Global Journal of Pharmacology 7 (2): 123-132, 2013

ISSN 1992-0075
IDOSI Publications, 2013
DOI: 10.5829/idosi.gjp.2013.7.2.72171

Hepato-Renal and Hematological Effects of Diclofenac Sodium in Rats

1

Zeynab Kh. El-Maddawy and 1,2Ibrahim M. El-Ashmawy

Department of Pharmacology, Faculty of Veterinary Medicine, Alexandria University, Egypt

Department of Veterinary Medicine, Qassim Uviversity, Buridah, Qassim, 51452, Saudi Arabia

Abstract: The present study was carried out to evaluate the adverse effects of a daily single injection of
diclofenac sodium at different doses on some hematological, biochemical parameters and oxidative stress.
Histological changes in liver and kidney induced by diclofenac sodium were also investigated in male albino rats.
Forty five rats were randomly divided into 3 equal groups, the first group was given saline intramuscullary and
kept as a control.The second group recieved diclofenac sodium at a dose of 6.75 mg / kg b.wt. i. m. once/day.
The third group was given diclofenac sodium at a dose of 13.5 mg / kg b.wt. i. m. once/day. Diclofenac sodium
and saline were administered for two consecutive weeks. The obtained results showed that, diclofenac sodium at
dose of 13.5 mg / kg b.wt. induced a significant decrease in Hb, PCV, RBCs and WBCs values. Moreover there
were a significant decrease in serum total protein and albumin levels and significant increase in
aminotranseferases, alkaline phosphatase, urea and creatinine levels. In addition there were a significant
reduction in reduced glutathione(GSH) levels and significant increase in malondialdehyde (MDA) content in
liver and kidney homogenates. Histopathological alterations were found in livers and kidneys. However,
diclofenac sodium at dose of 6.75 mg / kg b.wt. induced non signaficant changes in the previous parameters. It
could be concluded that administration of diclofenac sodium at high dose induced some adverse effects on
hematological, biochemical, oxidative parameters as well as histology of liver and kidney. That could be
attributed to oxidative stress induced by the drug. However, these effects were reversible.
Key words: Diclofenac Sodium

Liver Kidney Histopathology

1.5 to 2.0 hours after
degenerative
rheumatic
ingestion
in
fasting
diseases,
as
well
as for
INTRODUCTION
subjects. Even though
painful conditions due to
inflammation
of
nonDiclofenac is a phenylacetic acid derivative that was diclofenac has a relatively
short
elimination
half-life
rheumatic
origin
and
acute
developed specifically as a non-steroidal antiattacks of gout [2,5]. It is
inflammatory (NSAID) drug. It competes with in plasma (1.5 hours), it
more effective in treating
arachidonic acid for binding to cyclo-oxygenase (COX), persists in synovial fluid.
Diclofenac
is
metabolised
ureteral colic than hyoscine
resulting in decreased formation of prostaglandins. This
in
the
liver
to
4butylbromide and has
decrease effect at least partly explains the mechanism of
hydroxydiclofenac
and
fewer side effects [6].
action of the drug [1,2]. Diclofenac has potency against
other
hydroxylated
forms,
Diclofenac has the capacity
COX-2 which is substantially greater than that of
after
glucoronidation
and
to treat uncomplicated
indomethacin, naproxen, or several other NSAIDs. The
sulphation
the
metabolites
urinary tract infections
selectivity of diclofenac for COX-2 resembles that of
caused by E. coli. [7]. It
celecoxib. In addition, it appears to reduce intra cellular are excreted in the urine
has also been shown to be
concentrations of free amino acid in leucocytes, perhaps (65%) and bile (35%) [1,2].
Diclofenac
has
analgesic,
effective
in
treating
by altering its release or uptake [3].Also diclofenac has
antinociceptive,
antipyretic
Salmonella
infections
in
anti-bacterial action shown by inhibition of DNA
and
anti-inflammatory
mice
[8]
and
is
under
synthesis [4]. Diclofenac is efficiently absorbed from the
investigation
for
the
gastrointestinal tract; peak plasma concentrations occur activities. It is used as
initial
therapy
for
treatment of
inflammatory
and
Corresponding Author: Zeynab

GSH

MDA

Khamis ELMaddawy,

Department of
Pharmacology,

Faculty of
Veterinary

Medicine
Alexandria
University,
Edfina-

123

RashidBehera,
P.O. Box:
22758,

Egypt.
Tel: +201007284
201, Fax:

+20452960450.

