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EVALUATION THE ROLE OF SUPPLEMENTATION OF VIT. D
ON SOME PHYSIOLOGICAL PARAMETERS, SEMEN
QUALITY IN HYPOVITAMINOSIS D MALE RABBITS
INDUCED BY FUROSEMIDE
Ayat R. Sabeeh , Muna H. AL-Saeed
*Department of Physiology, Pharmacology and Biochemistry, College of Veterinary
Medicine, University of Basrah,Basrah,Iraq
(Received 8 August 2020 ,Accepted 28 September 2020).
Keywords: Vit. D., Parathyroid hormone, Semen quality.
Corresponding Author: [email protected]
ABSTRACT
This study was planned to assess the improvement effect of supplementation
Vit.D on adult male rabbits with hypovitaminosis D was induced by put the male
rabbits in dark room un exposure to sunlight, eaten restricted food for experiment
period and administration of furosemide drug (0.5 mg/kg.B.W.). Vitamin D treated
rabbit group was given Vitamin D. two doses (high and low doses).Thirty adult male
rabbits of (1500-2000 gm) body weight were used in this study and animals were
divided into five main groups, each of 6 rabbits. Control group treated with normal
saline, second group the male rabbits put in dark room un exposure to sunlight, eaten
restricted food and treated with furosemid 0.5mg/kg B.W. I.M for 16 days, third
group the male rabbits put in dark room un exposure to sunlight, eaten restricted food
and treated with 0.5 mg/Kg B.W. of furosemid I.M for 16 days +200000IU/2ml of
Vit.D in every week for 1month, fourth group the male rabbits put in dark room un
exposure to sunlight, eaten restricted food and treated with 0.5mg/Kg B.W.of
furosemide I.M for 16 days+100000IU/1ml of Vit.D in every week for 1month, fifth
group put the male rabbits in dark room un exposure to sunlight, eaten restricted food
and treated with both 0.5 mg/kg B.W. of furosemid I.M for 16 days + exposure to
sunlight for 5 hours in day until 1 month. The taken results exposed that
hypovitaminosis D is associated by a significant (P≤0.05) decrease in body weight
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and body weight gain as compared with control (-ve) group. Also the results
indicate a significant (P≤0.05) decrease in serum Vitamin D, Calcium, total protein
and testosterone concentrations compared with control(-ve) and another treated
groups. A significant(P≤0.05) increase in serum concentrations of urea, creatinine,
uric acid, Phosphate, Parathyroid hormone (PTH), Follicle stimulating hormone(FSH)
and Luetinzing hormone(LH) have been shown in serum of hypovitaminosis D
groups compared to control(-ve) group. Semen analysis showed a significant decrease
(P≤0.05) of epididymal sperm concentration, sperm motility, and viability and a
significant increase (P≤0.05) of sperm abnormalities were recorded in hpovitaminosis
D groups compared with control(-ve) and another treated group. Also, histological
examination on the parathyroid gland, kidneys and testis showed histological changes
of hypovitaminosis D male rabbits treated with furosemide while when treated Vit.D
groups and group exposure to sunlight showed alimentative in list previous
parameters also histological examination.
INTRODUCTION
Vit.D is a lipophil secosteroid that are naturally known for Ca-P and bone
metabolism and pivotal target organs are intestine, skeletal system, renal and
parathyroid glands (1). Vit.D lifes in two forms; ergocalciferol (vitamin D2) and
cholecalciferol (vitamin D3) (2). These two forms differ in one double-binding and
one methyl group. While Vit.D2 is found in vegetable sources like mushrooms, Vit.D3
can be originate of animal sources like fat fish, cod oil of liver, yolk of egg and
fortified food like dairy products (2) or by endogenous skin synthesis that needs
exposure of sunbeams. Vit.D supplements might contain both forms, although in
Norway, vitamin D3 is almost exclusively used in over-the-counter Vit.D
supplements. Despite of its creating formation, pre-vitamin D suffers hydroxylation to
25-hydroxyvitamin D (25(OH)D) by 25-hydroxylase, a cytochromeP450-dependent
enzyme.
In 21st century, Vit. D deficit has happened to a global trouble changing 1
billion people (3). Severe Vit.D insufficiency reasons osteomalacia or rickets,
osteoporosis and bone breaks as of diminished calcium absorption (4). At the side of
its found character in skeletal strength, little Vit.D grade is conferred as a danger
aspect in next of kin to numerous non-skeletal health results for example CVD (5),
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fatness (6), diabetes (7), asthma (8), manifold sclerosis (MS) (9), cancer diseases (10)
and overall mortality (11, 12).
