Article

Toxicology and Industrial Health

2015, Vol. 31(9) 841–851
Hesperidin protects brain and sciatic nerve © The Author(s) 2013
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tissues against cisplatin-induced oxidative, DOI: 10.1177/0748233713483192
tih.sagepub.com
histological and electromyographical side
effects in rats

Suat Kamisli1, Osman Ciftci2, Kursat Kaya3, Asli Cetin4,

Ozden Kamisli1 and Cemal Ozcan1

Abstract
In the present study, the beneficial effect of hesperidin (HP), a citrus flavonoid, on cisplatin (CP)-induced
neurotoxicity was investigated. A total of 28 rats were equally divided into four groups; the first group was kept
as control. In the second and third groups, CP and HP were given at the doses of 7 and 50 mg/kg/day,
respectively. In the fourth group, CP and HP were given together at the same doses. The results indicated that
although CP caused significant induction of lipid peroxidations and reduction in the antioxidant defense system
potency in the brain and sciatic nerve, HP prevented these effects of CP. Besides, CP led to histopathological
damage, mainly apoptosis, as well as electromyographical (EMG) changes in sciatic nerve. On the other hand,
HP treatment reversed histopathological and EMG effects of CP. In conclusion, CP had severe dose-limiting
neurotoxic effects and these effects of CP can be prevented by HP treatment. Thus, it appears that coadminis-
tration of HP with CP may be a useful approach to attenuate the negative effects of CP on the nervous system.

Keywords
Cisplatin, hesperidin, neurotoxic effect, oxidative damage, electromyography

Introduction (EMG) values (reduced amplitude and induced

latency), painful paresthesias of the extremities and
Cisplatin (cis-diamminedichloroplatinum(II); CP) is
numbness, and loss of vibration sense and ataxia. In
an effective chemotherapeutic agent used for the
addition, both short- and long-term administration of
treatment of many types of cancers including testis,
CP cause a significant oxidative stress by forming
lung, colorectal, ovarian, breast, and bladder (Carozzi
reactive oxygen species or free radicals with a conco-
et al., 2010). The adverse effects including reproduc-
mitant increase in lipid peroxidation and decline in
tive nephrotoxicity and neurotoxicity limit the clinical
usage of CP. Neurotoxicity is one of the most impor-
tant adverse effects of CP and is caused due to dose-
1
limiting CP (Orhan et al., 2004). The neurotoxic Department of Neurology, Faculty of Medicine, University of
mechanism of CP may be due to the fact that platinum Inonu, Malatya, Turkey
2
Department of Pharmaceutical Toxicology, Faculty of Pharmacy,
accumulates and leads to oxidative damage and apop-
University of Inonu, Malatya, Turkey
totic cell death in the nervous system (Carozzi et al., 3
Department of Medical Biochemistry, Graduate Institute of
2010). Oxidative stress caused by reactive oxygen Health Sciences, University of Inonu, Malatya, Turkey
4
species can be mainly responsible for peripheral and Department of Histology and Embryology, Faculty Medicine,
central neurotoxicity of CP (Rosenberg, 1978). Sev- University of Inonu, Malatya, Turkey
eral in vivo and in vitro studies (Barabas et al., 2008;
Corresponding author:
Krarup-Hansen et al., 1993) have demonstrated that Osman Ciftci, Department of Pharmaceutical Toxicology, Faculty
the side effects of CP on nervous system are character- of Pharmacy, University of Inonu, Malatya 44280, Turkey.
ized by significant changes in the electromyographical Email: osmciftci@gmail.com

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842 Toxicology and Industrial Health 31(9)