Global J. Pharmacol., 7 (2): 123-132,

2013

damage to
tuberculos outline the
is
[9].mechanisms
Diclofena of hepatoc
isrenal
associated toxicity of
the
drug.
with
Moreover,
severe
gastrointe we detected
whether the
stinal
toxicity adverse
effects
and
several induced by
adverse diclofenac
effects onsodium were
reversible or
lung,
hepatic not.
and renal
M
tissues
A
[10-13].
T
In
this
E
study the
R
effect of
I
diclofenac
A
L
sodium(in
S
jection) at
different
A
doses on
N
some
D
hematolo
gical,
M
E
biochemic
T
al
and
H
histopatho
O
logical
D
examinati
S
ons
of
liver andDrug:
kidney inDiclofenac
albino ratssodium
was
(Voltaren)
studied. ampoules
Also, we
each
one
investigat
contain 75
ed
the
mg
possible
diclofenac
linkages
sodium
is
between
produced by
drug
Novartis
induced
Pharma Co.
oxidative
Switzerland.
stress and
All
the
cell

diagnostic
kits assaying
hepatic and
renal
function
tests
were
obtained
from
Biodiagnostic
Company,
Egypt. Other
chemicals
used
obtained
from
ELGomhoria
Company,
Egypt.
Animals:
Forty
five
adult male
albino rats
(weighing
140 - 170 g
b. wt. and 18
weeks age)
were used.
The animals
were
obtained
from
a
closed
random bred
colony at the
Medical
Research
Institute of
Alexandria
University,
Egypt. The
rats
were
housed
in
plastic cages
(5
rats
/cage), with
free access
to
the
commercial
basal food
and water.
The standard
laboratory

diet wassodium and

purchased kept
as
from
control
Damanhu group.
r Feed Co.
(Behera, Group II:
Egypt). Rats were i.
All
m. injected
animals with
were
diclofenac
housed sodium at a
under thedose of 6.75
same
mg/kg b. wt.
environm
according to
ental
Paget
and
conditions
Barnes [14]
for
2
and
weeks
dissolved in
before
experime 2ml saline.
ntation for
Group III:
acclimatiz
Rats were i.
ation and
m. injected
to ensure
with
normal
diclofenac
growth
sodium at a
and
dose of 13.5
behavior.
mg/kg b. wt.
Experime in 2 ml
saline.
ntal
Diclofenac
Protocol: sodium and
After ansaline were
acclimatiz administered
two
ation, ratsfor
consecutive
were
weeks.
randomly
Five
divided
rats
from
into
3each treated
groups and control
(15 ratsgroup were
each):
killed after
2, 4 and 8
Group I:weeks from
Rats werebeginning of
i.
m.drug
and
injected vehicle
with
administrati
saline 2on.
ml/ kg b.
wt. as a
vehicle
for
diclofenac

Blood
Sampling:
At the end of
the
experiment,
rats
from
each group
were
anesthetized
with
light
ether
and
blood
samples
were drawn
from
their
retro-orbital
plexus
before killed
by
decapitation.
Two blood
samples
were
collected
from
each
animal. The
first sample
was
collected on
Disodium
salt
of
ethylene
diamine
tetra-acetic
acid (EDTA)
for
hematologic
al
studies.
The second
blood
sample was
collected
without
anticoagulan
t
for
obtaining
serum and
kept frozen
at
-20C
until
used
for
biochemical
analysis.
Blood
Analysis:

Hemoglo activities
were
bin
concentrat measured
ion wascolourimetri
determine cally
according to
d
according the method
to
thedescribed by
method Reitman and
described Frankel [17].
Alkaline
by
Benjamin phosphatase
[15]
by(ALP) level
was
using
Sahlis measured
hemocyto colourimetri
cally
meter,
Packed according to
the method
cell
volume described by
and
percent Kind
King
was
determine [18].Total
d
byprotein and
albumin
microhae
levels were
matocrite
measured by
technique
the
[16].
colourimetri
Erythrocy
c
method
tic
and
[19] , serum
total
globulin
leukocytic
level
was
counts
determined
were
by
performed
subtracting
by using
the albumin
Double
value from
improved
total protein
Neubauer value of the
hemocyto same sample
meter
as described
[16].
by
Coles
[20].Serum
Biochemi
urea activity
cal
was
Analysis:
measured by
Serum
the
alanine
enzymatic
aminotran
colourimetri
sferase
c method as
(ALT)
described by
and
Coulomb
aspartate
and Farreau
aminotran
[21]. Serum
sferase
creatinine
(AST)

activity was
measured by
the
colourimetri
c
kinetic
method as
described by
Husdan and
Rapoport
[22].
Antioxidant
Status and
Oxidative
Stress
Assays:
Specimens
from
liver
and kidney
of each rat
were
kept
frozen
at70C
for
assessment
of
malondialdh
yde (MDA)
and reduced
glutathione
(GSH)
contents.
After
ice
water
washing of
the tissues, a
part
from
each organ
separately
was weighed
and
homogenize
d
in
9
volumes
saline 0.9 %
in
Homogenize
r apparatus.
The
homogenate
was
collected
and kept at 70 o C. Lipid
peroxidation
as MDA was
measured
spectrophoto

metrically glutathione
after thelevel
was
reaction assayed
with
spectrophoto
thiobarbit metrically
uric acidaccording
according Sedlack and
to
theLindsay
method [24].
The
described method
is
by Placerbased on the
et al. [23].reductive
Reduced cleavage of
124

5,
5dithiobis- (2nitrobenzoic
acid)
(DTNB) by
sulfhydryl (SH) group to
yield
a
yellow
colour with
maximum
absorbance
at 412 nm.

Global J. Pharmacol., 7 (2): 123-132,

2013

different
Histopat changes of
hological melted
Examinat paraffin wax
at 60 C).
ion:
Following Paraffin
complete blocks were
by
necropsy cut
of
themicrotome
5
experime into
ntal malemicrons,
rats, smallthick
sections
fresh
specimens which were
from liverstained by
Haematoxyli
and
kidney n and Eosin
(H.E.)
were
collected according to
the method
and
rapidly described by
fixed inHarries [25]
10
%were
formalin examined.
solution
for
atStatistical
least 24Analysis:
hrs. afterResults were
that, thesestatistically
specimens analyzed by
one-way
were
processed analysis of
through variance(AN
OVA)follow
the
ed
by
convential
Duncans
paraffin
multiple
embeddin
range test.
g
Calculations
technique
were done
(dehydrati
using SAS
on
in
system [26].
ascending Data
are
grades ofpresented as
ethyl
means plus
alcohol, or minus the
clearing standard
in
error.
The
different minimum
changes level
of
of xylenesignificance
and
was set at p
embeddin 0.05.
g
in

RESULT
S
Hematologi
cal
Findings:
Administrati
on
of
diclofenac
sodium at a
dose of 6.75
mg/kg b.wt.
i.m. for 14
days to adult
male
rats
induced non
-significant
effect (P
0.05) on Hb,
PCV, RBCs
and WBCs
values
allover the
experimental
periods
compared to
control.
While there
was
a
significant
decrease(P
0.05) in their
values
in
rats treated
with
diclofenac
sodium
at
dose
13.5
mg/kg b.wt.
for 14 days
after 2nd and
4th weeks of
the
experiment.
At 8th weeks
from
drug
administrati
on all values
were
non
significant
as compared
with control
(Table 1).