Diuretics are broadly applied in cardiovascular disease. Furosemide (Furo)
influence on parathyroid hormone circulating (PTH) and whether such deeds are
concluded by the results of these composites on Ca secretion (34). The aimed of this
work was studying the physiologic effect of Vit.D on parathyroid gland activity and
fertility of male through comparing the effect of deficiency and supplementation.
MATERIAL AND METHODS
This study was performed on thirty male rabbits weighing (1500-2000gm).
The rabbits were put in the animal house of College of Veterinary Medicine /
University of Basrah.
Twenty four rabbits were induction hypovitaminosis D. This rabbits put in
plastic cages in dark room un exposure to sunlight with restricted food for
hypovitaminosis D and drinking ad libitum under 20-25°C controlled temperature
condition.
Rabbits were divided into five main groups, each of 6 rabbits. Control(-ve) group
treated with normal saline, second group control positive (+ve) male rabbits put in
dark room un exposure to sunlight, eaten restricted food for experiment period and
treated with furosemid 0.5mg/kg B.W. I.M for 16 days(induction hypovitaminosis D),
and remain until end experiment, third group male rabbits put in dark room un
exposure to sunlight, eaten restricted food and treated with 0.5 mg/Kg B.W. of
furosemid I.M for 16 days +200000IU/2ml of Vit.D every week for 1moth, fourth
group male rabbits put in dark room un exposure to sunlight, eaten restricted food
and treated with 0.5mg/Kg B.W.of furosemide I.M for 16 days+100000IU/1ml of
Vit.D every week for 1moth, fifth group male rabbits put in dark room un exposure to
sunlight, eaten restricted food and treated with both 0.5 mg/kg B.W. of furosemid
I.M for 16 days + exposure to sunlight for 5 hours in day until 1 month.
At the end of experiment, rabbits were collection blood from heart puncture for
estimation studied parameters after that sacrificed, testis and epididymis were taken
out for semen analysis. Epididymal sperm were counted as illustrated by (13). The
individual and massive epididymal sperm motility was then approximation as
described by (14). Abnormal and dead sperms percentage were then records in the
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stained slide by eosin- nigrosin stain as illustrated by (13).Statistical Analysis was
carried out by one-way covariance (ANOVA) test. By SPSS program V. 21.(15).
RESULT
1-Effect of Supplementation of Vit.D on Body Weight and Body Weight gain in
Hypovitaminosis D Male Rabbits.
The current study in Table (1) discovered a significant increase (P≤0.05) in
final body weight in group drench 100000IU/1ml of Vit. D than control(+ve) while
the results illustrated no significant changes (P˃0.05) final body weight of group
drench 200000IU/2ml of Vit.D. and exposure to sunlight than control(-ve) and
control(+ve).
The results of body weight gain that demonstrated a significant (P≤0.05) high in
group drench 100000IU/1ml of Vit. D than another groups but no significant changes
(P˃0.05) of body weight gain in male rabbits treated with 100000IU/1ml of Vit. D
than control (-ve).
Table(1)Effect of supplementation of Vit.D on Body Weight and Body Weight gain
in Hypovitaminosis D Male Rabbits. Mean±SD N=6
Different letters indicate differences among groups at (P≤0.05) level vs. control. NS=Non significant.
Parameters
Treatments
Intial Body
Weight
(g)
Final Body
Weight
(g)
Body Weight
Gain
(g)
Control(-ve)
(Normal saline 0.9%NaCl)
١٦٣٠.00±١١٣.66
a
1816.70±132.91
ab
186.67±67.72
a
Control(+ve)
Hypovitaminosis D
By Furosemide
1686.70±146.24
a
1753.30±146.65
b
66.66±28.04
c
Hypovitaminosis D+
200000IU/2ml of Vit.D
1726.70±121.92
a
1848.30±121.55
ab
121.67±25.62
b
Hypovitaminosis D+
100000IU/1ml of Vit.D
1700.00±187.08
a
1948.30±179.49
a
198.33±31.88
a
Hypovitaminosis D+
Exposure of Sunlight
1683.30±194.07
a
1793.20±177.40
ab
109.83±50.32
bc
LSD NS 195.00 55.00
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2-Effect of Supplementation of Vit.D on Urea, Creatinin, Uric acid and Total
protein in Hypovitaminosis D Male Rabbits.
