Drug and Chemical Toxicology

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Protective effect of captopril against diazinon

induced nephrotoxicity and neurotoxicity via
inhibition of ROS-NO pathway

Milad Vahidirad, Milad Arab-Nozari, Hamidreza Mohammadi, Ehsan Zamani

& Fatemeh Shaki

To cite this article: Milad Vahidirad, Milad Arab-Nozari, Hamidreza Mohammadi, Ehsan Zamani
& Fatemeh Shaki (2017): Protective effect of captopril against diazinon induced nephrotoxicity
and neurotoxicity via inhibition of ROS-NO pathway, Drug and Chemical Toxicology, DOI:
10.1080/01480545.2017.1391830

To link to this article: http://dx.doi.org/10.1080/01480545.2017.1391830

Published online: 08 Nov 2017.

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 DRUG AND CHEMICAL TOXICOLOGY, 2017
https://doi.org/10.1080/01480545.2017.1391830

RESEARCH ARTICLE

Protective effect of captopril against diazinon induced nephrotoxicity and

neurotoxicity via inhibition of ROS-NO pathway
Milad Vahidirada, Milad Arab-Nozarib, Hamidreza Mohammadib, Ehsan Zamanib and Fatemeh Shakib
a
Department of Toxicology and Pharmacology, Faculty of Pharmacy, Pharmaceutical Sciences Branch, Islamic Azad University, Tehran, Iran;
b
Department of Toxicology and Pharmacology, Faculty of Pharmacy, Mazandaran University of Medical Sciences, Sari, Iran

ABSTRACT ARTICLE HISTORY

Diazinon (Dz) is a widely used insecticide. It can induce nephrotoxicity and neurotoxicity via oxidative Received 31 July 2017
stress. Captopril, an angiotensin-converting enzyme inhibitor, is known for its antioxidant properties. In Revised 25 September 2017
this study, we used captopril for ameliorating of Dz-induced kidney and brain toxicity in rats. Animals Accepted 8 October 2017
were divided into five groups as follows: negative control (olive oil), Dz (150 mg kg
1), captopril (60
Downloaded by [University of New England] at 10:55 08 November 2017

and 100 mg kg
1) and positive control (N-acetylcysteine 200 mg kg
1) were injected intraperitoneally KEYWORDS
30 min before Dz. After 24 h, animals were anesthetized and the brain and kidney tissues were sepa- Captopril; diazinon;
rated. Then oxidative stress factors were evaluated. Also, blood was collected for assessment of blood neurotoxicity; nephrotox-
urea nitrogen (BUN), creatinine (Cr) and nitric oxide (NO) levels. Dz significantly increased oxidative icity; oxidative stress
stress markers such as reactive oxygen species (ROS), lipid peroxidation, and protein carbonyl as well as
glutathione (GSH) oxidation in both tissues. Increased levels of the BUN, Cr and NO were observed after
Dz injection. Interestingly, captopril administration significantly decreased ROS production in both tis-
sues. Captopril significantly protected kidney and brain against lipid peroxidation and GSH oxidation.
Administration of captopril could markedly inhibit protein carbonyl production in kidney and brain after
Dz injection. Furthermore, captopril ameliorated the increased level of BUN, Cr and NO. These results
suggested that captopril can prevent Dz-induced oxidative stress, nephrotoxicity and neurotoxicity
because of its antioxidant activity.

Introduction
Diazinon (Dz) is an organophosphate (OP) insecticide that
widely used in agriculture and households products to con-
trol insects (Oruç and Usta 2007, Jafari et al. 2012).
Nowadays, pesticide poisoning is one of the most common
causes of poisoning in the world, as WHO reported about
3 million cases of pesticide poisoning per year which lead to
more than 250,000 deaths annually (Hamad et al. 2016).
Although OPs are primarily known as acetylcholinesterase
inhibitors, induction of oxidative stress by OPs has been
reported as one of the main mechanism of tissue toxicity