Biochemi weeks from

cal
drug
Findings: administrati
The
on there was
results
nonshowed significant
that
effect
on
administr their values
ation ofas compared
rats withwith control
diclofenac (Table 2).
sodium at
Admini
dose 6.75stration of
mg/kg
diclofenac
b.wt. i.m.sodium
at
for
14dose of 6.75
days
mg/kg b.wt.
induced i.m. for 14
nondays to adult
significan male
rats
t effect oninduced nonserum
significant
ALT
,effect (P
AST
,0.05)
on
ALP, ureaserum total
and
protein
,
creatinine albumin and
levels
globulin
allover levels
the
allover the
experime experimental
ntal
periods
periods compared to
compared control.
to control.While there
While
was
a
there wassignificant
a
decrease (P
significan
0.05) in
t increase
serum total
(P 0.05)protein and
in
theiralbumin
values inlevels in rats
rats
treated with
treated diclofenac
with
sodium
at
diclofenac dose
13.5
sodium atmg/kg b.wt.
dose
offor 14 days
13.5mg/k at 2nd and 4th
g b.wt. forweeks of the
14 days atexpermint.
2nd and 4thAt 8thweeks
weeks offrom drug
the
administrati
experime on there was
nt. At 8th

nonsignificant
effect
on
their values
as compared
with control.
Also, there
was
non
significant
change
in
globulin
level in this
group (3) at
different
experimental
period
compared to
control
(Table 3).
Antioxidant
Status and
Oxidative
Stress
Assays:
The results
showed that
administrati
on of rats
with
diclofenac
sodium
at
dose
6.75
mg/kg b.wt.
i.m. for 14
days induced
nonsignificant
change on
liver
and
kidney LP
and
GSH
values
allover the
experimental
periods
compared to
control.
While, LP in
liver
and
kidney
tissues was
markedly
increased as
represented
by

consecu
Table 1: Effect tive
of i.m. days on

periods.
3

White blood cells count (10 / cmm)

injectio Hb g%,
n of

PCV

diclofe %,
nac

RBCs
sodium and
(6.75m WBCs
g and counts
13.5m of rats,
g/kg

at

b.wt.) differen
for 14 t

All values are

expressed as mean
Parameters
S.E. Number of rats in
Hemoglobin
each concentration
group is five. (g %)
Values with different
letters at the same raw
are significantly
Packed cell
volume (%)
different at P 0.05
(ANOVA with
Duncan's
multiple
Red blood
corpuscles
count (10
range test).

125

8Wk
2Wk

6.44 0.34 a
11.80 0.75 a

6.24

4Wk

11.100.40 a

11.3

8Wk

12.400.04a

11.9

11.2

Global J. Pharmacol., 7 (2): 123-132, 2013

Table 2: Effect of i.m. injection of diclofenac sodium (6.75mg and 13.5mg/kg b.wt.) for 14 consecutive days on serum ALT, AST, ALP, urea and creatinine
levels of rats, at different periods.

Groups
--------------------------------------------------------------------------------------------------------------------Parameters
Serum ALT activity (U/L)

Serum AST activity (U/L)

Duration

4Wk

44.402.43b

42.001.85b

8Wk

2Wk

2Wk
4Wk

42.001.13

127.406.83
130.007.50

128.406.88

85.58.24

84.84.24

90.226.33
30.14 0.27

28.80 0.29

8Wk

30.13 0.68

2Wk

0.87 0.01

0.87 0.01

2Wk
4Wk

4Wk

41.001.49
44.001.85

8Wk

Serum creatinine level (mg / dl)

39.80 1.72

8Wk

Serum urea level (mg / dl)

Diclofenac sodium (6.75mg/kg)

b

2Wk

4Wk
Serum ALP activity (U/L)

Control

Diclofenac sodium (13.5mg /kg)

83.202.83 a
74.43.11a
42.201.25a

130.008.13

168.405.48 a

129.005.26

160.003.42a

129.406.67

122.004.84a

95.52.24

195.29.24 a

80.63.24

168.18.24a

88.24.33

85.54.00 a

29.87 1.16

40.001.20 a

28.90 0.40

41.321.73 a

28.65 1.12

29.521.53a

0.93 0.02

1.280.01a

0.87 0.01

1.320.031a

0.90 0.01 a
0.90 0.01 a
0.920.02a
8Wk
All values are expressed as mean S.E. Number of rats in each group is five. Values with different letters at the same raw are significantly different at P
0.05 (ANOVA with Duncan's multiple range test).
Table 3: Effect of i.m. injection of diclofenac sodium (6.75mg and 13.5mg/kg b.wt.) for 14 consecutive days on serum total protein, albumin and globulin
levels, liver and kidney LP and GSH contents of rats, at different periods
Groups
---------------------------------------------------------------------------------------------------------------------Parameters
Serum total protein level (g /dl)