The results in Table (2) indicate a significant (P≤0.05) raise in serum urea,
creatinine, uric acid levels of hypovitaminosis male rabbits treated with furosemide
(control (+ve)) group as contrast with the control (-ve) group and another treated
groups. While no significant (P˃0.05) differences in the group serum urea, creatinine,
uric acid concentrations of male rabbits treated with 200000IU/2ml of Vit.D,
100000IU/1ml of Vit.D and group exposure to sunlight.On other hand, a significant
(P≤0.05) decrease in serum total protein concentration of hypovitaminosis male
rabbits treated with furosemide (control (+ve)) as compared with the control (-ve)
and treated with200000IU/2ml of Vit.D. But no significant (P˃0.05) differences in
serum total protein concentration of hypovitaminosis male rabbits treated with
furosemide (control (+ve)) as contrast with of male rabbits treated with 100000IU/1ml
of Vit.D and exposure to sunlight.
Table (2): Effect of Supplementation of Vit.D on Urea, Creatinin, Uric acid and Total
protein in Hypovitaminosis D Male Rabbits.(Mean±SD) (N=6)
Different letters represent differences among groups at (P≤0.05) level vs. control
Parameters
Treatments
Urea
mg/dl
Creatinine
mg/dl
Uric acid
mg/dl
Total
Protein
g/dl
Control(-ve)
(Normal saline
0.9%NaCl)
35±1.41
c
0.29±٠ .03
b
5.00±2.00
b
6.00±0.83
a
Control(+ve)
Hypovitaminosis D
By Furosemide
52.16±8.61
a
5.66±0.87
a
8.3±2.94
a
4.00±0.89
bc
Hypovitaminosis D+
200000IU/2ml of Vit.D
36.50±2.88
bc
0.37 ±٠ .051
b
4.16±1.72
b
6.41±0.73
a
Hypovitaminosis D+
100000IU/1ml of Vit.D
42.66±4.58
b
0.51± 0.055
b
4.66±0.81
b
5.25±0.52
b
Hypovitaminosis D+
Exposure of Sunlight
39.50 ±7.39
c
0.59±0.077
b
5.33±1.86
b
4.50±1.04
b
LSD 7.66 5.06 3 1.16
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3-Effect of Supplementation of Vit.D on Vit.D Ca, P and PTH Concentrations in
Hypovitaminosis D Male Rabbits.
The results of concentrations of Vit.D and Ca in male rabbits treated with
furosemide alone are represented in the Table(3). The results indicate a significant
(P ≤ 0.05) decrease progressively in serum Vit.D and Ca concentrations of male
rabbits treated with furosemide as control (+ve) than control (-ve) and another treated
groups. While treated with furosemide led to significant (P≤ 0.05) elevate in serum
PTH and P than control (-ve) and another treated.
Table(3):Effect of Supplementation of Vit.D on Vit.D, PTH, Ca and P Concentrations
in Hypovitaminosis D Male Rabbits. (Mean±SD) (N=6).
N=number of animals., different letters refer to differences among groups at (P≤0.05) level vs. control
Parameters
Treatments
Vit. D
ng/ml
PTH
Pg/ml
Ca
mg/ml
P
mg/ml
Control(-ve)
(Normal saline 0.9%NaCl)
48.93±6.40 a 5.68±0.11 b 13.70±1.35 a 5.73±0.56 b
Control(+ve)
Hypovitaminosis D
By Furosemide
17.83±1.72b 15.16± 2.22 a 9.45±1.44 b 10.98±0.03
a
Hypovitaminosis D+
200000IU/2ml of Vit.D
60.46±2.78 a 6.93± 0.94 b 13.28±2.04a 5.30±0.13 b
Hypovitaminosis D+
100000IU/1ml of Vit.D
41.16±4.40 a 6.66±0.57b 13.28±1.98a 5.76±0.88 b
Hypovitaminosis D+
Exposure of Sunlight
33.01±6.38ab 7.16±0.81 b 12.25±2.31a 6.63±1.67 b
LSD 19.30 8 2.79 4.35
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4-Effect of Supplementation of Vit.D on FSH, LH and Testosterone in
Hypovitaminosis D Male Rabbits.
