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The Effect of Nandrolone Decanoate on the Testis
Weight and its Histological Structure in Mature
Male Albino Mice
Batool Ibrahim Hussain Shaima Ahmed Raheem Ikhlass M. A. Alsharifi
College of science for women / Univ. of Babylon
Abstract
This study aimed to identify the effects of different doses of Nandrolone decanoate (50 and 100 mg/kg )
subcutenously every 2 days for 30 days on testis weight and histological changes which include the number
of spermatogonia , spermatocyte , spermatide , lydige cells and the diameter of seminiferous tubules for
treatment group in comparing with control group which injected with normal saline .
The aim of this study to assess the effect of Anabolic androgen steriads AASs on spermatogonia ,
spermatocyte , spermatide , lydig cells , the diameter of seminiferous tubules of testis and the weight of
testis in adult male albino mice .
الخلاصة
50 و 100 ملغم/كغم) التي حقنت تحت ) Nandrolone decanoate هدفت الدراسة الحالية لمعرفه تأثير الجرع المختلفة من
الجلد كل يومين ولمدة 30 يوم على وزن الخصى ولمعرفه التغيرات ألنسجيه والتي تتمثل بعدد سليفات النطف ، الخلايا النطفيه، ارومات
النطف ، خلايا ليدك وأقطار النبيبات المنوية للحيوانات المعاملة مقارنه مع مجموعه السيطرة التي حقنت بالمحلول الملحي الفسلجي .
Introduction
Anabolic androgen steriods ( AASs ) are synthetic derivative of testosterone which
are pharmacologically important in the treatment of growth deficiency , some blood
disorders and osteosis ( Wood 2004 ; Feinberg etal.,1997) .
Several studies have reported that AASs have negative health consequence including
endocrine , hepatic , cardiovascular and behavioral disturbances (Iippi & Guidi, 1999;
Clark etal ., 1997) .
AASs compounds alter the function of the hypothalamic – pituitary– gonadal axis
and as a result affect the target reproductive tissues . some authors have reported that
AASs decreased density, motility and normal morphology of sperm (Holma, 1997;
Torres– Calleja et al., 2001) even low doses of AASs decrease sperm quality and quantity
(Dohle etal ., 2003). Mesbah et al . (2007) showed that administration of AASs caused
degerated change on some testicular structures . this adverse effect of AASs is due to its
inducing to circulating testosterone elevation to the range likely to be used in hormonal
male contraception (Daryl et al., 2004).
Material and Methods
Fifteen adult male albino mice (balb/c) weighting 18.4–22.8 g were randomized into
three groups : two groups treated subcutenously with 50 and 100 mg/kg body weight
nandrolone decanoate (ND). while control group was injected with 0.9 % normal saline ,
The animals were housed at 22-25 C° with 12 – h light / dark cycle .
After 30 days from injection ND the animals were sacrificed and their testes removed
and weight by sensitive digital balance . the organs to body weight ratio were then
calculated as mg/ 100 gm of body weight.
The testes were fixed by formaline 10% for histological sectioning ( Presnell &
Schreibman . 1997) . then the sections were examined by using compound microscope
Journal of Babylon University/Pure and Applied Sciences/ No.(5)/ Vol.(20): 2012
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and the measurements were cording by using ocular micrometer after calibrated with
stage micrometer . the number of spermatogonia , spermatocyte , for 12 seminiferous
tubules for each animal and 20 reading for lydig cells as well as 20 reading for the
diameters of seminiferous tubules for each animal also .
Results
In this study , the weight of testis of experimental animals was significantly ( P< 0.01)
lower than that in the control (Figure -1- ) The number of leydig cells and spermatide in
experimntal rats ( which injected with 50 and 100 mg / kg ) were significantly ( P<0.05 )
lower than those of control group ( fig 2,3 and 6 ), Our study did not exert any change in
the number of spermatogonia and spermatocyte in treated rats in comparing with control
( figer 5,6 ).
In the present study , a significant increase (P<0.01) in the diameters of seminiferous
tubules in experimental mice ( which injected with 50 and 100 mg/kg) was showed (fig 4)
, in addition to this findings our study showed that accumulation of odematous fluid in
the lumen of semininferous tubules of treated animal.
845.96
718.59
557.73
0
200
400
600
800
1000
Tesits weight
(mg)/100gm
body weight
concentration
100 50 control (mg/kg)
Fig.1
.
Figure (1):Testis weight ( mg / 100 gm ) of body weight in mail mice which treated in
different doses of nandrolone decanoate . L . S . D ( P? 0.01 ) = 132.84
Figure(2):Mean numbers of spermatid in male mice which treated in different doses
of nandrolone decanoate . L . S . D ( P? 0.05 ) = 22.95
183.19
40.88
60.08
0
40
80
120
160
200
mean number
of spermatid
concentration
(mg/kg)
100 50 control
Fig.2
.
1495
16.88
7.1 5.71
0
4
8
12
16
20
mean
numbers of
lydic cells
concentration
100 50 control (mg/kg)
Fig.3
.
Figure(3): Mean numbers of lydic cells in male mice which treated in different doses
of nandrolne decanoate . L . S . D ( P? 0.05 ) = 1.83
81.54
158.24
178.43
0
50
100
150
200
mean of
diameters of
seminferous
tubules
concentration
100 50 control (mg/kg)
Fig.4
.
Figure(4): Mean of diameters of seminferous tubules in male mice which treated in
different doses of nandroone decanoate . L . S . D ( P ? 0.01 ) = 7.56
51.72 48.91 46.33
0
20
40
60
80
100
mean of
numbers of
spermatogonia
concentration
100 50 control (mg/kg)
Fig.5
.
