Possible Prophylactic Effects of Vitamin E or Lycopene Treatment on Renal Toxicity Induced by CCL Administration in Albino Rats

World Journal of Zoology 2 (2): 19-28, 2007
ISSN 1817-3098
© IDOSI Publications, 2007
Possible Prophylactic Effects of Vitamin E or Lycopene Treatment on
Renal Toxicity Induced by CCL Administration in Albino Rats
4
Karima M. Moawad
Zoology Department, Girls College for Arts, Science and Education, Ain Shams University, Egypt
Abstract: The main purpose of this study was to evaluate whether vitamin E (40mg/kg) or lycopene (1 mg / kg)
treatments for 4 weeks are effective in preventing renal toxicity in male rats due to carbon tetrachloride
(CCL 1ml / kg, 3 times weekly) administration for 2 wee
4:
ks. CCL administration induced a significant decrease








4
in serum total protein and albumin, a significant increase (P<0.001) in serum urea and creatinine as well as in
sodium concentrations, accompanied with a significant decrease in circulating calcium level as compared to
the control group. CCL4- treatment induced a significant increase (P<0.001) in kidney total cholesterol,
triglycerides, total lipids, lipid peroxidation (MDA) and total protein levels as compared to control rats.
Moreover, intoxication with CCL caused a significant reduction in ki
4
dney glutathione (GSH) content associated
with the inhibition of gamma-glutamyl-transferase (GGT) and superoxide dismutase (SOD) activities. On the
other hand, vitamin E or lycopene administration for 4 weeks (2 weeks prior to and 2 weeks during CCL4
administration) caused improvement in serum and kidney parameters. The efficiency of vitamin E or lycopene
treatment to alleviate the serum concentrations of total protein, albumin, urea and creatinine as will as sodium
level was provoked. The reduction in serum calcium was modulated. Also, attenuation in the kidney cholesterol,
triglycerides, total lipids and MDA levels and total protein content was recorded when compared to CCL4
group values. Moreover, administration of vitamin E or lycopene prior to and concomitant with CCL injection
4
induced marked protection as detected by significant elevation in kidney antioxidants GSH, SOD and GGT
levels. The present results indicate that treatment of rats with vitamin E or lycopene might have produced
amelioration in kidney tissue through protection lipids indices and oxidative stress parameters against
CCL4-induced nephrotoxicity. Therefore, vitamin E or lycopene treatments may have therapeutic values in
preventing CCL4-induced nephrotoxicity.
Key words: Vitamin E % lycopene % rats % kidney % CCL4
INTRODUCTION
dismutase (SOD) scavenger systems [7]. It has been
suggested that certain vitamins and antioxidant
Carbon tetrachloride (CCl4) a clear, heavy and
compounds may act as preventive or protective factors
nonflammable liquid is widely used in the dry-cleaning
such as vitamin E [8] and carotenoids (Alpha- carotene,
industry and is a highly toxic chemical agent. Thus,CCl4
beta – carotene, lycopene and lutein) [9]. Lycopene is the
is most widely used for experimental induction of
pigment that gives tomatoes their red color and is one of
hepatic cirrhosis [1-3]. Exposure to CCl results in hepatic
4
four main carotenoids normally found in human blood
steatosis, centrilobular necrosis and ultimately, cirrhosis
and tissue. There is about 5 mg of lycopene per 100 gram
in the liver and acute tubular necrosis in the kidney [4].
of ripe tomato fruit [10]. Although not considered an
CCl4 induces oxidative stress in many settings and it also
essential nutrient, research has shown that lycopene
inhibits the activity of antioxidant enzymes in renal tissue
may have various benefits for human health. Recently, it
[5]. Lipids peroxidation is a major mechanism by which
was found that lycopene is an antioxidant known to
free radical can induce tissue injury [6]. Against these
decrease the risk of age-related chronic diseases, such
types of oxidative injuries, tissues have a variety of
as cancer [11] and cardiovascular disease in women [12].