The data in the table (4) showed a significant (P≤0.05) increase in the serum
FSH concentration of hypovitaminosis D male rabbits (control(+ve)) than the control
group(-ve) and another treated. However there were no significant(P˃0.05) variations
in serum FSH level among groups such as group treated 200000IU/2 ml of Vit. D,
treated with 100000IU/1ml of Vit. D, group exposure to sunlight and control(-ve).
The results also showed prominent role of furosemide induction
hypovitaminosis D in causing a significant (P≤0.05) increase in serum LH
concentration as compared to the control group(-ve) and another treated except group
drench 100000IU/1ml of Vit.D(low dose). While, significant (P≤0.05) decline in LH
concentration were observed in control(-ve) group than another treated groups.
Moreover, a significant (P≤0.05) decrease in serum T concentration was
recorded in control(+ve) hypovitaminosis D as compared to the control(-ve) and group
treated 200000IU/2ml of Vit. D(high dose).While no significant (P˃0.05) change in
serum T concentration observed as compared to control (+ve) and group treated
100000IU/1ml of Vit.D and group exposure to sun light.
Table (4): Effect of Supplementation of Vit.D on FSH, LH and Testosterone in
Hypovitaminosis D Male Rabbits. (Mean±SD) (N=6)
Parameters
Treatment
FSH
(mlU/ml)
LH
(mlU/ml)
Testosterone(T)
( ng/ml)
Control(-ve)
(Normal saline
0.9%NaCl)
٥.٥٠±٢.٤٢
b
3.30±1.35
d
9.80±1.30
a
Control(+ve)
Hypovitaminosis D
By Furosemide
١١.٨٣±1.94
a
11.08±1.42
a
5.57 ±1.95
b
Hypovitaminosis D+
200000IU/2ml of Vit.D
6.34 ±1.75
b
6.05±1.41
b
8.44±2.06
a
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Different letters indicate differences among groups at (P≤0.05) level vs. control,
5-Effect of Supplementation of Vit.D on Physical Properties of Semen Analysis in
Hypovitaminosis D Male Rabbits.
The current study in Table (4-11) illstrutled a significant raise (P≤0.05) in
mass activities, concentrations of sperm, total sperm and live sperm of male treated
with 200000IU/2mland 100000IU/1ml of Vit.D as than to control (-ve) and another
treated groups. However significant decrease (P≤0.05) were noted in semen volume,
mass activities, concentration of sperm, total sperm and live sperm of male treated
with than to control (-ve) and another treated groups.
There are a significant decrease (P≤0.05) in sperm abnormalities of male treated
with 200000IU/2ml of Vit.D than to control (-ve) and another treated . However,
significant increase (P≤0.05) were observed in sperm abnormalities of
hypovitaminosis D male rabbits than control(-ve)group and another treated groups.
Hypovitaminosis D+
100000IU/1ml of Vit.D
5.30±٢.٠٥٠
b
3.45±1.02
a
6.00±1.78
b
Hypovitaminosis D+
Exposure of Sunlight
٦.٠٨±1.93
b
٥.٥٣ ±1.93
c
6.50±1.64
b
LSD ٥.48 2.07 2.36
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Table(5): Effect of Supplementation of Vit.D on Physical Properties of Semen
Analysis in Hypovitaminosis D Male Rabbits. (Mean ±SD) (N=6)
N=number of animals., different letters refer differences among groups at P≤0.05 vs.
Hypovita D
minosis
D+
Exposure
of
Sunlight
Hypovitami
nosis D+
100000IU/1
ml of Vit.D
Hypovitami
nosis D+
200000IU/2
ml of Vit.D
Control(+ve)
Hypovitami
nosis D
By
Furosemide
Control(-ve)
(Normal
saline
0.9%NaCl)
Groups
Parameters
Semen colour Creamy Creamy Creamy Creamy Creamy -
Mass ++++ + ++++ +++ ++ -
activities
66.66±6.2 6.83
5
c
69.66±3.44
c
76.50±2.25
b
47.00± 5.05
d
86.33 ±3.98
a
Sperm
motility%
75.00±0.1 16
3
d
89.16 ± 0. 68
c
96.16±0.28
b
55.50±0.14
e
118.33 ± 4.50
a
Sperm
concentration
(×106/ml)
82.66:17.3 7.83
4 ±3.88
b
90.50:9.50
±4.03
a
91.50:8.50
±6.77
a
73.50:6.50
±2.66
c
92.50:7.50
±1.87
a
Live-dead
sperm ratio
15.8
0
41.20±6.3
7
b
35.83±5.63
c
16.66±3.88
d
57.00±14.56
a
15.33±1.03
d
Sperm
abnormalities
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6-Sperm Morphology
Fig.(1):-Sperm of rabbits (control(-ve)). Showing normal
sperm(N), Live(L), Stain with(N&E) 100X.
