L. et al. (2005):
Androgens and fertility.J. Endocrinol.
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32. Maneesh, M., Jayalekshmi, H., Dutta,
S. et al. (2005): Role of oxidative stress in
ethanol induced germ cell apoptosis - An
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33. Messaoudi, M., Banni, L., Said, K., et
al (2010): Involvement of selenoprotein P
and GPx4 gene expression in cadmium-
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Chem. Biol. Interact. 188:94- 101.
34. Newton, A.P., Cadena, S.M., Rocha,
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triclosan (TRN) on energy-linked functions of
rat liver mitochondria.ToxicolLett. 160 :49-
59.
35. Nossaman, V.E., Ramadhyani, U.,
Kadowitz, P.J. et al (2010): Advances in
perioperative pain management: use of
medication with dual analgesic mechanisms,
tramadol and tapentadol. AnesthesiolClin.
28(4): 647- 677.
36. Paget, G.E. and J.M. Barnes,
(1964):Evalution of Drug Activites and
pharmacokinetics. Acadenic Press, Vol. I:
135-136.
37. Rashed, R.M., Mohamed, I.K., EL-
Alfy, S.H. (2010): Effects of two different
doses of melatonin on the spermatogenic cells
of rat testes: a light and electron microscopic
study. Egypt J Histol 33:819–835.
38. Reddy, R.G. (2010): Aung T, Karavitaki
N, Wass JAH. Opioid induced
hypogonadism. BMJ., 341:605–606.
39. RaoA.V.and Shaha, C.(2000): Role of
glutathione S-transferases in oxidative stress-
induced male germ cell apoptosis. Free
Radical Biol. Med. 29(10):1015-27.
40. Said, L., Banni, M., Kerkeni, A. et al
(2010):Influence of combined treatment with
zinc and selenium on cadmium induced
testicular pathophysiology in rat. Food Chem.
Toxicol. 48: 2759- 2765.
41. Stevens, A. and Lowe,T.
nd
(1997):”Human Histology”. 2 edition.
Mosby. London, Boston, New York, Tokyo
and Sydney. pp: 309-226.
42. Su, L., Wang, M., Yin, S.T., Wang,
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(2008): The interaction of selenium and
mercury in the accumulations and oxidative
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70 :483–489.
43. Swathy, S.S., Panicker, S., Indira,
M.(2006): Effect of exogenous selenium on
the testicular toxicity induced by ethanol in
rats. Indian J. PhysiolPharmacol. 50(3): 215-
224.
44. Tamura, I., Kanbara, Y., Saito,
M,, Horimoto, K., Satoh, M., et al.
(2012):Triclosan, an antibacterial agent,
increases intracellular Zn(2+) concentration
in rat thymocytes: its relation to oxidative
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 ‫**‪Eman Badawi El Shal* and Mai M.H. Selim‬‬ ‫‪VOL 13 , NO 1 , JANUREY 2015 SUPPL‬‬

‫‪45. Ursini, F., Heim, S., Kiess, M., et al‬‬ ‫‪protects against hydrogen peroxide-induced‬‬
‫‪(1999):‬‬ ‫‪Dual‬‬ ‫‪function‬‬ ‫‪of‬‬ ‫‪the‬‬ ‫‪apoptosis and inhibits JNK and caspase 3‬‬
‫‪selenoproteinPHGPx‬‬ ‫‪during‬‬ ‫‪sperm‬‬ ‫‪activation. The Journal of Biological‬‬
‫‪maturation. Science. 285: 1393-1396.‬‬ ‫‪Chemistry. 276: 19220-19230.‬‬
‫‪46. Yang, Y., Cheng, J., Singhal, S. S. et‬‬ ‫‪47. Zini, A., Schlegel, P.N. (1997):‬‬
‫‪al.(2001): Role of Glutathione S-Transferases‬‬ ‫‪Expression of glutathione peroxidases in the‬‬
‫‪in Protection against Lipid Peroxidation‬‬ ‫‪adult male rat reproductive tract. FertilSteril.‬‬
‫‪overexpression of Hgsta2-2 in K562 cells‬‬ ‫‪68: 689-695.‬‬