defense mechanisms including the non-enzymatic
The pathogenesis of carbon tetrachloride (CCl4)-induced
glutathione (GSH) and the enzymatic superoxide
renal dysfunction is not completely known. Although
Corresponding Author: Dr. Karima M. Moawad, Zoology Department, Girls College for Arts, Science and Education, Ain
Shams University, Egypt
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World J. Zool., 2 (2): 19-28, 2007
different experimental animal models have been utilized
C
CCL4 plus Lycop group: rats received CCL 3 times






4
in order to explain such pathophysiological state, none
weekly for 2 weeks through i.p. plus oral dose of
of them have completely explained the mechanisms
lycopene (1 mg/kg) for 4 weeks (2 weeks before and
involved. It may be due to the functional state of the liver,
2 weeks during CCL administration).
4
or renal injury may develop independently of hepatic
events [13].
The doses of CCL , Vit E and Lycopene used i
4
n
Therefore, the aim of this study is to evaluate
this study were based on the efficacy of these drugs
renal dysfunction established as a consequence of
in rats [5, 8, 9].
acute liver damage after Ccl administration in rat
4
s
At the end of experimental period, rats were dissected
and to observe the changes in the antioxidative
under anaethesia then blood samples were collected from
defense enzymes. Also, to investigate the possible
heart, centrifuged and the sera were separated and stored
prophylactic effect of lycopene against oxidative damage
at -20°C.
of CCl in rats, compared wi
4
th the well-known antioxidant
Samples, approximately 0.5 g from kidney tissue,
vitamin E.
were taken and homogenized in ice-cold distilled
water, centrifuged at 3000 r.p.m. for 15 minutes and the
MATERIALS AND METHODS
supernatant was separated and kept frozen at -20°C until
assayed.
Sixty male albino rats (Rattus norvigicus) weighing
about 100-120 g each were used in this experiment. The
Biochemical measurements:
animals were housed in the usual metal cages at room
temperature and fed commercial laboratory show (solid)
C
Total protein content was measured by the Biruet
and had free access to water.
method [14], while albumin content was assessed
using the method of [15].
The drugs used:
C
Serum urea level was determined according to the
method of [16].
C
CCL :
4 A colorless non-flammable liquid, of
C
Serum creatinine was determined using kits
molecular weight 153.84 was obtained from El-Nasr
purchased from Stanbio laboratory Co. USA.
Pharmaceutical Chemical Co., A.R.E.
C
C
Calcium and sodium contents were determined using
Vitamin E, lycopene and other chemicals used for
reagent kits of Quimica Clinica Aplicada S.A (QCA),
estimation of SOD and GSH were obtained from
Spain, through Gamma Trade Co. Cairo.
Sigma Chemical Co., St-Lowis. Mo., USA.
C
Total cholesterol and triglycerides (TG) were
estimated by using enzymatic colorimetric kits
The experimental procedures: The animals were
purchased from Stanbio laboratory Co. (USA),
randomly allocated to six groups each of ten rats:
through Gamma Trade Co. Cairo.
C
C
Total lipid was detected according to the method of
Control group: rats received orally saline solution
(0.2 ml).
White et al. [17].
C
C
Vitamin.E (Vit. E) group: rats received oral dose
Determination of malondialdehyde (MDA) a
(40 mg/kg) dissolved in saline solution twice a week
marker of lipid peroxidation was performed using
for 4 weeks.
thiobarbituric acid reacting substance (TBARS)
C
Lycopene (Lycop.) group: rats received oral dose
according to the method of Ohkawa et al. [18].
(1 mg/kg) dissolved in saline solution twice a week
C
Gamma-glutamyl trasferase (GGT) was determined
for 4 weeks.
using kit purchased from Stanbio Chemicals (USA)
C
CCL
through Gamma Trade Company, Cairo.
4 group: rats received CCL



4 (1 ml/kg) 3 times
weekly for 2 weeks through intraperitoneal injection
C
Determination of glutathione reduced form
(i.p.).
(GSH) was performed according to the method of
C
CCL
Beutler et al. [19].