Duration

7.23 0.07

4Wk

7.170.03

7.340.02

3.06 0.03

3.04 0.01

3.12 0.01

4.17 0.04

4Wk

4.13 0.04

8Wk
2Wk

2Wk
4Wk
8Wk

Serum globulin level (g / dl)

Liver LP(nmol/MDA/g wet tissue)

Liver GSH (mol/g wet tissue)

7.430.08
7.260.06

6.960.04

Diclofenac sodium (2.6mg/kg)

6.360.03 b
6.200.06b
6.950.09a

3.00 0.01

2.130.02 b

3.040.02

2.140.01 b

2.950.03

2.990.01 a

4.43 0.07

4.230.07 a

4.21 0.07

4.060.05 a

4.22 0.01 a

4.01 0.06 a

3.950.02 a

5.06 0.11 b

5.020.03 b

8.150.06 a

4Wk

5.05 0.09 b

4.980.06 b

8. 500.04 a

8Wk

2Wk

2Wk
8Wk
2Wk
4Wk
8Wk

Kidney GSH (mol/g wet tissue)

4Wk
Kidney LP(nmol/MDA/g wet tissue)

Diclofenac sodium (1.3mg/kg)

a

2Wk
8Wk

Serum albumin level (g / dl)

Control

2Wk
4Wk

5.02 0.04

5.00 0.01

5.220.08 a

15.000.24

11.830.46 b

15.050.13

15.120.26

14.900.11

4.10 0.09

4.26 0.08

4.170.06

10.300.04

10.480.06

a
a

15.080.22

12.800.34 b

15.060.21

14.280.28 a

4.320.09

6.070.09 a

4.38 0.11

5.900.34 a

4.23 0.12

4.170.28 a

9.940.09

8.040.10 b

9.660.11

8.210.12 b

10.260.07
9.610.09
10.000.21 a
8Wk
All values are expressed as mean S.E. Number of rats in each group is five. Values with different letters at the same raw are significantly different at P
0.05 (ANOVA with Duncan's multiple range test).

126

Global J. Pharmacol., 7 (2): 123-132, 2013

Fig. 1: Photomicrograph of rat liver and kidney stained with HE: (A) Normal histologic architecture of liver of control rats
(Bar=500 microns). (B) Liver of rats treated with diclofenac sodium at dose of 6.75mg/kg b.wt. killed at 2 nd weeks
of the experiment: Few hepatocytes have mild hepatocellular hydropic degeneration (non-fatty type) (arrows)
(Bar=200 microns).(C)Liver of rats treated with diclofenac sodium at dose of 13.5mg/kg b.wt. killed at 2 nd weeks
of the experiment: Moderate diffuse hydropic degeneration and vacuolation (non-fatty type) ( Bar=200 microns).
(D) Liver of rats treated with diclofenac sodium at dose of 13.5mg/kg b.wt. killed at 4 th weeks of the experiment:
Periacinar necrosis (arrow heads) with mild to moderate infiltration of portal areas with mononuclear cells (arrow)
(Bar=500 microns). (E) Normal histologic architecture with normal proximal convoluted tubules lined by cuboidal
epithelium (arrows) of kidney control of rats (Bar=200 microns). F: kidney of rats treated with diclofenac sodium
at dose of 13.5mg/kg b.wt. and killed at 4 th weeks of the experiment: Several proximal convoluted tubules have an
attenuated and necrotic epithelium (arrows) (Bar=200 microns).
significantly decreased (P
on LP and GSH values as
compared with control
a significant increase (P 0.05) in MDA level in rats
0.05) at the same
(Table 3).
treated with diclofenac sodium at dose of 13.5mg/kg periods. At 8thweeks from
b.wt. for 14 days at 2nd and4th weeks of the experiment. drug administration there
GSH content in both tissues in that group were
was non-significant effect

