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ZANCO Journal of Pure and Applied Sciences
The official scientific journal of Salahaddin University-Erbil
ZJPAS (2016), 28 (5); 621-135
Histological Interfaces of Liver, Kidney and Cerebrum in Male Rats
exposed to Fluoxetine
Sarkawt H. Hamad1, Dlshad Hussein Hassan
1, Khder H. Rasul
2, Nazar Mohammed Shareef
Mahmood3
6- Biology Department, Faculty of Science, Soran University, Soran, Erbil, Kurdistan Region, Iraq.
2- Department of Biology, College of Science, Salahaddin University, Erbil, Kurdistan Region, Iraq.
3- Bjeel Preparatory school, Directorate of Education - Akre, General Directorate of Education – Duhok, Ministry of
Education- KRG-Iraq.
1. INTRODUCTION
Depression is a condition of unfavorable
mood and dislikes to do activities which lead to
abnormal behavior and aggressive feeling.
Depressed mood is a normal occurrence in
response to adversity in all individuals and it is
very common among persons who had a
trouble and disease (Johnstone et al., 2007).
The diagnoses such as Beck Depression
Inventory and Children’s Depression Inventory
have been used for assessment of depression
and severity of its symptoms (Biros et al.,
2008, Jang et al., 2016). In general, depressed
mood may not need a professional treatment,
but in most cases, the patients take treatments,
so that the production and using of
antidepressant such as selective serotonin
reuptake inhibitors are the most frequently
prescribed drugs to treat moody disease among
A R T I C L E I N F O
A B S T R A C T
Article History:
Received: 21/07/2016
Accepted: 61/08/2016
Published: 28/11/2016
Fluoxetine is an antidepressant of selective serotonin reuptake inhibitor
drug. It is used in treatment of depression, panic and anxiety. Also, it may
decrease the risk of suicide in those over the age of sixty-five years. The
current study was planned to investigate the histological effect of oral
administration of fluoxetine (10 mg/kg body weight/day) on liver, kidney and
cerebrum of male rats. Rats were divided randomly and equally into two
groups; control group (n=8) and fluoxetine-treated group (n=8). After one
month of administration; liver, kidney and cerebrum tissues would be taken to
make histological slides. Histological alterations were observed in liver such
as presence of few dead hepatocytes, congested blood vessels and
inflammatory cells. Kidney also showed histological changes; in the cortex
region include shrunken glomeruli tuft, hemorrhage and degenerated cells
while in the medullary region included thickened wall of tubules and some of
the epithelial tubule cells were degenerated. Alterations in the cerebrum were
the presence of dead neuronal cells in the second and third layers of grey
matter were due to fluoxetine. In conclusion, fluoxetine had a minute effect on
the histological structure of liver, kidney and cerebrum tissues.
Keywords:
Fluoxetine
Depression
Liver
Kidney
Cerebrum.
*Corresponding Author:
Sarkawt H. Hamad
Hamad, S. et al. /ZJPAS: 2016, 28(5): 126-135
127
them fluoxetine (Anderson, 2000, Inkielewicz-
Stepniak, 2011).
Fluoxetine is a selective serotonin reuptake
inhibitor and as a first line drug which is used
as a treatment of depression and many
neuropsychiatric disorders (Cipriani et al.,
2005, Wernicke, 2004). Serotonin in the
central nervous system act as neurotransmitter
included in variety of physiological and
behavioral functions and considered to have
an important role in the control of pain (Bardin,
2011).
Depression and drug using such as
fluoxetine gradually increases in all ages
especially among adults and teenagers in
Kurdistan region-Iraq. Fluoxetine is easily
available by the patients; it metabolizes in the
liver and excreted in the urine. This study was
planned to determine the histological effects of
fluoxetine on liver, kidney and cerebrum.
2. MATERIALS AND METHODS
2.1. Experimental Animals
Sixteen adult Wister male albino rats (8-10
weeks old) and weighing (200-270 gm) were
conducted in this study. They were housed in
plastic rat cages (56 x 39 x 19 cm) in groups
of eight rats per cage in a room with controlled
temperature of (22 ± 1 oC), 12 hours light and
12 hours dark by using an automated light-
switching device, in the animal house of
Biology department, Faculty of Science, Soran
University; under supervision and approval of
local scientific committee and animal care
rules. Rats were fed with standard laboratory
chow and allowed drinking water ad libitum.