4 plus Vit.E group: rats received CCL 3 times






4
weekly for 2 weeks through i.p. plus oral dose of
C
Superoxide dismutase (SOD) activity was
Vit. E (40 mg/kg) for 4 weeks (2 weeks prior to and
determined by the method of Minami and Yoshikawa
2 weeks during CCL administration).
[20].
4
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World J. Zool., 2 (2): 19-28, 2007
Statistical analysis: Data were performed using
In comparison to the control values, CCL4 - treated
Students "t" test according to the method of Hin and
rats showed a significant increase (P<0.001) in
Wetherill [21]. Analysis of variance (ANOVA) and
kidney total cholesterol (6.07±0.27 mg/g), triglycerides
Fisher's significant difference test were also used [22].
(17.69±0.62), total lipids (37.04±0.77) and MDA
P values of 0.05 or less were considered as statistically
(500.67±23.92) levels when compared to the control
significant.
values. Also, kidney total protein level increased (97.8%)
significantly in rats treated with CCL (Tablea 2 & 3).
4
RESULTS
The intoxication of rats with CCL caused
4
a
significant reduction (P<0.001) in kidney levels of GSH
The effects of vitamin E or lycopene on serum
(-48.11%), GGT (-28.42%) and SOD (-44.61%) as compared
parameters of male rats treated with CCL4 are shown
to control group (Table 3).
in Table (1). The present data indicate that CCL4
On the other hand, treatment of CCL4 intoxicated
administration for 2 weeks induced a significant
rats with vitamin E or lycopene for 4 weeks
reduction (P<0.001) in serum total protein (-34.88%)
caused improvement in serum and kidney parameters.
and albumin (-28.47%) and a significant increase (P<0.001)
The reduction in serum total protein was abolished
in urea and creatinine levels as compared to the
and recorded a significant elevation (P<0.001) after
corresponding control values. It induced also a significant
treatment with vit.E and lycopene as compared to CCL4
decrease (P<0.001) in circulating calcium level. These
group. Elevation in serum albumin level was also
changes were accompanied with a significant increase
recorded in those animals, compared to the same group.
(P<0.001) in serum sodium (27.83%) concentration
Furthermore, the treatment with tested drugs caused
compared to control group.
detectable decrease in serum urea and creatinine levels
Table 1: Changes in some serum parameters of rats treated with CCL4( 1 ml/kg) or /and vitamin E ( 40 mg/kg) and lycopene (1 mg/kg)
Parameters
--------------------------------------------------------------------------------------------------------------------------------------------------------------------------
T.Protein
Albumin
Urea
Creatinine
Calcium
Sodium
Groups
g/dl
g/dl
mg/dl
mg%
mg/dl
mEq/l
Co.
6.80±0.28
3.69±0.19
22.85±1.70
0.91±0.05
7.19±0.16
38.18±1.53
6.86±0.59
3.49±0.14
23.41±0.53
0.96±0.07
7.07±0.14
37.99±1.70
Vit.E
0.91%
-5.60%
2.45%
5.30%
-1.69%
-0.50%
N.S
N.S
N.S
N.S
N.S
N.S
6.97±0.30
3.39±0.16
23.06±0.88
1.07±0.06
6.85±0.26
39.33±1.76
Lycop.
2.58%
-8.12%
0.88%
17.73%
-4.78%
3.00%
N.S
N.S
N.S
N.S
N.S
N.S
4.43±0.318
2.64±0.18
36.03±0.72
2.07±0.07
3.51±0.16
48.81±1.71
CCL4
-34.88%
-28.47%
57.64%
126.69%
-51.12%
27.83%
a**
a**
a**
a**
a**
a**
6.70±0.31
3.23±0.11
22.84±1.54
1.08±0.14
5.15±0.27
39.51±0.83
CCL4+Vit.E
-1.38%
-12.50%
0.05%
28.89%
-28.40%
3.48%
b**
a*
b**
b*
a** b**
b**
6.78±0.56
3.09±0.08
21.99±0.81
1.26±0.21
4.72±0.17
40.66±0.81
CCL4+Lyco.