127

Global J. Pharmacol., 7 (2): 123-132,

2013

week of the
Histopat experiment
hological the hepatic
Findings: tissues
appeared
Light
microscop nearly
normal
e
evaluation structure.
of
of liversLivers
in
therats treated
control with
diclofenac
rats
at
showed sodium
of
normal dose
morpholo 13.5mg/kg
for
gy of theb.wt.
hepatic successive
parenchy 14 days and
ma (Fig.killed at 2nd
week of the
1A).
Liver ofexperiment
showed
rats
treated moderate
diffuse
with
diclofenac hydropic
sodium atdegeneration
dose
ofand
6.75mg/k vacuolation
g b.wt. for(non-fatty
successiv type) (Fig.
e 14 days1C). At 4th
and killedweek, livers
rats
at 2nd or 4of
th
weeks oftreated with
that
dose
the
experime showed
periacinar
nt,
showed necrosis
with mild to
few
hepatocyt moderate
es haveinfiltration
of
portal
mild
hepatocell areas with
mononuclear
ular
hydropic cells(Fig.
degenerati 1D). At 8th
on(non- week of the
fatty type)experiment
the
same
(Fig.
1B),while lesions were
, at 8thpresent but

mild
in
nature.
Kidney of
rats
of
control
group
showed
normal
histologic
architecture
with normal
proximal
convoluted
tubules lined
by cuboidal
epithelium
(Fig.
1E).
However,
kidney
of
rats treated
with
diclofenac
sodium
at
dose
of
6.75mg/kg
b.wt.
and
killed at 2nd
week
of
experiment
showed mild
interstitial
inflammatio
n, while at
other periods
of
experiment
kidney
displayed
nearly
normal
structure.
Regarding to
kidney
of
rats treated
with
diclofenac
sodium
at
dose
of
13.5mg/kg
b.wt.
and
killed at 2nd

or
4ththerapeutic
weeks ofagents, used
for
the
the
experime treatment of
nt thererheumatic
were milddiseases,
interstitial because they
inflammat have
ion andanalgesic,
glomerulo antipyretic
anti-nephritis and
inflammator
with
actions.
several y
is
proximal There
considerable
convolute
in
d tubulesinterest
have anthe toxicity
attenuated of diclofenac
because of
and
necrotic its clinical
epitheliu use.
The
m
(Fig.
thfindings
of
1F). At 8
week ofthe present
investigation
the
experime showed that
nt, mildadministrati
of
interstitial on
inflammat diclofenac
at
ion andsodium
necrosis dose of 13.5
with nomg/kg b.wt.
evidence for
of
successive2
glomerulo weeks
to
-nephritis male
rats
were
(group
3)
detected caused
a
in
significant
examined impairment
kidneys. in liver and
kidney
DISCUSSI
functions at
ON
2nd and 4th
weeks
of
Nonexperiment.
Steroidal
As, serum
antiALT, AST,
inflammat
ALP, urea
ory drugs
and
are
the
creatinine
most
frequently activities
prescribed were
significantly

elevated
while total
protein and
albumin

levels
urine (65%)
and
bile
were
significan (35%) [2].
Therefore, it
tly
decreased is toxic to
and
compared liver
to
theirrenal cells
correspon [28].
Diclofenac
ding
values insodium
control induce a rise
in
liver
group
(group 1)functions
and
theand has also
been
other
treated reported to
one (6.75cause
hepatitis and
mg/kg
hepatotoxicit
b.wt.)
[29,30].
(groups y
Our
results
2).
are
However,
there wascompatible
with those of
non
significan Basavraj et
t changeal. [31] who
reported that
in
globulin administrati
of
level inon
diclofenac
all
groups. sodium (9.5
Activ mg/kg b.wt.)
ities
oforally for 28
in
AST anddays
ALT areSwiss albino
mice
most
commonl induced
y
usedsignificant
biochemic increase in
of
al markerslevels
AST,
ALT,
of
liver
damage total
bilirubin,
[27].
and
Diclofena urea
creatinine
c sodium
and
is
extremely significant
metabolis decrease in
ed in thetotal protein
liver, theand albumin
metabolit values.
es
areTolman,
excreted Kappus and
in
theSergio and