for OPs poisoning in both humans and animals (Colovi c et al.
2015).
Oxidative stress is the imbalance between the rate of
reactive oxygen species (ROS) production and the protective
effects of antioxidants (Ahangar et al. 2017).
Previous studies revealed the role of oxidative stress in the
pathogenesis of OPs-induced acute and chronic intoxication
in clinical and experimental studies (Giordano et al. 2007,
Lukaszewicz-Hussain 2010). Exposure to Dz could lead to ser-
ious histopathological lesions in various tissues such as kid-
ney, brain and liver (Tsitsimpikou et al. 2013, Boussabbeh
et al. 2016).
Nephrotoxicity of Dz via oxidative damage has been
well shown in previous studies (Yavuz et al. 2003,
Boroushaki et al. 2013). On the other hand, the brain is the
most vulnerable organ to xenobiotic-induced oxidative dam-
age because of high polyunsaturated fatty acids (PUFAs) con-
tent, low levels of antioxidants and also a high rate of
oxygen consumption (Pizzurro et al. 2014). Several studies
have pointed to the potential role of oxidative stress in OPs
neurotoxicity (Giordano et al. 2007, Lukaszewicz-Hussain
2010, Rashedinia et al. 2016).
Increased ROS production after Dz exposure can attack to
cellular macromolecules and leads to lipid peroxidation, pro-
tein oxidation and DNA damage (Rashedinia et al. 2016).
On the other hand, endogenous nitric oxide (NO) is formed
from L-arginine by nitric oxide synthetase (NOS) enzyme
(Tutanc et al. 2012). The excessive production of NO is linked
to the oxidative stress and tissue damage (Peresleni et al.
1996). The role of NO in Dz-induced toxicity showed by
numerous studies (Beydilli et al. 2015, Abdel-Daim 2016).
Failure of antioxidant defense system in attenuation of
oxidative damage leads to initiation of cell death signaling
and finally tissue toxicity (Rashedinia et al. 2016). Therefore,
compounds with radical scavenging or antioxidant properties
can be beneficial for ameliorating tissue toxicity of Dz
(Boroushaki et al. 2013, Salem et al. 2015).
Captopril is an angiotensin-converting enzyme inhibitor
that is frequently used to treatment of hypertension disease