L
N
Fig.(3):-Sperm of rabbits treated with furosemide and 200000
IU/2ml (high dose) of Vit.D. Showing normal sperm(N),
Live(L), Stain with(N&E) 400X.
N
L
Fig.(2):- Sperm of rabbits (control(+ve)) treated with
furosemide. Showing abnormal shape sperm(A), most sperms
dead (D), some sperm tailless, many sperm with coiled tail (C),
sperm presented with Double head(H), Stain with(N&E) 100X.
A
D
C
H
Fig.(4):-Sperm of rabbits treated with furosemide and 100000
IU/1ml (low dose) 0f Vit.D. Showing normal sperm(N), Live(L),
Stain with(N&E) 400X.
L N
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7-Histological Examination:
Parathyroid Gland: Section of the parathyroid gland of male rabbits (control group) (-
ve).normal structure of parathyroid gland. There are two types of cells within the
parathyroid gland, the chief cells and the oxyphil cells (dark and light cells) as shown
in figure (6). While section of the parathyroid gland of male rabbits (control
group)(+ve) treated with furosemide induced hypovitaminosis D shows hyperactivity
with normal limits of structure as shown in figure (7). Also another groups treated
with Vit.D 200000 IU/2ml, 100000 IU/1ml and exposure to sunlight showed normal
parathyroid gland structure as shown as in figure (8),(9) and (10).
Kidneys: As shown in figures (11) kidneys of male rabbits of control (-ve) group
showed normal architecture of kidneys, normal glomeruli, normal renal cortical tubules
and normal epithelial cells lining of the renal tubules. While kidneys of male rabbits
treated with furosemide (control +ve) revealed histological changes as shown in
figure(12). The changes included infiltrations of inflammatory cells, congestion of renal
vascular, glomerular congestion. But kidneys of male rabbits treated with 200000
IU/2ml of Vit.D (high dose) showed normal architecture as shown in figures (13). In
adding, the renal of rabbits treated with 100000 IU/1ml of Vit.D(low dose) revealed
hemorrhage in the renal pelvis as shown in figures (14). Also the kidneys of rabbits
exposure to sunlight revealed interstitial medullary hemorrhage in figure(15).
Fig.(5):-Sperm of rabbits treated withfurosemide and exposure to sunlight.
Showing normal sperm(N), Live(L) and some sperm dead, Stain
with(N&E)400X.
L
D N
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Testes: Section of testis of control (-ve)group rabbits showing normal structural
design of seminiferous tubules as shown in figure(16).
While section of testis of rabbits treated with furosemide alone for induced
hypovitaminosis. Showing vacuolation of spermatognia and suppression of
spermatogenesis, widening of inter seminiferous tubules, decrease in the number of
interstitial leydig cells as shown in figure (17). But, section of testis of male rabbits
treated with Vit.D. 200000 IU/2ml (high dose). Showing normal of spermatogenesis
in seminiferous tubuled as shown in figure(18).
Also section of testis of rabbits treated with Vit.D. 100000 IU/1ml (low dose)
showing normal seminiferous tubules and suppression of spermatogenesis as shown
in figure(19). Also section of testis of male rabbits group exposure to sunlight,
showing normal seminiferous tubules, marked suppression of spermatogenesis as
shown in figure(20).
Fig (7): Section of Parathyroid gland of male rabbits control (+ve)
groups shows appears hyperactivity (greenarrow).H&EA)125X B)
500X
B A
Fig.(6):Section of Parathyroid gland of male rabbits control (-ve)
groups reveal normal parathyroid gland normal structure of chief
and oxyphil cells (dark(D) and light(L) cells. Stain (H&E) 400X.