0.92%
-16.38%
-3.76%
38.03%
-34.29%
6.49%
b**
a*
b**
b*
a**b*
b**
ANOVA
P<0.001
P<0.001
P<0.001
P<0.001
P<0.001
P<0.001
F (5,30)
5.94
6.05
23.4
13.54
57.89
7.89
Data are given as: means± S.E for 6 rats/ group.
a : differs from control rats
b : differs from rats treated with CCL4
Significant at * p< 0.01 or **p<0.001
% of change from control.
N.S.: Non- significant
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World J. Zool., 2 (2): 19-28, 2007
Table 2: Changes in some kidney parameters of rats treated with CCL4
compared to CCL - treated
4
rats, but creatinine values were
(1 ml/kg) or /and vitamin E ( 40 mg/kg) and lycopene (1 mg/kg)
still higher than those of the corresponding control
Parameters
values (Table1).
-------------------------------------------------------------------------
Improvement of serum sodium was observed with
Cholesterol
T.G
T.lipids
MDA
vit E and lycopene treatments recording (3.48 % and
Groups
mg/g
mg/g
mg/g
nmol/g
Co.
1.57±0.10
11.81±0.42
23.37±1.51
257.83±13.70
6.49 %) respectively, compared to CCL group. Also,
4
a
Vit.E
1.64±0.09
11.14±0.52
24.20±0.74
246.83±47.48
marked improvement in the circulating level of calcium
4.03%
-5.69%
3.55%
-4.27%
was provoked in response to tested drugs compared to
N.S
N.S
N.S
N.S
those of CCL group (Table1).
4
1.55±0.13
10.60±0.69
24.06±1.13
263.50±12.58
In addition, treatments of vit. E and lycopene were
Lycop.
-1.38%
-10.19%
2.92%
2.20%
found to reduce the degree of kidney tissue damage in
N.S
N.S
N.S
N.S
biochemical findings (Tables 2 & 3). Also, these results
6.07±0.27
17.69±0.62
37.04±0.77
500.67±23.92
indicate that treatment with lycopene might have
CCL4
285.90%
49.7616.2%
58.47%
94.18%
produced amelioration in lipids indices and oxidative
a**
a**
a**
a**
2.86±0.29
10.85±0.30
23.64±1.53
330.83±15.19
stress parameters against CCL4-induced nephrotoxicity.
CCL4+Vit.E
81.97%
-8.13%
1.13%
28.31%
Vit E and lycopene therapies prevented significantly
a** b**
b**
b**
a** b**
lipids elevation, induced by CCL intoxication, results a
4
s
2.21±0.14
10.54±0.92
22.16±0.58
340.84±21.39
amelioration in kidney cholesterol (81.97%, 40.40%) and
CCL4+Lyco.
40.40%
-10.74%
-5.18%
32.19%
triglycerides (-8.13%, -10.74%) as well as total lipids
a** b**
a* b**
b**
a** b**
(1.13%, -5.18%), compared to CCL group (Table 2).
4
ANOVA
P<0.001
P<0.001
P<0.001
P<0.001
In comparison to CCL – intoxicated rats, Vit E
4
or
F (5,30)
88.48
34
25.27
14.15
lycopene treatment induced a significant decrease
Data are given as: means± S.E for 6 rats/ group.
(P<0.001) in both kidney MDA and total protein levels
a : differs from control rats
Significant at * p< 0.01 or
b : differs from rats treated with CCL4
**p<0.001
and a significant increase (P< 0.001) in kidney GSH
% of change from control.
N.S.: Non- significant
content and GGT and SOD activities compared to CCL4
group (Tables 2 & 3).
Table 3: Canges in some kidney parameters of rats treated with CCL4
(1 ml/kg) or / and vitamin E ( 40 mg/kg) and lycopene (1 mg/kg)
DISCUSSION
Parameters
T.protein
GGT
GSH
SOD
The health benefits of lycopene including the
Groups
mg/g
u/g
mg/g
u/g
antioxidative effects are still under evaluation. Vitamin E
Co.