CONTACT Fatemeh Shaki fshaki.tox@gmail.com Department of Toxicology and Pharmacology, Faculty of Pharmacy, Mazandaran University of Medical
Sciences, Sari, Iran
ß 2017 Informa UK Limited, trading as Taylor & Francis Group
 2 M. VAHIDIRAD ET AL.
(Hosseinimehr and Karami 2005). It was shown that com-
pounds with sulfhydryl group have antioxidant effects
(Banerjee et al. 2010, Kasperczyk et al. 2014, Ortiz et al. 2016).
On this basis, the radical scavenging effect of captopril is
probably linked to a terminal sulfhydryl group in its structure
(Hosseinimehr and Karami 2005).
Interestingly, several studies showed that captopril
increased enzymatic and non-enzymatic antioxidants levels
and also decreased lipid peroxidation, protein oxidation in
various animal models (Yavuz et al. 2003, Ghazi-Khansari et al.
2005, Kalia et al. 2007).
Thus, the aim of the current study was to test the effect of
captopril treatment against DZ-induced nephrotoxicity and
neurotoxicity, focusing on its inhibitory effects on oxidative
stress.
Downloaded by [University of New England] at 10:55 08 November 2017
Methods
Animal treatment
Male Wistar rats (200–250 g) were kept in an air-conditioned
room with a controlled temperature of 22 ± 2  C and main-
tained on a 12:12 h light cycle with free access to food and
water. All experimental procedures were conducted according
to the ethical standards and protocols approved by the
Committee of Animal Experimentation of Mazandaran
University of Medical Sciences, Sari, Iran. All efforts were
made to minimize the number of animals and their suffering.
Animals were randomly divided into five groups (n ¼ 30)
and the groups were as follows: negative control group
(received olive oil), DZ group (received 150 mg kg
1 Dz intra-
peritoneally), captopril groups (that received 60 and 100 mg
kg
1 captopril intraperitoneally) and positive control group that
received N-acetylcysteine (NAC) (200 mg kg
1). After 30 min, DZ
(150 mg kg
1) that previously diluted with olive oil, injected to
captopril and NAC groups intraperitoneally. After 24 h, rats
were anesthetized and their brain and kidney tissues were
removed. After homogenization of tissues, oxidative damage
factors such as protein carbonyl, malondialdehyde (MDA), gluta-
thione (GSH), and reactive oxygen spices (ROS) were evaluated.
Furthermore, blood samples were collected for blood urea
nitrogen (BUN) and creatinine (Cr) and NO levels assessment.
Measurement of protein concentration
Protein content was determined in brain and kidney tissues
with Bradford method. Bovine serum albumin was used as a
standard, homogenate samples mixed with Coomassie blue
and after 10 min, absorbance was determined at 595 nm by
spectrophotometer (Parving et al. 2006).
Quantification of ROS level
The ROS level measurement was performed using dichloro-
dihydro-fluorescein diacetate (DCFH-DA) as an indicator.
Briefly, DCFH-DA was added (final concentration, 10 lmol l
1)
to samples (1 mg protein ml
1) and then incubated for
10 min. The amount of ROS generation was determined
through a Shimadzu RF5000U fluorescence
spectrophotometer at 485 nm excitation and 520 nm emission
wavelength the results were expressed as fluorescent inten-
sity per 1 mg protein (Gao et al. 2009).
Measurement of GSH content
GSH content was determined using 5,50 -dithiobis-(2-nitroben-
zoic acid) (DTNB) as the indicator and the developed yellow
color was read at 412 nm on a spectrophotometer (UV-
1601 PC, Shimadzu, Japan). GSH content was expressed as n
mol l
1 (Sadegh and Schreck 2003).
Measurement of lipid peroxidation
The content of MDA was determined using dithiobarbituric
acid (TBA) as an indicator. Briefly, 0.25 ml sulfuric acid
(0.05 mol l
1) was added to 0.2 ml samples (1 mg protein
ml
1) afterwards, with the addition of 0.3 ml 0.2% TBA. All
the microtubes were placed in a boiling water bath for
30 min. At the end, the tubes were shifted to an ice-bath and
0.4 ml n-butanol was added to each tube. Then, they were
centrifuged at 3500  g for 10 min. The amount of MDA
formed in each of the samples was assessed by measuring
the absorbance of the supernatant at 532 nm with an
ELISA reader (Tecan, Rainbow Thermo, and Austria).
Tetrametoxypropane was used as standard and MDA content
was expressed as l mol l
1 (Zhang et al. 2008).
Measurement of protein carbonyl
Determination of protein carbonyl by spectrophotometric
method, briefly 200 ll of brain and kidney tissues are needed
to homogenate. Samples are extracted in 500 ll of 20% (w/v)
TCA. Then, samples placed at 4  C for 15 min. The precipitates
are treated with 500 ll of 0.2% 2,4-dinitrophenylhydrazine
(DNPH) and 500 ll of 2 mol l
1 HCL for the control group, and
samples are incubated at room temperature for 1 h with vortex-
ing at 5 min intervals. Then proteins are precipitated by adding
55 ll of 20% TCA. The microtubes are centrifuged and washed
three times with 1000 ll of the ethanol-ethyl acetate mixture.
And the microtubes are dissolved in 200 ll of 6 mol l
1 guan-
idine hydrochloride. The carbonyl content is determined by
reading the absorbance at 365 nm wavelength (Aebi 1984).
Measurement of nitric oxide
NO was evaluated using the commercial kits based on the
Griess reagent. In this method, sulfanilic acid is quantitatively
converted to a diazonium salt by reaction with nitrite in acid
solution. The diazonium salt is then coupled to N-(1-naphthyl)
ethylenediamine, forming an azo dye that can be spectro-
photometrically quantitated based on its absorbance at
548 nm (Shahani et al. 2016).
Statistical analysis
Results are presented as mean ± SD. All statistical analyses
were performed using the SPSS software, version 14
(Chicago, IL). Statistical significance was determined using the