D
L
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Fig.(8):Section of Parathyroid gland of male rabbits with
200000IU/2ml of Vit. D.(high dose), reveal normal parathyroid
(green arrow). A) 500X H&E
Fig.(10):Section of Parathyroid gland of male rabbits exposure to
sunlight shows appears within normal contain chief and oxyphil cells
(dark(D) and light(L) cells) hyperactivity limits (green
arrow).H&EA)125X B)500X
D
L
Fig(11):Section of kidney of male rabbits control (-ve). Showing
normal glomeruli (G) and normal renal cortical tubules (CT), stain
(H&E).A)125X B)500X
G
CT
Fig (9):Section of Parathyroid gland of male rabbits treated
with 100000 IU/1ml of Vit. D.(low dose) shows appears
within normal limits (green arrow). H&E A)125X
B)500X
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Fig (14) Section of kidney of male rabbits exposure to sun light
treated group, reveal interstitial medullary hemorrhage (green arrow).
A) 125 X B) 500 X H&E
B A
Fig. (15): Section of kidney of male rabbits treated with 0.1ml of
Vit.D (low does), reveal hemorrhage in the renal pelvis (green
arrow). A) 125 X H&E
Fig. (12): Section of kidney of male rabbits treated with 0.2ml of
Vit.D (high does), reveal normal collecting tubules in the medulla
(blue arrow), normal renal pelvis (green arrow).125 X H&E
Fig.(13): Section of kidney of control(+ve) group shows
glomerular congestion (green arrow), renal vascular
congestion (black arrow). A)50 XB) 500X
B A
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Fig: (19) Section of testis of male rabbits treated with 0.1ml of Vit.D (low
does), reveal normal seminiferous tubules (green arrow), Marked
suppression of spermatozoa (yellow arrow). A)125 X ,B)500 H&E
Fig.(16):Section of testes of male rabbits of control (-ve) group
shows normal seminiferous tubules (green arrow), normal
spermatozoa (black arrow). H&E A) 125X B)500X
Fig (17) Section of testes ofmale rabbits of control (+ve) group shows
marked vascular congestion (green arrow), marked atrophy of some
seminiferous tubules (black arrow), marked suppression of
spermatogenesis (blue arrow). H&E A)50X B)500X.
B A
Fig.(18): Section of testis of male rabbits treated with 0.2ml of
Vit.D (high does) reveal normal seminiferous tubules (green
arrow), normal spermatozoa (black arrow). H&E 100X.
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DISCUSSION
Vitamin D is the main steroid hormone which vitally essential for good bone- and
overall health the full time life in mishmash with Ca, P and PTH. It helps in the
absorption of Ca as of the intestine and reabsorption as of tubules of kidneys and in
combination with Ca mobilization as of bones by PTH(16). This delicate balance of
Vit.D, Ca, P and PTH is disturbed when impaired kidney progresses and this leads to
highly risk of not only bone disease but also cardiovascular disease(17).
Vit.D insufficiency raises the danger of expanding rickets, osteomalacia,
osteoporosis moreover add to the hazard of various non-skeletal adverse health
outcomes including CVD, autoimmune diseases and overall mortality. In animals, Vit.
D is mainly created in the skin following environmental exposure and only a little
quantity is get during the diet. The lifestyle, humans have a mixture of Vit.D2 and D3
available as of ambient ultraviolet (UV) rays, usual dietary intake of Vit. D3-rich
foods (yolks of egg and oil of fish), fortified foods which usually have vitamin D2
fortification(18). The extended shortage of 1,25 dihydroxy Vit.D together with
parathyroid abnormality leads to renal osteo-dystrophy (ROD) and CRDmineral bone
disorder (CKD-MBD), which can be evidenced by any one or mixture of
abnormalities of Ca, P, PTH, and Vit.D metabolism, leading to elevated in PTH2,
bone disease, vascular or soft tissue calcification.(19)and (20).
Fig.(20): Section of testes of male rabbits group exposure to sun light shows normal reveal
normal seminiferous tubules (black arrow), suppression of spermatozoa (green arrow). A)125 X,
B) 500X H&E
B A
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Parathyroid hormone is essential in Ca and P hemostasis and also plays a role in
the conversion of Vit.D to its active form (1, 25 dihydroxycholecalciferol) in the
renal. A reduce in extracellular Ca concentrations or an increase in P concentrations
guides to a PTH discharge as of the parathyroid gland, which in turn elevates renal
reabsorption of Ca, renal activation of Vit.D, urinary P secretion, and bone resorption.
In turn, Vit.D promotes intestinal absorption and renal reabsorption of Ca and
P(21),(22).
Diuretics are generally used with hypertension. We required to explore, in
hypertension, the influence of furosemide (Furo) on circulating parathyroid hormone
(PTH) and whether such actions are established by the results of these complexes on
Ca excretion.Furosemide elevated the hazard of HPT2. This effect was independent of
eGFR or Ca excretion. The use of LD, present preferred in advanced stages, should be
reappraised.