105.02±6.09
22.85±1.70
171.56±18.73 69.48±1.99
96.82±2.70
23.41±0.53
174.68±8.03
70.50±2.16
is a well-known antioxidant. Thus the present study was
Vit.E
-7.81%
2.45%
1.82%
1.47%
carried out to investigate the protective effect of lycopene
N.S
N.S
N.S
N.S
against oxidative damage of CCl in rats, compared wit
4
h
98.79±1.73
23.03±0.88
162.90±6.72
75.73±0.97
the well-known antioxidant vitamin E.
Lycop.
-5.93%
0.88%
-5.05%
8.98%
The current data showed that administrations of
N.S
N.S
N.S
N.S
vitamin E or lycopene to normal rats for 4 weeks did not
207.72±11.91 16.36±0.932
89.02±1.39
38.49±2.17
exert significant changes in all the investigated parameters
CCL4
97.80%
-28.42%
-48.11%
-44.61%
in both serum and kidney tissue.
a**
a**
a**
a**
Versus to normal control rats, CCL in
4
jection induced
131.61±18.39 22.84±1.54
173.51±7.29
81.63±3.40
CCL4+Vit.E
25.33%
0.05%
1.13%
17.49%
significant decrease in both serum total protein and
b**
b**
b**
a** b**
albumin levels and a significant (P<0.001) increase in
138.66±9.97
21.99±0.81
168.16±7.43
77.77±2.15
serum urea and creatinine concentrations. These changes
CCL4+Lyco. 32.04%
-3.76%
-1.98%
11.93%
in urea and creatinine levels may indicate a reduction in
a* b**
b**
b**
a* b**
the glomerular filtration rate (GFR) as a result of CCL4
ANOVA
P<0.001
P<0.001
P<0.001
P<0.001
intoxication, since the serum concentration of these two
F (5,30)
16.88
5.51
11.73
47.81
parameters depends largely on the glomerular filtration
Data are given as: means± S.E for 6 rats/ group.
[23]. Confirming these results Ogeturk et al. [24] found
a : differs from control rats
Significant at * p< 0.01 or **p<0.001
that serum urea and creatinine and renal malondialdehyde
b : differs from rats treated with CCL4
N.S..: Non- significant
(MDA) levels were increased significantly in rats
% of change from control
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World J. Zool., 2 (2): 19-28, 2007
following CCl exposure and also caused prominen
4
t
free radical scavengers and could protect against
damage in the kidney tissue as compared to the control
lipid peroxidation [1], or may be due to the improvement
group. Also, Rincon et al. [13] and Huizar et al. [25]
in kidney function. Supporting this explanation
observed that tubular changes, glomerular
Karahan et al. [33] found that lycopene as a novel natural
hypercellularity and capillary obliteration were
antioxidant might have protective effects against
significantly higher in the CCl4-treated group than with
gentamicin-induced nephrotoxicity and oxidative stress
control rats followed by a decreased in the glomerular
in rats. Also, Dillioglugil et al. [34] suggested that
filtration rate.
kidney function appears to be improved by vitamin E
The pathogenesis of renal function alteration
supplementation due to its antioxidant and antithrombus
associated with CCl4-induced liver disease remains to be
action. Moreover, the present study revealed a decrease
elucidated. Rincon et al. [13] suggested that the effect of
in circulating calcium level accompanied with highly
CCl
4 on kidney structure and function depended on the
significant (P<0.001) increase in serum sodium
functional state of the liver. In a previous study, Abd El
concentration. The decrease in serum calcium post CCL4
Dayem and Moawad [1] observed liver dysfunction and
injection was also reported by Nakano et al. [35] who
tissue damage of CCL intoxicated rats. CCl
4
induced
found that in rats with both chronic non-cirrhotic liver



4
oxidative stress in many settings [26, 27]. It is metabolized
injury and CCl4-induced cirrhosis, tibial bone volume was
through the mitochondrial monooxygenase system
significantly lower than in controls and the osteoid
(P450 2E1). During metabolism, an unstable free radical,
volume decreased while the urinary calcium /creatinine
trichloromethyl, exits and is rapidly converted to
ratio increased. Moreover, albumin is considered as the
trichloromethyl peroxide [28, 29]. Consequently, the cell
main Ca+ carrier through blood [36], so the decrease in
membrane structure and membranes of intracellular
albumin level is postulated to be the main cause of serum
organelles are totally damaged. In addition, CCL induced
+
4
Ca reduction.