one-way ANOVA test, followed by the post-hoc Tukey test.
Statistical significance was set at p < 0.05.
Results
Effects of captopril on ROS formation in rat brain and
kidney
ROS formation is an indicator of oxidative stress that signifi-
cantly (p < 0.001) were increased in rats that received DZ as
compared to negative control group in both kidney and brain
tissues. Data showed that ROS formation markedly was
decreased in groups that received both doses of captopril (60
and100 mg kg
1) as compared to the group that received DZ
in brain tissue. While in kidney tissue, administration of cap-
topril at the dose of 100 mg kg
1 inhibited Dz-induced ROS
formation as well as NAC group (p < 0.001) (Figure 1).
Downloaded by [University of New England] at 10:55 08 November 2017
Effects of captopril on lipid peroxidation in rat brain
and kidney
One of the end products of lipid peroxidation is MDA, and
elevation of MDA is known as an important marker for oxida-
tive stress. As shown in Figure 2, MDA level was increased
(p < 0.001) in rats that received DZ as compared to negative
control group in both kidney and brain tissues (Figure 2).
Data showed that MDA level was significantly decreased
in the group that received a high dose of captopril
(100 mg kg
1) compared to the group that received DZ in
both tissues as well as NAC group.
Effects of captopril on GSH concentration in rat brain
and kidney
Generally, imbalance between ROS and antioxidants levels
such as GSH caused oxidative stress-induced organ damage.
Figure 1. Effects of captopril on ROS formation in rat brain and kidney. ROS formation was determined in N.C (negative control), Dz (diazinon), Dz þ CP (captopril)
and Dz þ NAC (positive control) groups in the (A) brain and (B) kidney tissue, using DCFH-DA as described in ‘Methods’ section. Values represented as mean ± SD
(n ¼ 6). aaaSignificantly different compared to the NC group (p < 0.001), bbsignificantly different compared to Dz group (p < 0.01), bbbsignificantly different compared
to Dz group (p < 0.001), csignificantly different between two captopril groups (p < 0.05), ddsignificantly different compared to NAC group (p < 0.01), ns: non-
significant.
DRUG AND CHEMICAL TOXICOLOGY 3
The result showed that GSH levels significantly decreased in
rats that received DZ compared to negative control group in
both tissues. In addition, GSH concentration was increased
(p < 0.05) in the brain of rats that received 100 mg kg
1 of
captopril as compared to DZ group (Figure 3(A)). While GSH
levels elevation were not significant after administration of
both doses of captopril (60 and 100 mg kg
1) in kidney tissue
(Figure 3(B)).
Effects of captopril on protein carbonyl in rat brain and
kidney
Protein carbonyl was increased (p < 0.001) in the group that
received DZ as compared to negative control group in both
brain and kidney. As shown in Figure 4, the elevation of pro-
tein carbonyl content was significantly decreased in group
that received a high dose of captopril (100 mg kg
1) as com-
pared to group that received DZ in both brain and kidney tis-
sues (Figure 4).
Effects of captopril on nitric oxide levels in rat blood
serum
As shown in Table 1, NO generation significantly increased
after Dz injection. On the other hand, pretreatment with cap-
topril led to a reduction of NO production in serum (Table 1).
Effects of captopril on BUN and creatinine in rat blood
serum
According to Table 2, DZ injection was associated with signifi-
cant raise in serum levels of BUN and Cr, which can be indi-
cators of kidney damage. Administration of captopril at the
 4 M. VAHIDIRAD ET AL.
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Figure 2. Effects of captopril on lipid peroxidation in rat brain and kidney. Lipid peroxidation was determined in N.C (negative control), Dz (diazinon), Dz þ CP (cap-
topril) and Dz þ NAC (positive control) groups in the (A) brain and (B) kidney tissue, using TBA as described in ‘Methods’ section. Values represented as mean ± SD
(n ¼ 6). aaaSignificantly different compared to the NC group (p < 0.001), bsignificantly different compared to Dz group (p < 0.05), bbsignificantly different compared
to Dz group (p < 0.01), ns: non-significant.

Figure 3. Effects of captopril on GSH concentration in rat brain and kidney. GSH concentration was determined in N.C (negative control), Dz (diazinon), Dz þ CP
(captopril) and Dz þ NAC (positive control) groups in the (A) brain and (B) kidney tissue, using DTNB as described in ‘Methods’ section. Values represented as
mean ± SD (n ¼ 6). aaSignificantly different compared to the N.C group (p < 0.01), aaasignificantly different compared to the N.C group (p < 0.001), bsignificantly dif-
ferent compared to Dz group (p < 0.05), ns: non-significant.

dose of 100 mg kg
1 before Dz exposure could significantly
ameliorate these parameters as well as NAC group (Table 2).
Discussion
Pesticide poisoning is a concern for WHO that estimated 3
million poisoning cases per year. Dz is one of the most com-
monly used pesticide in the world (Hamad et al. 2016), that
notable cases of poisoning were reported worldwide. Dz poi-
soning leads to damage to several organs in the body,
including kidney, brain and liver (Tsitsimpikou et al. 2013,
Boussabbeh et al. 2016). Despite its major inhibitory effect on
acetylcholinesterase, it is observed that Dz exposure can lead
to organ toxicity via induction of oxidative stress (Yavuz et al.
2003, Lukaszewicz-Hussain 2010, Tsitsimpikou et al. 2013). So,
it is necessary to identify the effective and beneficial com-
pounds to the prevention of this unpleasant effect.
In our study, Dz administration led to increase of oxidative
stress markers such as ROS, lipid peroxidation, protein car-
bonyl and GSH oxidation in both kidney and brain tissues.
Induction of oxidative stress by Dz was shown by previous
studies that reported GSH oxidation (Ghazi-Khansari et al.
2005, Heo et al. 2008) and MDA elevation (Boroushaki et al.
2013, Abdel-Daim 2016) in the brain and kidney tissues
of rats after exposure to Dz, which confirmed the data of
our study.