The present study revealed in Table (1) a decrease in body weight and body
weight gain in control (+ve) group along the experimental period. The decrease in
the body weight and body weight gain may be attributed to the postulated that
furosemide-induced reductions in body weight suggested reductions in body fluid
volume (23). Furosemide causes rapid and substantial urine loss, a 2% to 4% loss in
body weight, and, only 30 minutes after administration, a 13% reduction in plasma
volume(24). This profound increase in urination after furosemide dealing with in
horses, combined with its ability to cause increased excretion of Na, Ca, K and Cl, as
well as evidence in animals that furosemide dealing wirth may contribute to raised
danger of fracture.
The taken of furosemide caused serum Vit. D and Ca concentrations were
significantly reduced. This may reflect the activities of parathyroid hormone , which
may impair Ca regulation effects on thyroid gland and these changes may be
connected with effects on parathyroid gland therefore may cause disturbance in serum
concentration of Ca and P. The affect of serum Ca concentration in hypovitaminosis
rabbits reflect an adverse image on bone mineralization and this reveal the importance
role of thyroid hormones in regulation of growth and bone metabolism (25).
The obtained results revealed that a significant increase of creatinine and urea
concentrations in the serum of male rabbits treated with furosemide Control(+ve).
These results are attributed to deleterious effect of furosemide on kidney functions
and these results is supported by histological findings of kidney.
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Hypovitaminosis leads to adverse effects in various tissues. The present crosssectional
study was performed to determine whether furosemide has deleterious
effects on renal function. furosemide-induced kidney impairments in male rabbits.
In hypovitaminosis D directing to gout has been reported {26),(27). The raise in uric
acid in hypovitaminosis state may result from either enlarged production due to
myopathy associated with hypovitaminosis D or due to reduced renal clearance of uric
acid(28). Though the observed decrease in renal function, renal dysfunction induced
hypovitaminosis D may lead to adverse clinical. In this study, the histological changes
of the kidney especially the basement membrane of renal cortical tubules thickening
have been confirmed and agreed with hypovitaminosis D in animals. The reason is
low concentrations of Vit.D in active form, which is responsible for absorption and
reabsorption of Ca, due to the kidney losing their function, an enzyme is not
produced, 1-α hydroxylase responsible for convertingVit.D from formula in active 25-
hydroxycholecalciferol to active 1,25-dihydroxycholecalciferol(29),(30).
The obtained results revealed benefit effect of Vit.D supplementation plus
furosemide and exposure to sun light on serum concentrations of Vit.D, Ca and P in
male rabbits. Serum Vit.D was arrived to normal range in male rabbits after Vit.D
supplementation and exposure to sunlight. Also Ca and P concentrations get better
compared to treated with furosemide alone which may be attributed to Vit.D
supplementation has a potent stimulatory effect on bone formation through it was
effected on the activity of the hormones regulating Ca and P in blood concentrations
and parathyroid hormones to their normal concentrations.
Vit.D has a stimulatory effect on bone formation and mineralization thus vit.D
plays an vital role in the progress of the skeletal system. This result agrees with(31),
(32)who found that Vit.D is beneficial to body weight. This result is supported by
histological findings of parathyroid gland and kidneys after Vit.D supplmentation.
The enhancement of histological parathyroid gland which including chief cell is
responsible on secretion PTH as well as kidneys are responsible on excretion of Ca
and P by glomerular.
The Vit.D receptor and Vit.D metabolizing enzymes in male reproductive
system, particularly in the testis, suggests the happening of Vit.D synthesis and
regulation as well as function in the testis. The role of Vit D in the modulation of
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testis functions, including hormone production and spermatogenesis, has been
explored in animals. .
In fact, VitD was shown to be positively associated to sperm motility, and to
exert direct actions on spermatozoa, together with driven modulation of intracellular
Ca homeostasis and activation of molecular pathways involved in sperm motility, capacitation
and acrosome reaction. The present study confirmation on the role of Vit. D in male fertility,
by reporting trial studies in animals deal with the association connecting Vit. D and testis
role. This results are agreement with (33)who indicate by the estimation of both VitD and
the alternate marker 25-hydroxy-Vit.D3 circulating points, on semen quality and
fertility have been explored in vivo studies in animal models.