antioxidant enzyme activity in tissues (Beytur et al., were housed in sterilized polypropylene rat cages, in
2012; Hartmann et al., 1999; Yüce et al., 2007). Some a 12-h light–dark cycle, at an ambient temperature
experimental studies (Olas and Wachowicz, 2004; of 21 C. Diet and water for them were given ad libi-
Orhan et al., 2004; Rodriguez-Menendez et al., 2008; tum. Experiments were performed based on animal
Tuncer et al., 2010) showed that the use of antioxidant ethics guidelines of the Institutional Animals Ethics
therapies such as resveratrol, erythropoietin, melatho- Committee.
nin, alpha-lipoic acid, and valproate reduces CP neuro- Rats were randomly divided into four equal groups
toxicity in animal model. However, there is no study as (n ¼ 7 in each group). CP was intraperitoneally (i.p.)
to whether or not the hesperidin (HP) treatment can administered at the dose of 7 mg/kg with a single
prevent CP neurotoxicity. injection. HP was given at the doses of 50 mg/kg/day
HP is a bioflavonoid, which is believed to play a for 14 consecutive days by gavages. Group 1 (control)
role in plant defense, and is found abundantly in served as negative control and was given isotonic
citrus species such as lemon and orange. Various saline (i.p.) and corn oil (orally) as vehicles. In group
previous studies (Choi and Ahn, 2008; Menze 2 (CP group), CP was given as a single injection and
et al., 2012) revealed that HP exhibits pharmacologi- then corn oil was given for 14 days. Rats in group 3
cal activities including antioxidant, anticarcino- (HP group) were treated with HP for 14 days without
genic, antihypertensive, and anti-inflammatory CP. In group 4, the rats were treated with CP and HP
activities but none of which has been confirmed as (i.e. CP þ HP group) together. Tissue samples were
applicable to humans. Shagirta and Pari (2011) collected on day 14 of CP treatment. The animals
determined that HP treatment protects testicular were killed under ether anesthesia and tissues (brain
functions against cadmium toxicity because of anti- and sciatic nerve) were immediately removed and
oxidant properties. It is therefore believed that HP dissected over ice-cold glass. Tissue samples were
is a powerful radical scavenger that promotes cellu- stored at 86 C deep freezer until analysis.
lar antioxidant defense system and can also traverse
the brain–blood barrier (Shagirtha and Pari, 2011;
Youdim et al., 2003). Thus, HP can prevent neurode- Biochemical assay
generation caused by toxic agents and can promote The homogenization of tissues was carried out in
brain functions (Hwang et al., 2012). Teflon glass homogenizer with 150 mM potassium
Based on this background, we hypothesized that chloride (pH 7.4) to obtain 1:10 (weight in volume)
HP may prevent CP-induced neurotoxicity due to its dilution of the whole homogenate. The homogenates
intrinsic biochemical and antioxidant properties. To were centrifuged at 18,000 g (4 C) for 30 min to
this end, oxidative status, histopathological, immuno- determine thiobarbituric acid reactive substances
histochemical, and EMG changes in the brain and (TBARS), reduced glutathione (GSH) levels, and
sciatic nerve of rats were investigated. catalase (CAT) activities and at 25,000 g for 50 min
to determine glutathione peroxidase (GPx) and
copper-/zinc-superoxide dismutase (SOD) activities.
Materials and methods The levels of homogenized tissue TBARS, as an
Chemicals index of lipid peroxidation, were determined by thio-
barbituric acid (TBA) reaction using the method of
CP (10 mg/10 mL, code 1876A) was obtained from
Yagi (1998). The product was evaluated spectropho-
Faulding Pharmaceuticals Plc (Warwickshire, UK).
tometrically at 532 nm and the results are expressed
HP was given from Molecula Limited (Gillingham,
in nanomole per gram tissue. The GSH content of the
UK) All other chemicals were purchased from Sigma
brain and nerve homogenate was measured at 412 nm
Chemical Co. (St Louis, Missouri, USA) and were of
using the method of Sedlak and Lindsay (1968). The
analytical grade or of the highest grade available.
GSH level was expressed in nanomole per milliliter.
SOD activity was measured by the inhibition of nitro-
Animals and treatment blue tetrazolium (NBT) reduction due to O2 gener-
A total of 28 healthy adult male Spraque Dawley rats ated by the xanthine/xanthine oxidase system (Sun
(aged between 2 and 3 months and weighing 250–300 et al., 1988). One unit of SOD activity was defined
g) were obtained from the Experimental Animal Insti- as the amount of protein causing 50% inhibition of the
tute, Malatya, Turkey, for this experiment. Animals NBT reduction rate. The product was evaluated