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Figure 4. Effects captopril on protein carbonyl in rat brain and kidney. Protein carbonyl was determined in N.C (negative control), Dz (diazinon), Dz þ CP (captopril)
and Dz þ NAC (positive control) groups in the (A) brain and (B) kidney tissue, using DNPH as described in ‘Methods’ section. Values represented as mean ± SD
(n ¼ 6). aaaSignificantly different compared to the N.C group (p < 0.001), bsignificantly different compared to Dz group (p < 0.05), bbsignificantly different compared
to Dz group (p < 0.01), dSignificantly different compared to NAC group (p < 0.05), ns: non-significant.
Table 1. Effect of in vivo administration of captopril and
diazinon on serum nitric oxide (NO) concentration in rats.
Animal groups NO (mM)
N.C 5.20 ± 0.1
Dz 79.53 ± 11.91aaa
Dz þ CP (60 mg kg
1) 53.38 ± 7.1b
Dz þ CP (100 mg kg
1) 27.36 ± 7.8bbb,c
Dz þ NAC (200 mg kg
1) 33.43 ± 11.25bbb
N.C: negative control; Dz: Diazinon; Dz þ CP: captopril;
Dz þ NAC: positive control groups.
Values are expressed as mean ± SD for six rats in each
group.
aaa
Significantly different compared to the N.C group
(p < 0.001), bsignificantly different compared to Dz group
(p < 0.05), bbbsignificantly different compared to Dz group
(p < 0.001), csignificantly different compared to (Dz þ CP
60 mg kg
1) group (p < 0.05).
Because of high lipid content and also the low capacity of
antioxidants, brain is one of the most susceptible organs to
oxidative damage (Pizzurro et al. 2014). In addition, free radi-
cals can disrupt kidney function by the same way
(Tsitsimpikou et al. 2013).
So, compounds with antioxidant properties may be benefi-
cial to prevent oxidative damage of Dz. Previous studies
showed the protective role of antioxidant agents against Dz
toxicity. For example, aqueous extracts of Matricaria Recutita
L. and Asparagus officinalis could ameliorate Dz-induced tox-
icity through the MDA level reduction and elevation of GSH
content (Sulak et al. 2005). Also, pomegranate seed oil
reduced Dz-induced nephrotoxicity via inhibition of oxidative
stress in the kidney of rats (Boroushaki et al. 2013).
It is imagined that Dz toxicity might be due to binding
ability of Dz to sulfhydryl groups of GSH in the tissues (Kalia
et al. 2007). Several studies indicated that compounds with
sulfhydryl groups in their structures can bind to free radicals
and scavenge them and finally, reduce their destructive
effect. For example, Oksay et al. showed that NAC can
DRUG AND CHEMICAL TOXICOLOGY 5
Table 2. Effect of in vivo administration of captopril and diazinon on blood
urea nitrogen (BUN) and serum creatinine levels in rats.
Animal groups BUN (mg/dl) Creatinine (mg/dl)
N.C 23 ± 1.2 0.6 ± 0.08
Dz 68 ± 6a 1.23 ± 0.18a
Dz þ CP (60 mg kg
1) 46 ± 5 0.97 ± 0.1
Dz þ CP (100 mg kg
1) 33 ± 3b 0.65 ± 0.15b
Dz þ NAC (200 mg kg
1) 28 ± 4b 0.71 ± 0.13b
N.C: negative control; Dz: Diazinon; Dz þ CP: captopril; and Dz þ NAC: positive
control groups.
Values are expressed as mean ± SD for six rats in each group.
a
Significantly different compared to the N.C group (p < 0.05),
b
significantly different compared to Dz group (p < 0.05).
attenuate Dz-induced oxidative stress in the testis tissue of
rat by elevation in GSH level and reduction of MDA content
(Oksay et al. 2013). In another study, NAC could prevent GSH
depletion, lipid peroxidation and caused a significant increase
in endogenous antioxidant levels in liver oxidative damage
induced by malathion (Lasram et al. 2014).
Captopril is an angiotensin-converting enzyme inhibitor
that recently gained attention because of its antioxidant
activity (Ghazi-Khansari et al. 2005, Kalia et al. 2007). The pro-
tective effect of captopril in pathological conditions that oxi-
dative stress is involved were shown in previous studies
(Yavuz et al. 2003, Ghazi-Khansari et al. 2005, Kalia et al.
2007). For example, Ghazi et al. showed that captopril could
prevent liver oxidative damage induced by paraquat (Kalia
et al. 2007).
It is suggested that Dz toxicity is due to generation of free
radicals such as hydroxyl, superoxide and etc. (Ghazi-Khansari
et al. 2005). In this study, we showed that captopril could
prevent ROS production in both kidney and brain tissues.
Free radicals can attack to lipid membranes because of
the PUFAs available in the cell membranes, are very suscep-
tible to oxidation by ROS. Lipid peroxidation is a harmful out-
come of free radicals that can result in cell or organelle
 6 M. VAHIDIRAD ET AL.
membrane damage (Noctor et al. 2015). Our research indi-
cated that administration of captopril caused a significant
decrease in MDA level in both tissues.
Also, ROS can target other cellular macromolecules such
as proteins. This damage is generally associated with
increased level of carbonylated proteins that are good