antonio
[30,32,33]
reported that
diclofenac
sodium
induced
hepatotoxic
and
nephrotoxic
effects
in
both humans
and
experimental
animals.
Moreover,
Tomic et al.
[13] showed
that
rats
treated with
diclofenac
sodium in a
dose of 8
mg/kg/day
during
7
days per os
induced high
mortality in
the animals
which may
be
a
consequence
of
the
microscopic
changes in
the liver and
its lowered
function.
Also,
Hussain et
al.
[34]
noticed that
serum
creatinine
and
urea
levels were
elevated in
four avian
species
treated with
diclofenac
sodium.
Similar
results were
observed in

rats,
in
rats
calves andtreated with
Japanese diclofenac.
quails
Also, Syed
[35-37]. et al. [40]
Also,
support the
Hickey et evidence of
al.
[38]renal
observed function
that
impairment
diclofenac by
is
adiclofenac
powerful
sodium and
nephrotox
diclofenac
icant
at
potassium as
100, 200
the
drugs
and 300
were
mg/kg as
increased
it increase
urea
and
urea level
creatinine in
in mice.
rabbits
In
addition,
The
our
mechanisms
results areof diclofenac
agree withinduced
those ofhepatic
Eldin et idiosyncratic
al.
[39]adverse drug
who
reactions
reported remain
marked largely
increase unknown
in urinary[29,41,42].
uronic
There
was
acid
evidence that
excretion toxicity was
128

related to the
drug
metabolism
and
was
reduced by
the addition
of
cytochrome
p-450
inhibitors to
the
culture
medium
[41,43-45].
Three
metabolites
of diclofenac
sodium are
reported to be
responsible
for
diclofenac
sodium
toxicity in the
liver, namely
4 hydroxy 3
diclofenac,
5 hydroxy 4
diclofenac
and
5
hydroxy
6
diclofenac
[46-48]. Both
the formation
of a toxic
metabolite
and covalent

Global J. Pharmacol., 7 (2): 123-132,

2013

most
binding ofimportant
the drugcause of the
to hepaticadverse
of
proteins effects
have beendiclofenac
invoked on liver and
to explainkidney
tissues
in
its
toxicity sensitive
persons or
[49].
Mitochon animal
species and
drial
damage potentially
during long
and
use
NADPH term
deficiency [10,11].
are also,
The
thought toimpairment
be
in the liver
responsibl and kidney
e
forfunction
diclofenac testes will be
sodium taken as a
hepatotox markers of
icity
their
[50,51]. injuries. Our
How findings
ever, Bothshowed that
hepatocell administrati
on of rats
ular
cytotoxici with
ty fromdiclofenac
at
diclofenac sodium
or
itsdose of 13. 5
metabolit mg/kg b.wt.
es is onlyfor
observed successive 2
in
vitroweeks
with largeinduced
concentrat marked
ions
ofincrease in
diclofenac lipid
that areperoxidation
product,
not
achieved malondialde
in
vivohyde (MDA)
in
[51,52]. content
and
Inhibition liver
kidney
of
prostaglan tissues and a
dins is thesignificant

decrease in
GSH value
in
both
tissues at 2nd
and
4th
weeks of the
experiment
compared to
control.
However,
there
was
non
significant
change
in
their values
in
other
treated
group (group
2)
as
compared
with control.
The
status
of
lipid
peroxidation
as well as
the reduction
of
GSH
altered
levels
of
certain
endogenous
radical
scavengers
is taken as
direct
evidence for
oxidative
stress
affecting
functional as
well
as
structural
integrity of
cell
and
organelles
membrane
[53,54].
There were
little studies,
reported that
, the adverse