injury of the basement membrane [13] which, in turn, will
However, the increase in serum sodium may be a
increase its permeability and thus increases protein loss
result of inhibited kidney excretion by CCL4 intoxication.
in the urine. In this line, Ozturk et al. [30] suggested that
This is supported by the findings of Yu et al. [37] who
the alterations in kidney functions and structure
reported that liver cirrhosis is associated with enhanced
observed in CCl treated rats, probably occurred as
4
a
renal tubular sodium retention and they hypothesized
result of the lipid peroxidation and breakdown of the
that the mechanism of which is associated with increased
membrane structure. Therefore, the present findings are
expression of renal epithelial sodium transporters.
in accordance with Lin et al. [3] and Ogeturk et al. [24]
Also, Huizar et al. [25] studied bile duct ligation (BDL)
who found that CCL4 intoxication significantly decrease
of variable etiology, either surgical or induced by CCl4 in
serum total protein and albumin levels in rats.
rats and found that urinary volume and urinary sodium
On the other hand, vitamin E or lycopene
concentration were significantly decreased, while plasma
administration for 4 weeks (2 weeks prior to and 2 weeks
renin activity and concentration, serum creatinine and
during CCL administration) caused improvement in seru
4
m
BUN values increased after BDL in cirrhotic rats. Also,
and kidney parameters. The efficiency of vitamin E or
glomerular filtration rate was substantially decreased.
lycopene treatment to improve the serum concentrations
On the other hand, the present results indicated that
of total protein, albumin, urea and creatinine was
the treatment of CCL4 intoxicated rats with vitamin E or
provoked. It has been well established that vitamin E
lycopene caused improvement in serum calcium and
attenuates the hepatotoxicity induced by carbon
sodium levels as well as kidney total protein content,
tetrachloride [1, 31]. The esters of vitamin E and synthetic
compared to those of CCL4- treated group. These changes
vitamin E-like antioxidant have been also found to reduce
may be due to the protection against peroxidative injury
carbon tetrachloride-induced liver injury [8, 32]. Also, Kim
induced by reactive oxygen species (ROS), which is
[9] found that carotenoids (alpha-carotene, beta–carotene,
associated with the risk of osteoporosis and can be
lycopene and lutein) have protective effects on oxidant-
reduced by certain dietary antioxidants. Rao et al. [11]
induced injury of rat hepatocytes and suppression of lipid
suggest that the dietary antioxidant lycopene reduces
peroxidation. The increases observed in serum total
oxidative stress and the levels of bone turnover markers
protein and albumin levels may be due to the improvement
in postmenopausal women and may be beneficial in
in protein synthesis in the liver as a result of antioxidant
reducing the risk of osteoporosis. In this line, Choi and
effect of both vitamin E and lycopene, which act as a
Rhee [38] indicated that vitamin E supplementation in
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World J. Zool., 2 (2): 19-28, 2007
chronic cadmium-poisoned rats normalized renal
Another mechanism for increase kidney lipids
dysfunction, blood pressure regulation and the glomerular
suggest a novel renal uptake pathway for Liver-type
filtration rate (GFR).
fatty acid binding protein (L-FABP), a carrier of
It was observed in the present study that the
hydrophobic molecules, some of which may exert
administration of CCL induced a significant increas
4
e
nephrotoxic effects. These explanations agree with
(P<0.001) in kidney total cholesterol, triglycerides, total
Oyama et al. [45] who detected high levels of L-FABP in
lipids and lipid perioxidation (MDA) levels. These
the circulation and in the kidney of rats with CCl -induced
4
changes recorded displayed lower levels in kidney tissue
acute liver injury compared with those in the control rat
of rats treated with vitamin E or lycopene. The increments
by immunoblotting. These circulatory L-FABP was found
of kidney lipids recorded after CCL4 administration
to be filtered by glomeruli and internalized by proximal
are in agreement with the finding of Kamalakkannan
tubule cells -mediated endocytosis.