markers for protein oxidation (Sharma et al. 2012).
Captopril could reduce the protein carbonyl levels in both
kidney and brain tissues. It means that probably captopril has
a potential effect of inhibition of protein oxidation induced
by Dz.
GSH is a tripeptide that has an undeniable role in the anti-
oxidant defense system by direct interaction with ROS. GSH
depletion may occur in various oxidative situations (Lushchak
2012).
In this study, Dz-induced GSH depletion was improved in
both tissues after administration of captopril.
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Role of NO in oxidative damage was shown in previous
studies (Naseh and Vatanparast 2014, Beydilli et al. 2015).
Furthermore, several studies showed that elevation of NO
level occurred after Dz exposure (Bedilli et al. 2015, Abdel-
Daim 2016). In our study, this elevation was seen, too. Some
antioxidant compounds can prevent NO generation and
inhibit its disruptive effects. For example, curcumin as an
herbal antioxidant could reduce NOS activity and NO produc-
tion (Bengmark 2006). Also, the previous study showed that
captopril could reduce the expression of NOS which led to
decrease in NO concentration. For example, Uhlenious et al.
demonstrated the protective role of captopril against NO-
induced renal dysfunction (Uhlenius et al. 1999). In this study,
captopril decreased the level of serum NO that probably is
due to its inhibitory effect on NOS.
Free radicals can cause leakage of the cell membranes by
attacking to the PUFA present in membrane structures and
lead to necrosis (Standman and Levine 2000). As shown in
the result, the levels of BUN and Cr increased significantly
after Dz administration.
Interestingly, after treatment with captopril, elevated levels
of BUN and Cr were decreased that confirmed the role of oxi-
dative damage in Dz-induced tissue toxicity.
As mentioned above, the antioxidant activity of captopril
is related to thiol groups in its structure that allows captopril
to prevent thiol group oxidation or its ability to direct bind-
ing to ROS and other free radicals. This effect finally could
result in amelioration of damage to cellular macromolecules
such as membrane lipids and proteins.
Taken together, captopril can inhibit cell membrane dam-
age and cell death which finally reduced tissue toxicity (Ott
et al. 2007).
Conclusion
In this study, captopril inhibited Dz-induced nephrotoxicity
and neurotoxicity that showed by amelioration of oxidative
stress markers in brain and kidney tissues and NO levels in
serum. In overall, we showed that captopril has beneficial
effects on kidney and brain oxidative damage following Dz
administration in rats. Therefore, captopril can be evaluated
for improving the pathological condition that oxidative stress
is involved.
Disclosure statement
No potential conflict of interest was reported by the authors.
Funding
This study was extracted from MSc thesis of Milad Vahidirad and sup-
ported by a grant from the Pharmaceutical Sciences Branch, Islamic Azad
University, Tehran, Iran.
References
Abdel-Daim, M.M., 2016. Synergistic protective role of ceftriaxone and
ascorbic acid against subacute diazinon-induced nephrotoxicity in rats.
Cytotechnology, 68 (2), 279–289.
Aebi, H., 1984. Catalase in vitro. Methods in Enzymology, 105, 121–126.
Ahangar, N., et al., 2017. Zinc deficiency and oxidative stress involved in
valproic acid induced hepatotoxicity: protection by zinc and selenium
supplementation. Biological Trace Element Research, 179 (1), 102–109.
Banerjee, A., et al., 2010. HIV proteins (gp120 and Tat) and methampheta-
mine in oxidative stress-induced damage in the brain: potential role of
the thiol antioxidant N-acetylcysteine amide. Free Radical Biology &
Medicine, 48 (10), 1388–1398.
Bedilli, H., et al., 2015. The effects of thymoquinone on nitric oxide and
supeoxide dismutase levels in a rat model of diazinon-induced brain
damage. Studies on Ethno-Medicine, 9 (2), 191–195.
Bengmark, S., 2006. Curcumin, an atoxic antioxidant and natural
NFkappaB, cyclooxygenase-2, lipooxygenase, and inducible nitric oxide
synthase inhibitor: a shield against acute and chronic diseases. Journal
of Parenteral and Enteral Nutrition, 30 (1), 45–51.
Beydilli, H., et al., 2015. Evaluation of the protective effect of silibinin
against diazinon induced hepatotoxicity and free-radical damage in
rat liver. Iranian Red Crescent Medical Journal, 17 (4), e25310.
Boroushaki, M.T., et al., 2013. Protective effect of pomegranate seed oil
against acute toxicity of diazinon in rat kidney. Iranian Journal of
Pharmaceutical Research, 12 (4), 821–827.
Boussabbeh, M., et al., 2016. Diazinon, an organophosphate pesticide,
induces oxidative stress and genotoxicity in cells deriving from large
intestine. Environmental Science and Pollution Research International, 23
(3), 2882–2889.