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Kamisli et al. 843

spectrophotometrically at 560 nm. Results are EMG assay

expressed in international unit per milligram of protein. On day 14 of drug administration, the EMG changes
CAT activity of the tissues was determined according were performed under general anesthesia and were car-
to the method of Aebi (1974). The enzymatic decom- ried out on the right sciatic nerve with a Dantec Key-
position of hydrogen peroxide (H2O2) was followed point electromyography (code number, 9020M0203
directly by the decrease in the absorbance at 240 nm. and serial number, 1562, Medtronic, Skovlunde, Den-
The difference in absorbance per unit time was used mark). Neurophysiological analysis was performed
as a measure of CAT activity. The enzyme activities according to the method of Kalender et al. (2009).
are given in k (katalase activity) per milligram of pro- Briefly, for the stimulations, the sciatic nerve was per-
tein. GPx activity was measured by the method of cutaneously stimulated with supramaximal stimulus
Paglia and Valentina (1967). In the presence of glu- for the determination of compound muscle action
tathione reductase and nicotinamide adenine dinu- potentials (CMAPs) with monopolar needle electrodes.
cleotide phosphate (NADPH), the oxidized Electrical stimulation was square pulse with a fre-
glutathione is immediately converted to the reduced quency of 1 Hz and duration of 0.2 ms. Recorded sig-
form with a concomitant oxidation of NADPH to nals were amplified by an alternating current-coupled
nicotinamide adenine dinucleotide phosphate. The preamplifier with filters at 1 Hz and 10 KHz. CMAP
decrease in absorbance at 340 nm was measured. GPx records were obtained from distal gastrocnemius mus-
activity is expressed as international unit per milli- cle with needle electrodes. The common reference
gram of protein. Tissue protein content was deter- (ground electrode) was placed on the tail. The CMAP
mined according to the method developed by Lowry amplitudes were measured from peak to peak and the
et al. (1951) using bovine serum albumin as standard. latencies were measured from the stimulus artifact to
the first deflection from baseline.
Histological evaluation
For light microscopic evaluation, brain and nerve
Statistical analysis
samples were fixed in 10% formalin and were All values were presented as mean + SEM. Significant
embedded in paraffin. Paraffin-embedded specimens differences (p < 0.05) are given in tables. A computer
were cut into 5-mm-thick sections, mounted on slides, program SPSS 18.0 (SPSS Inc., Chicago, Illinois, USA)
and stained with hematoxylin–eosin (H&E) stain. Tis- was used for statistical analysis. For the comparison
sue samples were examined using a Leica DFC280 of biochemical and EMG parameters, statistical analy-
light microscope and a Leica Q Win Image Analysis ses were performed using one-way analysis of variance
system (Leica Micros Imaging Solutions Ltd, Cam- and post hoc Duncan’s significant difference test.
bridge, UK).
For immunohistochemical analysis, thick sections Results
were mounted on polylysine-coated slides. After
rehydrating, samples were transferred to citrate buffer Biochemical results
(pH 7.6) and heated in a microwave oven for 20 min. The brain SOD, GPx, CAT, GSH, and TBARS values
After cooling for 20 min at room temperature, the sec- are given in Table 1. The results demonstrated that CP
tions were washed using phosphate-buffered saline caused a significant increase in TBARS levels and
(PBS). Then sections were kept in 0.3% H2O2 for 7 decrease in GSH levels and antioxidant enzyme activi-
min and afterward washed with PBS. Sections were ties (CAT, SOD, and GPx) in brain tissues compared
incubated with primary rabbit polyclonal caspase-3 with other groups. Additionally, HP administration
antibody (Abcam, Cambridge, MA, USA, Ab4051) lowered TBARS and elevated GSH levels compared
for 2 h. They were then rinsed using PBS and incu- with control values. Additionally, HP treatments
bated with biotinylated goat anti-polyvalent for 10 together with CP led to significant decline in elevated
min and streptavidin peroxidase for 10 min at room TBARS levels and induction in antioxidant enzyme
temperature. Staining was completed with chromogen activity and GSH levels close to the values of the control
þ substrate for 15 min, and then the slides were coun- group.
terstained with Mayer’s hematoxylin for 1 min, rinsed The levels of TBARS, SOD, GPx, CAT, and GSH
in tap water, and dehydrated. The caspase-3 kit was in sciatic nerve tissue are given in Table 2. TBARS
used according to the manufacturer’s instructions. levels were significantly increased and SOD, GPx,

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844 Toxicology and Industrial Health 31(9)