تقییم دور اضافة فیتامین د على بعض المعاییر الفسیولوجیة وفعالیة الغدة جار الدرقیة
ونوعیة السائل المنوی فی ذکور الأرانب المستحث فیھا نقص فیتامین د بواسطة
الفوروسیمید
ایات رحمن صبیح ، منى حمید السعید
فرع الفسلجة و الأدویة والکیمیاء الحیاتیة ,کلیة الطب البیطری,جامعة البصرة ,العراق.
الخلاصة
وی ائل المن صممت ھذه الدراسة لتقصی التأثیر العلاجی لفیتامین د على المعاییر الفسلجیة وصفات الس
ب ة الط وانی لکلی ت الحی ی البی ة ف ت الدراس د أجری امین د، وق ص فیت ا نق تحدث فیھ ب المس ور الأران ی ذک ف
ى وائیة إل ورة عش ارب بص ات التج مت حیوان ة ,قس ب المحلی ور الأران البیطری- جامعة البصرة على 30 من ذک
وم دة ٣٠ ی ل)لم یولوجی ( ٣م المحلول الفس ت ب البة عومل یطرة الس ة الس ی مجموع ى ھ امیع. الأول ة مج خمس
مس ة للش ر معرض ة غی والمجموعة الثانیة ھی السیطرة الموجبة وضعت الارانب فی اقفاص داخل غرفة مظلم
ص تحداث نق وم لاس دة ١٦ ی 0.5 ) لم mg/kgB.W.) furosemide ت ب ذلک عومل دودة وک ذیتھا مح وتغ
مس ة للش ر معرض ة غی ة مظلم ل غرف اص داخ ی اقف ب ف عت الاران ا وض ة ایض ة الثالث امین د ،و المجموع فیت
وم + ( 200000 دة ١٦ ی 0.5 ) لم mg/kgB.W.) furosemide ت ب ذلک عومل دودة وک ذیتھا مح وتغ
ر ة غی ة مظلم ل غرف اص داخ ی اقف ب ف کل ١٠ ایام لمدة شھر ،کذلک المجوعة الرابعة وضعت الاران (IU/2ml
وم + دة ١٦ ی 0.5 ) لم mg/kgB.W.) furosemide ت ب ذلک عومل دودة وک ذیتھا مح مس وتغ ة للش معرض
ة ل غرف اص داخ ی اقف ب ف عت الاران رة وض 100000 ) کل ١٠ ایام لمدة شھر بینما المجموعة الاخی IU/1m))
دة ١٦ 0.5 ) لم mg/kgB.W.) furosemide ت ب ذلک عومل مظلمة غیر معرضة للشمس وتغذیتھا محدودة وک
دل م ومع اس وزن الجس م قی ھ ت اء التجرب یوم + التعرض للشمس خمس ساعات فی الیوم لمدة شھر. بعد انتھ
اء تخراج الاعض ات لاس حیة بالحیوان م التض ک ت د ذل دم بع ات ال حب عین م س ات وت ع الحیوان الزیادة الوزنیة لجمی
م ی وزن الجس وی ف اض معن ک انخف ائج ان ھنال ت النت ات. بین ع الحیوان ن جمی وی م ائل المن المدروسة وجمع الس
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Testosterone ون امین د وھرم یوم وفیت تویات الکالس ی مس وی ف ومعدل الزیادة الوزنیة وکذلک انخفاض معن
ص ا نق تحث فیھ ب المس ا للأران ا وحرکتھ ة فعالیتھ ة وقل ف الحی دد النط ف وع وکذلک انخفاض العدد الکلی للنط
ائج ت النت مس بین عة الش ة لاش امین د والمعرض ة بفیت امیع المعالج یطرة والمج ة الس فیتامین د مقارنة مع مجموع
ذلک ایرود وک ون البراث فور وھرم ک والفس امض الیوری اتنین و ح ا والکری توى الیوری ی مس ة ف ادة معنوی ھناک زی
ب ذکور الاران ى ل ى والخص ة والکل ار الدرقی دة ج بة للغ جیة بالنس رات نس اک تغی وھن LH وFSH ونی ھرم
مس عة الش المستحث بھا فیتامین د. بینما اظھرت نتائج المجامیع المعاملة بفیتامین د و المجموعة المتعرضة لاش
ھناک تحسن واضح بکل المعاییر المذکورة سابقا وکذلک المقاطع النسجیة .
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