Colovi c, M.B., et al., 2015. In vitro evaluation of neurotoxicity potential
and oxidative stress responses of diazinon and its degradation prod-
ucts in rat brain synaptosomes. Toxicology Letters, 233 (1), 29–37.
Gao, X., et al., 2009. Huperzine A protects isolated rat brain mitochondria
against b-amyloid peptide. Free Radical Biology and Medicine, 46 (11),
1454–1462.
Ghazi-Khansari, M., Nasiri, G., and Honarjoo, M., 2005. Decreasing the oxi-
dant stress from paraquat in isolated perfused rat lung using captopril
and niacin. Archives of Toxicology, 79 (6), 341–345.
Giordano, G., et al., 2007. Organophosphorus insecticides chlorpyrifos and
diazinon and oxidative stress in neuronal cells in a genetic model of
glutathione deficiency. Toxicology and Applied Pharmacology, 219 (2),
181–189.
Hamad, M.H., et al., 2016. Acute poisoning in a child following topical
treatment of head lice (pediculosis capitis) with an organophosphate
pesticide. Sudanese Journal of Paediatrics, 16 (1), 63.
Heo, H., et al., 2008. Effects of banana, orange, and apple on oxidative
stress-induced neurotoxicity in PC12 cells. Journal of Food Science, 73
(2), H28–H32.
Hosseinimehr, S. and Karami, M., 2005. Chemoprotective effects of capto-
pril against cyclophosphamide-induced genotoxicity in mouse bone
marrow cells. Archives of Toxicology, 79 (8), 482–486.
Jafari, M., et al., 2012. The role of oxidative stress in diazinon-induced tis-
sues toxicity in Wistar and Norway rats. Toxicology Mechanisms and
Methods, 22 (8), 638–647.