2.2. Experimental design
Rats were divided randomly into two
groups; group 1: control group (n=8), group 2:
fluoxetine-exposed rats in which rats were
orally administrated with 10 mg/kg body
weight/day (n=8).
2.3. Anesthesia, dissection and removal of
the organs
At the end of the experiment, all rats were
anesthetized with intraperitoneal injection of a
mixture of Ketamine Hydrochloride (80 mg /
Kg) and Xylazine (12 mg / Kg), then liver,
kidneys and cerebrum were removed, cut into
smaller pieces (approximately 0.5cm in
thickness) in petri dish which contained a
fixative (Bouin’s fluid) and then transferred
into Bouin’s fluid for fixation.
2.4. Histological preparation (Paraffin
method)
Fixed tissues (liver, kidney and cerebrum)
were removed from Bouin’s fluid and
dehydrated by a serial concentration of ethanol
in ascending manner, infiltrated with paraffin
wax after the clearing. Paraffin wax also used
for making tissue blocks. The tissue blocks
were cut into four to six micrometer thick
paraffin sections by using rotary microtome
(Bright, MIC) and stained with hematoxylin
and eosin (H&E) (Bancroft et al., 1977).
Finally, light microscope (digital binocular
compound microscope 40x-2000x, built-in
3MP USB camera) was used to examine and
photo taken.
3. RESULTS AND DISCUSSION
Daily oral administration of fluoxetine for
one month caused several histological
alterations in liver, kidney and cerebrum of
rats.
3.1. Effect of fluoxetine on liver
Histological sections of liver control rats
showed normal hepatic architecture in which
Hamad, S. et al. /ZJPAS: 2016, 28(5): 126-135
128
many healthy hepatocytes were polyhedral in
shape appeared, exhibited centrally round or
ovoid shape nucleus and well- known plasma
membrane. Blood sinusoids appeared normally
and located between hepatocytes. Central vein
were normal (Fig. 3.1 and Fig. 3.2). The liver
sections of fluoxetine treated rats showed that
most of hepatocytes would be appeared as
normal and there is no obvious architecture
change except that a few of the hepatocytes
degenerated, a little inflammatory infiltration
of leukocytes and congested blood vessel
would be observed (Fig. 3.3 and Fig. 3.4).
Histopathological examination of the liver
revealed hepatic injury after fluoxetine
treatment (Yilmaz et al., 2016). Furthermore,
hepatocellular hydropic vacuolar degeneration,
portal area and lobular inflammation were
observed as a result of rat exposure to
fluoxetine which agree with our results (Özden
et al., 2005).
3.2 Effect of fluoxetine on kidney
The paraffin sections of the cortical region
showed well-designated glomeruli and kidney
tubules both proximal convoluted tubules and
distal convoluted tubules in the kidney of
control group rats (Fig. 3.5). The medullary
region of the control kidney rats also showed a
normal appearance of tubules as well as their
epithelial cells (Fig. 3.6). In contrast, fluoxetine
caused alterations in kidney rats of treated
group, the cortex region showed shrinking of
glomeruli, hemorrhage and dead cells (Fig. 3.7)
while in the medullary region, thickened the
wall of kidney tubules and degeneration of
some kidney tubule epithelial cells were
observed (Fig. 3.8). At doses of 3 and 10
mg/kg, the significant reductions in renal nerve
activity were observed especially 15 minutes
after the intravenous injection of fluoxetine
(Tiradentes et al., 2014). The hyponatremia
arose during fluoxetine antidepressant therapy
(ten Holt et al., 1996). Also, the sodium ions
level decreased, Potassium unchanged and
antidiuretic hormone remained unchanged,
whereas the AQP2 protein abundance and
water absorption in the inner medullary
collecting duct were increased (Moyses et al.,
2008).