‫تأثير عالج انترايادول عهً خصً انجرر وانذور انىقائي انًحتًم نهسيهيُىو (دراسة بانًيكروسكىب انضىئً واإلنكتروًَ)‪.‬‬
‫إيى اٌ بذوي انشال* ويً يحًذ حسًُ سهيى**‬
‫قسى انرشش‪ٚ‬ح* ٔانٓسرٕنٕخٗ**– كه‪ٛ‬ح طة انثُاخ خايعح األصْش‬
‫ـــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــ ــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــ‬
‫انًقذية‪:‬‬
‫إٌ األنى يشكهح صح‪ٛ‬ح عظًٗ ٔاألدٔ‪ٚ‬ح اليخذسج ْٗ اإلخر‪ٛ‬اس األٔل نرخف‪ٛ‬ف حاالخ االنى انًرٕسطح ٔانشذ‪ٚ‬ذج ‪.‬‬
‫ٔسغى أٌ انرشايادٔل ْٕ يخذس نّ ذأث‪ٛ‬ش يشكض٘ إر أَّ ‪ٚ‬قهم يٍ خٕدج انسائم انًُٕٖ فٗ انًرعاط‪ ٍٛ‬نعقاس انرشايادٔل تشكم يضيٍ ‪,‬‬
‫ٔيٍ َاح‪ٛ‬ح أخشٖ ٔخذ أٌ انس‪ٛ‬ه‪ُٕٛ‬و انًضاد نألكسذج ضشٔسٖ الكرًال انًُٕ انطث‪ٛ‬عٗ نهخص‪ٛ‬ح‪.‬‬
‫انهذف يٍ انبحث‪:‬‬
‫دساسح انرغ‪ٛ‬شاخ فٗ ذشك‪ٛ‬ة خصٗ اندشر تالي‪ٛ‬كشٔسكٕب انضٕئٗ ٔاإلنكرشَٔٗ ٔانرٗ ذُرح تعذ ذعاطٗ عقاس انرشايادٔل ‪ٔ ,‬ذق‪ٛٛ‬ى دٔس‬
‫انس‪ٛ‬ه‪ُٕٛٛ‬و‬
‫انًىاد وانطرق‪:‬‬
‫ذى اسرخذاو أستعٌٕ خشرا قسًد إنٗ أستع يدًٕعاخ يرسأ‪ٚ‬ح انًدًٕعح األٔنٗ ْٗ انًدًٕعح انضاتطح ‪ ٔ ,‬انًدًٕعح انثاَ‪ٛ‬ح ذى‬ ‫ً‬
‫إعطاؤْا دٔاء س‪ٛ‬ه‪ُٕٛٛ‬و تدشعح ‪ 31.5‬و ‪ٚ‬كشٔخشاو ‪/‬خشر ‪ٕٚ /‬و‪ٔ ,‬انًدًٕعح انثانثح ذى إعطاؤْا انرشايادٔل تدشعح ‪7‬يدى‪ /‬خشر ‪ٕٚ /‬و‬
‫ٔخً‪ٛ‬ع اندشعاخ أر‪ٚ‬ثد فٗ انًاء انًقطش‪.‬‬
‫ٔانًدًٕعح انشاتعح ‪ :‬أعط‪ٛ‬د س‪ٛ‬هُ‪ٕٛ‬و يع عقاس انرشايادٔل تُفس اندشعاخ سانفح انزكش ‪ٔ ,‬قذ أعط‪ٛ‬د األدٔ‪ٚ‬ح عٍ طش‪ٚ‬ق انفى نًذج ‪8‬‬
‫أسات‪ٛ‬ع ثى خًعد ع‪َٙ‬اخ انخصٗ نهفحص تانًدٓش انضٕئ‪ٔ ٙ‬اإلنكرشَٔٗ‪.‬‬
‫انُتائج‪:‬‬
‫ٔخذ فٗاألَات‪ٛ‬ة انًُٕ‪ٚ‬ح اٌ انُس‪ٛ‬ح انطالئٗ تّ فدٕاخ ٔخال‪ٚ‬اِ يُضٔعح ٔيرفككح داخم األَات‪ٛ‬ة ٔعهٗ يسرٕٖ انً‪ٛ‬كشٔسكٕب‬
‫اإلنكرشَٔٗ فقذ ٔخذ أٌ انخال‪ٚ‬ا انًكَٕح نهح‪ٕٛ‬اَاخ انًُٕ‪ٚ‬ح تٓا فدٕاخ ٔإَٔ‪ٚ‬رٓا يُكًشح يع خهم ف‪ ٙ‬ذٕص‪ٚ‬ع انً‪ٛ‬رٕكَٕذس‪ٚ‬ا ٔأٌ خال‪ٚ‬ا ن‪ٛ‬ذج‬
‫تٓا خال‪ٚ‬ا دُْ‪ٛ‬ح ٔٔخذ أ‪ٚ‬ضا ً أٌ انغشاء انقاعذ٘ سً‪ٛ‬ك ٔحٕنّ أَسدح ن‪ٛ‬ف‪ٛ‬ح ٔتعذ اعطاء انس‪ٛ‬ه‪ُٕٛٛ‬و ٔخذ ذحسٍ ف‪ ٙ‬خً‪ٛ‬ع انرغ‪ٛ‬شاخ سانفح‬
‫انزكش ‪.‬‬
‫انخالصة‪:‬‬
‫ً‬
‫أٌ ذعاط‪ ٙ‬عقاس انرشايادٔل ذسثة فٗ إحذاز ذغ‪ٛ‬شاخ فٗ ذشك‪ٛ‬ة خصٗ اندشراٌ ‪ٔ ,‬تإعطاء انس‪ٛ‬هُ‪ٕٛ‬و يرضايُا يع عقاس انرشايادٔل أدٖ‬
‫إنٗ ذحسٍ ف‪ ٙ‬خً‪ٛ‬ع انرغ‪ٛ‬شاخ ‪.‬‬

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