Kalia, K., et al., 2007. Effects of combined administration of captopril and
DMSA on arsenite induced oxidative stress and blood and tissue
arsenic concentration in rats. Comparative Biochemistry and Physiology
Part C: Toxicology & Pharmacology, 144 (4), 372–379.
Kasperczyk, S., et al., 2014. Effect of N-acetylcysteine administration on
the expression and activities of antioxidant enzymes and the malon-
dialdehyde level in the blood of lead-exposed workers. Environmental
Toxicology and Pharmacology, 37 (2), 638–647.
Lasram, M.M., et al., 2014. Antioxidant and anti-inflammatory effects of
N-acetylcysteine against malathion-induced liver damages and immu-
notoxicity in rats. Life Sciences, 107 (1), 50–58.
Lukaszewicz-Hussain, A., 2010. Role of oxidative stress in organophos-
phate insecticide toxicity – Short review. Pesticide Biochemistry and
Physiology, 98 (2), 145–150.
Lushchak, V.I., 2012. Glutathione homeostasis and functions: potential tar-
gets for medical interventions. Journal of Amino Acids, 2012, 736837.
Naseh, M. and Vatanparast, J., 2014. Enhanced expression of hypothal-
amic nitric oxide synthase in rats developmentally exposed to organo-
phosphates. Brain Research, 1579, 10–19.
Noctor, G., Lelarge-Trouverie, C., and Mhamdi, A., 2015. The metabolo-
Downloaded by [University of New England] at 10:55 08 November 2017
mics of oxidative stress. Phytochemistry, 112, 33–53.
Oksay, T., et al., 2013. N-acetyl cysteine attenuates diazinon exposure-
induced oxidative stress in rat testis. Andrologia, 45 (3), 171–177.
Ortiz, M.S., et al., 2016. Effects of antioxidant N-acetylcysteine against
paraquat-induced oxidative stress in vital tissues of mice. International
Journal of Sciences, Basic and Applied Research, 26 (1), 26.
€ and Usta, D., 2007. Evaluation of oxidative stress responses
Oruç, E.O.
and neurotoxicity potential of diazinon in different tissues of Cyprinus
carpio. Environmental Toxicology and Pharmacology, 23 (1), 48–55.
Ott, M., et al., 2007. Mitochondria, oxidative stress and cell death.
Apoptosis: An International Journal on Programmed Cell Death, 12 (5),
913–922.
Parving, H.-H., et al., 2006. Prevalence and risk factors for microalbuminu-
ria in a referred cohort of type II diabetic patients: a global perspec-
tive. Kidney International, 69 (11), 2057–2063.
Peresleni, T., et al., 1996. Antisense oligodeoxynucleotides to inducible
NO synthase rescue epithelial cells from oxidative stress injury. The
American Journal of Physiology, 270 (6), F971–F977.
DRUG AND CHEMICAL TOXICOLOGY 7
Pizzurro, D.M., Dao, K., and Costa, L.G., 2014. Astrocytes protect against
diazinon-and diazoxon-induced inhibition of neurite outgrowth by
regulating neuronal glutathione. Toxicology, 318, 59–68.
Rashedinia, M., et al., 2016. Effect of exposure to diazinon on adult rat’s
brain. Toxicology and Industrial Health, 32 (4), 714–720.
Sadegh, C. and Schreck, R.P., 2003. The spectroscopic determination of
aqueous sulfite using Ellman’s reagent. MURJ, 8, 39–43.
Salem, N.A., Al Badawi, M.H., and Hussein, H.H., 2015. Protective role of
propolis on diazinon induced nephrotoxicity in adult male albino rats.
European Journal of Anatomy, 19 (4), 331–342.
Shahani, S., et al., 2016. Antioxidant and anti-inflammatory effects of
Nasturtium officinale involved in attenuation of gentamicin-induced
nephrotoxicity. Toxicology Mechanisms and Methods, 27 (2), 107–114.
Sharma, P., et al., 2012. Reactive oxygen species, oxidative damage, and
antioxidative defense mechanism in plants under stressful conditions.
Journal of Botany, 2012, 217037.
Standman, E. and Levine, R., 2000. Protein oxidation. Annals of the New
York Academy of Sciences, 899, 191–208.
Sulak, O., et al., 2005. Nephrotoxicity in rats induced by organophosphate
insecticide methidathion and ameliorating effects of vitamins E and C.
Pesticide Biochemistry and Physiology, 83 (1), 21–28.
Tsitsimpikou, C., et al., 2013. Histopathological lesions, oxidative
stress and genotoxic effects in liver and kidneys following long term
exposure of rabbits to diazinon and propoxur. Toxicology, 307,
109–114.
Tutanc, M., et al., 2012. Effects of erdosteine on cyclosporin-A-induced
nephrotoxicity. Human & Experimental Toxicology, 31 (6), 565–573.
Uhlenius, N., et al., 1999. Renoprotective effects of captopril in hyperten-
sion induced by nitric oxide synthase inhibition in experimental neph-
ritis. Nephron, 81 (2), 221–229.
Yavuz, D., et al., 2003. Effects of captopril and losartan on lipid peroxida-
tion, protein oxidation and nitric oxide release in diabetic rat kidney.
Prostaglandins, Leukotrienes and Essential Fatty Acids, 69 (4), 223–227.
Zhang, F., et al., 2008. In vitro effect of manganese chloride exposure on
energy metabolism and oxidative damage of mitochondria isolated
from rat brain. Environmental Toxicology and Pharmacology, 26 (2),
232–236.