effects ofof Hickey et

diclofenac al. [38] who
sodium issuggested
contribute that
d
todiclofenac
oxidative induced
stress
nephrotoxici
[55].
ty
may
Duri involve
production
ng
diclofenac of reactive
sodium oxygen
metabolis species
to
m
theleading
number ofoxidative
and
reactive stress
oxygen massive
species genomic
can
beDNA
increased. fragmentatio
n and these
These
free
products two
induced radical
prooxidati mediated
events may
ve
damage inultimately
translate into
renal
apoptotic
tissue.
cell death of
The
increase kidney cells
in vivo and
in
a
superoxid reveal
DNA-active
e
for
dismutase role
levels anddiclofenac.
MDA
Our
activity inhistopatholo
renal
gicl findings
tissue
in livers and
may
kidneys
indicate tissues
go
peroxidati side by side
ve
with
the
damage obtained
and causebiochemical
cell
alterations.
damage inHistopatholo
kidney gic
tissue
examination
[56,57]. s
showed
Our
severity of
results
the lesions
supported
increased
by those
with

increased the
dose of drug,
moderate
diffuse
hydropic
degeneration
and
vacuolation
and
periacinar
necrosis
with mild to
moderate
infiltration
of
portal
areas with
mononuclear
cells in the

examined rats
and
hepatic rabbits
tissues ofrespectively.
rats
Also,
treated Hussain et
with
al.
and
diclofenac Triebskorn
sodium atet al. [34,59]
dose
ofobserved
13.
5some
mg/kg
histopatholo
nd
b.wt. 2 gical
and
4thalterations in
weeks ofliver
and
the
kidney
of
experime rainbow
nt.
Thetrout
renal
(Oncorhync
lestions inhus mykiss)
the sameand broiler
rats
birds
and
showed pigeon
several respectively
proximal treated with
convolute diclofenac
d tubulessodium.
have an
In the
attenuated present
and
study,
necrotic administrati
epitheliu on
of
m
withdiclofenac
mild
sodium
at
interstitial dose
13.5
inflammat mg/kg b.wt.
ion
i.m. for 14
glomerulo days to adult
-nephritis. male
rats
These
induced
findings significant
agree withreduction on
those
Hb,
PCV,
obtained RBCs and
by
WBCs
Iskandar values at 2nd
et al. EL-and,4th
Hamamm weeks of the
y et al.;experiment
Hickey et compared to
al.
andcontrol.
Aydin
While, there
[11,
was
non
35,38,58] significant
in mice,effect
in

their values
in
rats
treated with
diclofenac
sodium
at
dose of 6.75
mg/kg b.wt.
as compared
with control.
These
results are
agree with
those
of
Basavraj et
al. [31] who
reported that
diclofenac
sodium (9.5
mg/kg b.wt.)
orally for 28
days
in
Swiss albino
mice
induced
highly
significant
decrease in
Hb,
PCV
values.
Similar
results also,
were
observed by
Shridar and
Narayanan
[36].
Change
s
in
hematologic
al
picture
conclude to
anemia and
it may be
due to loss
of
blood
during
gastrointesti
nal bleeding
induced by
diclofenac
sodium [31].
The
obtained
results

showed hy (non fatty

that, theretype) at 2nd
was
and
4th
significan weeks with
t
dosevery
mild
dependent Interstitial
adverse inflammatio
effects ofn of renal
diclofenac tissue at 2nd
sodium inweek of the
rats.
experiment.
Diclofena
Despite,
c sodiumhematologic
at a doseal,
of
6.75biochemical
mg/kg
and
b.wt. didhistopatholo
not inducegical
any
alterations
significan induced by
t
high dose of
alterations diclofenac
in
allsodium
studied (13.5mg/kg
parameter b.wt.) at 2nd
s alloverand 4th week
the
of
experime experiment,
ntal
the changes
periods were
very
except
mild
and
mild
reversible at
cellular 8th
week.
swelling These results
and
agree with
vacuolar those
of
hepatopat Radford et
129

al. [60].
In
conclusion,
diclofenac
sodium
at
high
dose
causes
alterations in
hematologic
al,
biochemical
parameters
and
histopatholo
gical
changes in
liver
and
kidney
of
male
rats.
These
adverse
effects may
be
contributed
to oxidative
stress
induced by
the
drug.
However,
the
toxic
effects
of
diclofenac
sodium
could
be
acute
or
reversible.

Global J. Pharmacol., 7 (2): 123-132,

2013

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