3.3. Effect of fluoxetine on cerebrum
As shown in figure 3.9, the histological
slides showed that the cerebrum of control rats
have normal appearance of their structure such
as layers of grey matter and normal neuronal
cells, while dead pyramidal cells in second and
third layers of grey matter were observed in
the sections through the cerebrum of
fluoxetine-exposed rats (Fig. 8.10).It has been
suggested that fluoxetine cause increasing the
concentration of serotonin in synaptic cleft and
vasodilation of small cerebral arteries such as
branches of the anterior cerebral arteries which
induced by calcium channel openers (Ungvari
et al., 2000). In addition, the
electrophysiological studies have further
demonstrated that fluoxetine inhibits different
types of calcium channels in the neurons (Deak
et al., 2000). Fluoxetine at 0.03 mM enhanced
nicotine- and choline-induced relaxations in
which nicotine induced norepinephrine release
from cerebral perivascular sympathetic nerves
but vasorelaxation was blocked by higher
concentration of fluoxetine (>0.3 mM) that is
mean, the high concentration of fluoxetine
cause decrease neurogenic vasodilation while
low concentration of fluoxetine cause increase
neurogenic vasodilation (Chen et al., 2012).
Furthermore, fluoxetine alters the levels and
composition of brain GABA(A) receptors and
reduces the responsiveness of GABA(A)-R to
GABA-mimetic drugs such as pentobarbital
(Matsumoto et al., 2007).
Hamad, S. et al. /ZJPAS: 2016, 28(5): 126-135
129
Figure 3.1: Photomicrograph from liver a control rat showing the normal central vein (CV), normal hepatocytes and blood
sinusoids. H&E. 100X.
Figure 3.2: A magnified photomicrograph from liver control rat section showing normal appearance of hepatocytes (black
arrow) which polyhedral in shape, round or ovoid shape nucleus and well-defined plasma membrane. Blood sinusoids (S)
and central vein (CV) are normal. H&E. 400X.
CV S
CV
Hamad, S. et al. /ZJPAS: 2016, 28(5): 126-135
130
Figure 3.3: Photomicrograph from liver treated rat with fluoxetine showing that the hepatocytes will appear as normal and
there is no architecture change. A little inflammatory infiltration of leukocytes (black arrow) and congested blood vessel
(white arrow) will present due to fluoxetine drug. H&E. 100X.
Figure 3.4: A magnified histological section of liver treated rat with fluoxetine showing congested blood vessels (white
arrow) and leukocyte infiltration inflammation (black arrow). Most of hepatocytes show normal architecture and a few of
them degenerated. H&E. 400X.
Hamad, S. et al. /ZJPAS: 2016, 28(5): 126-135
131
Figure 3.1: Section through the cortex of the kidney of control rat showing normal glomeruli (black arrow) and kidney
tubules (white arrow). H&E. 400X.
Figure 3.1: Medullary region in the kidney of control rat having a normal appearance of tubules (T) H&E. 400X.
T
T
Hamad, S. et al. /ZJPAS: 2016, 28(5): 126-135
132
Figure 3.7: Photomicrograph from kidney cortex of treated rat with fluoxetine showing shrunken tuft of glomeruli (black
arrow), hemorrhagic area (H) and degenerated cell area (D). H&E. 400X.
Figure 3.8: Section through the kidney of fluoxetine treated rat showing the kidney tubules in the medullary region which
they had thickened wall (black arrow) and some of their cells were degenerated (white arrow). H&E. 400X.
H
D
Hamad, S. et al. /ZJPAS: 2016, 28(5): 126-135
133
Figure 3.9: Sections through the cerebrum of control rats. A) Grey matter of cerebrum shows different layers with normal
structure. H&E. 100X. B) Normal appearance of grey matter in which normal pyramidal cells (black arrow) and glial cells
(white arrow) were observed. H&E. 400X. C) Oil immersion magnification power showing normal pyramidal cells (black
arrow) and glial cells (white arrow) in cerebral cortex layer. H&E. 1000X.
A
A
C
B
A
Hamad, S. et al. /ZJPAS: 2016, 28(5): 126-135
134
Figure 3.10: Photomicrographs from cerebrum treated rats with fluoxetine. A) Grey matter of cerebrum which shows
layers. H&E. 100X. B) Dead cells (black arrow) were observed due to fluoxetine in second and third layers of grey matter.
H&E. 400X. C) High power magnification shows clearly dead cells (black arrow). H&E. 1000X.
4. CONCLUSIONS
The present work was designed to
determine the histological effect of fluoxetine
on liver, kidney and cerebrum of male rats. The
light microscopic examination showed that
orally administration of fluoxetine for a period
of one month had a bantam histological effect
of liver, kidney and cerebrum of male rats.
A
A
B
C
Hamad, S. et al. /ZJPAS: 2016, 28(5): 126-135
135
Conflict of Interest
There is no conflict of interest.
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