Figure(5): Mean numbers of spermatogonia in male mice which treated in different
doses of nandrolone decanoate .
Journal of Babylon University/Pure and Applied Sciences/ No.(5)/ Vol.(20): 2012
1496
56.82
47.97 54.3
0
20
40
60
80
100
mean
numbers of
spermatocyes
concentration
100 50 control (mg/kg)
Fig.6
.
Figure(6): Mean numbers of spermatocytes in male mice which treated in different
doses of nandrolone decanoate.
Picture (1):Cross – section ( 40 x ) for testis of treated mice with 50 mg / kgm shows :
degeneration of spermatogenic cells , inter space among seminiferous tubules , reduction
the number of spermatid , disappear of spermatozoa .A: spermatogonia . B:
spermatocyte . C: spermatid . D: spermatozoa E: diameter of seminiferous tubule.
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Picture(2):Cross – section (40 x ) for testis of treated mice with 100 mg / kgm shows :
degeneration of spermatogenic cells , reduction the number of spermatid ,
accumulation of odematous fluid inter tubules .A: spermatogonia .
B:spermatocyte . C: spermatid . D: spermatozoa E: diameter of
seminiferous tubule.
Picture(3):Cross – section (40 x) for testis of treated mice with 50mg/kgm shows :
degeneration of spermatogenic cells , disappear of spermatozoa increase the diameter of
seminiferous tubules , accumulation of odematous fluid inter the tubules.B: spermatocyte
. C: spermatid . D: spermatozoa E: diameter of seminiferous tubule.
Journal of Babylon University/Pure and Applied Sciences/ No.(5)/ Vol.(20): 2012
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Picture(4):Cross – section (10 x ) for testis of treated mice with 50 mg / kgm shows :
degeneration of spermatogenic cells , interspaces among seminiferous tubules .
E: diameter of seminiferous tubule
Picture (5):Cross – section (10 x ) for testis of treated mice with 100 mg / kgm shows :
interspace among seminferous tubules , degeneration of spermatogenic cells ,
accumulation of odematous fluid inside tubules.E: diameter of seminiferous tubule
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Picture(6):Cross-section(40x)for testis mice(control group).show:normal view for
semniferous tubules.A: spermatogonia . B: spermatocyte . C: spermatid . D: spermatozoa
E: diameter of seminiferous tubule.
Discussion
In this study , the weight of testis of experimental animals was significantly P< 0.01)
lower than that in the control (Figure -1-) . this result is in agreement with Mesbah et al. ,
2007.
It is clearly known that gonadotropin releasing hormones from pituitary gland ( FSH
and LH ) have growth promoting effects on testis development and the administration of
exogenous androgens suppresses the serum LH and FSH level in human and rats (Torres
et al ., 2001 ; O’sullivan et al., 2000) . the exogenous testosterone has negative feed –
back effects on hypothalamic – pituitary – gonadal axis (Ludwig , 1950 ).
Under normal condition , LH is regulated by GnRh which released by hypothalamous
. LH interact with receptors on the leydig cells to produce testosterone which is then
transported to the testis and accessory reproductive organs for regulation of growth and
maintenance of these tissue . following administration of exogenous AASs , the high
level of androgen causes a decrease in LH release from the pituitary gland , which inturn
results in suppression of endogenous testosterone ( Fembery et al ., 1997 ; Lumia et al .,
1994) consequently , for decreased level of testosterone , testicular atrophy occurs
(Ludwig , 1950 ) .
The number of leydig cells and spermatides in experimntal rats ( which injected with
50 and 100 mg / kg ) were significantly ( P<0.05 ) lower than those of control group ( fig
2,3 and 6 ) . this result was consistent with others ( Mesbah etal ., 2008 and Grokett etal .,
1992 ) which they found sever depletion of leydig cells following treatment by AASs .
Journal of Babylon University/Pure and Applied Sciences/ No.(5)/ Vol.(20): 2012
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leydig cells are known to have receptors for LH that stimulates these cells to produce
testosterone ( Johnson et al ., 1972 ) .
Both LH and testosterone are responsible for normal spermatogenesis in male rats
(Zirkin , 1998) . there fore , depletion of LH receptors and decrease in peripheral LH by
exogenous testosterone administration result in reduction of testosterone secretion which
is necessary for formation of spermatide (Ichihara et al ., 2001) .
Our study did not exert any change in the number of spermatogonia and spermatocyte
in treated rats in comparing with control (figs. 5,6 ) , this may be due to AASs affects on
testosterone level which is not necessary for production these cells
In the present study, a significant increase (P<0.01) in the diameters of seminiferous
tubules in experimental mice (which injected with 50 and 100 mg/kg ) was showed (fig.4)
this increase may be due to decrease thickness of basement membrane in the
semininferous tubules of treated animals .
This is in accordance with other reports presented negative correlation between
basement membrance thickness and seminiferous tubules diameter (Aydes et al., 1998 ;
Santero et al ., 1999 ) , or may be due to decrease in testosterone concentration and only
the optimal levels of androgen can keep the normal structure of semininferous tubule
(Doust et al ., 2007 ) .
in addition to this findings our study showed that accumulation of odematous fluid
in the lumen of semininferous tubules of treated animal this due to increased permeability
of blood vessels lead to odema between cells and release large amount of plasma from
capillaries to the damge areas flowed by coagulation of tissue fluid and migration of
white blood cells ( Neutrophil and Monocyte ) to the in flammutory region by chemotaxic
agents that produced by damaged inflammatory tissues (Al–Saadi , 1992) .
In conclusion , a administration of Nandrolone decanoate exerts a clear effect on
testicular structure including degenerated changes of germ cells and leydig cells . Our
results suggest study the fine structure of the testis which affected with Nandrolone
decanoate by using electronic microscope .
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