et al. [39] who found that the levels of lipids, cholesterol,
In the present study, a marked increase in MDA
triglycerides and free fatty acids were increased in
concentration (p<0.001) was observed in kidney of CCL4
plasma and tissues (liver, kidney, heart and brain) of rats
intoxicated rats, which reflect, increased oxidative stress
treated with CCL (3 ml kg 1
1
4
G wkG ). Also, they found
in kidney tissue. The increase in lipid peroxidation in
that phospholipid levels increased in plasma, heart and
kidney post CCL administration was also reported
4
by
brain but decreased in liver and kidney. The mechanism
Abraham et al. [26]; Ozturk et al. [30] and Yoshida et al.
of CCL4 action on kidney lipids is complex and has
[31]. Reactive oxygen species have been implicated in the
not been elucidated completely. The formation of lipid
pathogenesis of renal injury by direct cellular toxicity,
peroxides induced by carbon tetrachloride has been
partly through liberation of vasoconstrictor-bioactive
observed in many studies [5, 40], results in oxidative
lipids and inactivation of nitric oxide (NO) [30]. In
damage to the lipids and proteins of the kidney in rats
addition, oxidized LDL is injurious to renal tubular
[26]. Also, a decrease in the activity of enzymes
epithelial cells and may contribute to tubulo-interstitial
protecting from lipid peroxidation in the kidney was
disease [46].
provoked [41]. The abnormally high kidney cholesterol
Versus CCL treated rats, administration of vitami
4
n
level detected in the present experiment might be
E or lycopene revealed attenuation in the levels of kidney
explained by raised levels of serum LDL-fractions [42].
total protein, total lipids, cholesterol, triglycerides and
These fractions are cleared chiefly through specific
MDA levels.
binding to LDL-receptors demonstrated in the
There are several defense mechanisms that protect
glomerular tissue [43]. Therefore, the high lipids levels
living organisms against free radicals. Vit-E, a fat-soluble
detected in CCL4- treated rats may be associated with
vitamin, is believed to be the first line of defense against
the accumulation of filtered LDL- fractions in the
cell membrane damage due to peroxidation [38] and
glomeruli, leading to a reduction in the glomerular
oxidative modification of LDL [47]. In support of this
filtration surface area. On the other hand, the
notion, Reckelhoff et al. [48] and Kwag et al. [49] found
accumulation of triglycerides may be due to the
that kidney function appears to be improved by vitamin E
degradation of phospholipids as a result of the increase
supplementation due to its antithrombus action, which
in lipid peroxidation [39]. The blockade of hepatic
in turn controls the arachidonic acid cascade system
triglycerides secretion by CCL4 accounts for characteristic
and effective in the suppression of oxidative stress. Also,
fatty liver [1,44] which caused damage of kidney structure
Ardestani et al. [50] concluded that irradiated mice that
and function. Rincon et al. [13] suggest that the effect
received Vit-E were able to restore the changes of lipid
of Ccl4 on kidney structure and function depended on
peroxidation and lipid profile and tolerated biomembrane
the functional state of the liver.
damage provoked by 1.09 Gy for 3 days.
In this respect, recent studies have shown that
Bud et al. [51] reported that tomato lycopene, alone
cirrhotic rats with CCL intoxication exhibited a
4
significant
or in combination with other natural antioxidants, inhibits
decrease in mean arterial pressure followed by a
LDL oxidation, cholesterol synthesis and augments LDL
decreased glumerular filteration rate (GFR) [24, 25]. The
receptor activity in macrophages. These results were also
aforementioned reduction in GFR and renal dysfunction
confirmed by clinical trials [52]. In this line, Hosomi et al.
run in agreement with the increase in kidney total lipids,
[53] reported that HgCl -induced increase of lipi
2
d
cholesterol, triglycerides and total protein contents
peroxidation in kidney was prevented by lycopene. Also,
recorded in the present study.
Fuhrman et al. [54] suggested that lycopene may act as
24

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World J. Zool., 2 (2): 19-28, 2007
moderate hypocholesterolemic agents, secondary to their
one of the most potent singlet oxygen quenchers,
inhibitory effect on macrophage 3-hydroxy-3-methyl
which suggests that it may have comparatively stronger
glutaryl coenzyme A (HMGCoA) reductase, the rate
antioxidant properties than other major plasma
limiting enzyme in cholesterol synthesis, also to augment
carotenoids [63].
the activity of macrophage LDL receptors.
In this line, Bhuvaneswari et al. [64] suggest that
Moreover, The present study demonstrated that
lycopene-induced increase in the levels of GSH, the
CCL4 intoxication induces a highly significant reduction
enzymes of the glutathione redox cycle and phase II
in kidney GSH content associated with the inhibition
(glutathione S-transferase -GST) in the buccal pouch
of kidney GGT and SOD activities. These changes are in
mucosa treated with DMBA. Another possible
agreement with recent studies [7, 55, 56]. They observed
explanation for lycopene alleviation of the CCl -induce
4
d
that rats received CCL recorded decrease in the levels o
4
f
nephrotoxicity is its antioxidant effects. According,
GSH content and enzyme activities of SOD, glutathione
Hosomi et al. [53] reported that glutathione peroxidase
peroxidase (GSPx) and glutathione reductase (GR) in liver
and catalase activities were enhanced, while superoxide
and kidney in comparison with controls as well as a
dismutase activity was depressed in HgCl -treated rat
2
s
significant increase in MDA level in the kidney was
when compared to control and these effects were
recorded. The decrease in the activity of SOD in the
prevented by lycopene. Moreover, Atessahin et al. [65]
kidney of CCl -treated rats may be due to the increase
4
d
reported that lycopene administration produced
lipid peroxidation or inactivation of the enzyme by
amelioration in biochemical indices of nephrotoxicity in
cross-linking with MDA. This will cause an increased
both plasma and kidney tissues as MDA, GSH, GSH-Px
accumulation of superoxide radicals, which could further
and CAT activities when compared to Cisplatin, which
stimulate lipid peroxidation. This explanation runs in
induced nephrotoxicity and oxidative stress in rat.
agreement with the findings of Anandan et al. [57] who
Therefore, it is possible suggested that the positive
reported that depletion of GSH results in enhanced lipid
effect of lycopene on CCl -induced nephrotoxicity migh
4
t
peroxidation, which in turn causes increased GSH
be the result of its antioxidant effects.
consumption and with Naik and Panda [58] who found
that CCl4 significantly altered SOD, GSH-Px and catalase
CONCLUSIONS
(CAT) levels, indicating that oxygen radicals play an
important role in CCl -induced renal damage. The decline
4
The present study suggested that CCl causes rena
4
l
observed in kidney GGT enzyme in the present study post
damage in rats through enhanced lipid peroxidation
CCL4 administration is an indication of impaired GSH
and significantly altered GSH, SOD and GGT levels,
synthesis. GGT enzyme is the first enzyme in the
indicating that oxygen radicals play an important role in
degradation of GSH to its constituent amino acids, which
CCl -induced
4
renal damage. Vitamin E or lycopene
were introduced in the resynthesizing of GSH to replenish
treatment normalized these changes, but treatment with
the tissue stores [59]. At the molecular level CCl4 activates
lycopene lead to more protection in kidney tissue than
tumor necrosis factor (TNF)alpha and transforming
vitamin E. This protection was indicated by the highly
growth factors (TGF)-alpha and -beta in the cell,
increase of kidney GSH content as well as SOD and GGT
processes that appear to direct the cell primarily toward
activities.
self-destruction or fibrosis [29]. The increase in kidney
Therefore, vitamin E or lycopene treatments may
total protein observed in the present study confirm the
have therapeutic values in preventing CCL4-induced
latter suggestions.
nephrotoxicity.
Administration of vitamin E or lycopene prior to
and concomitant with CCL injection induced marke
4
d
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