Table 1. Changes in SOD, CAT, and GPx activities and GSH and TBARS levels in brain tissue of rats administered with CP
and HP (mean + SEM; n ¼ 7).a
TBARS (nmol/g tissue) GSH (nmol/mL) SOD (U/mg protein) CAT (k/mg protein) GPx (U/mg protein)
Control 8.60 + 0.28b 90.4 + 1.49b 34.2 + 2.46b 0.024 + 0.001b 2.39 + 0.073b
CP 11.88 + 0.72c 71.5 + 2.25c 18.2 + 1.48c 0.014 + 0.0003c 1.23 + 0.090c
HP 6.62 + 0.43d 98.8 + 3.31d 34.4 + 1.74b 0.027 + 0.0009b 2.41 + 0.081b
CP þ HP 9.62 + 0.67b 86.3 + 1.16b 26.0 + 1.04d 0.018 + 0.0005d 1.93 + 0.112d
SOD: superoxide dismutase; CAT: catalase; GPx: glutathione peroxidase; GSH: glutathione; TBARS: thiobarbituric acid reactive sub-
stances; CP: cisplatin; HP: hesperidin.
a
Means bearing different superscripts within same column were significantly different (p  0.05).

Table 2. Changes in SOD, CAT, and GPx activities and GSH and TBARS levels in nerve tissue of rats administered with
CP and HP (mean + SEM; n ¼ 7).a
TBARS (nmol/g tissue) GSH (nmol/mL) SOD (U/mg protein) CAT (k/mg protein) GPx (U/mg protein)
Control 4.32 + 0.33b 225.4 + 11.4b 3.86 + 0.16b 0.023 + 0.0009b 1.95 + 0.121b
CP 7.36 + 0.51c 133.3 + 2.8c 1.88 + 0.27c 0.014 + 0.0003c 1.12 + 0.087c
HP 4.03 + 0.23b 229.0 + 8.5b 4.01 + 0.30b 0.024 + 0.001b 2.05 + 0.130b
CP þ HP 4.83 + 0.26b 170.4 + 7.9d 3.03 + 0.23d 0.017 + 0.0007b 1.83 + 0.115b
SOD: superoxide dismutase; CAT: catalase; GPx: glutathione peroxidase; GSH: glutathione; TBARS: thiobarbituric acid reactive sub-
stances; CP: cisplatin; HP: hesperidin.
a
Means bearing different superscripts within same column were significantly different (p  0.05).

CAT, and GSH levels were significantly decreased in However, CP led to deeply stained, shrunken, and
CP group compared with control and other groups. variously shaped Purkinje cells with pyknotic nuclei
However, elevated TBARS levels and lowered SOD, (Figure 2(b)), and HP prevented these changes when
GPx, CAT, and GSH levels were significantly combined with CP (Figure 2(c)).
reversed to the value of the control group in rats In immunohistochemical analysis, caspase-3-stain-
treated with HP þ CP (Table 2). ed cells were not observed in the cerebral cortex of
control (Figure 3(a)) and HP (Figure 3(d)) groups.
On the other hand, the percentage of caspase-3-
Histopathological and immunohistochemical positive cells was significantly higher in the CP group
results than other groups (Figure 3(b)). Besides, the density
of caspase-3-stained cells was minimal (Figure 3(c))
Histological findings indicated that the brain tissue
in CP þ HP group. The sciatic nerve was evaluated
had a normal histological appearance in control and
as transverse sections, and the sciatic nerve degenera-
HP groups. On the other hand, in CP (Figure 1(a)) and
tion (axonal degeneration and demyelination) was
CP þ HP (Figure 1(b)) groups, the cell infiltration in
determined in CP (Figure 4(b)) and CP þ HP groups
cerebral cortex was observed compared with that of
(Figure 4(c)). However, these findings were decreased
control (Figure 1(c)) and HP (Figure 1(d)) groups.
in CP þ HP groups compared with CP group. Besides,
However, in CP þ HP group (Figure 1(b)), the cell
in control (Figure 4(a)) and HP (Figure 4(d)) groups,
infiltration was significantly decreased in comparison
sciatic nerve showed normal histological appearance.
with CP group (Figure 1(a)). Also, shrinkage of the
cytoplasm and extensively dark pyknotic nuclei in
neurons of the cerebral cortex tissue were seen in the EMG results
CP group (Figure 1(e)). These abnormalities were sig- The latency and amplitude values in sciatic nerve of
nificantly alleviated when HP has been given together rats exposed to CP and HP are given in Table 3. The
with CP (Figure 1(f); H&E; 40). The cerebellar cor- latency time significantly prolonged and amplitude
tex showed normal histological appearance, and Pur- value significantly declined with CP treatment.
kinje cells showed no histological abnormalities in However, HP administration increased the lowered
control (Figure 2(a)) and HP (Figure 2(d)) groups. amplitude value and decreased the elevated latency

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Kamisli et al. 845

Figure 1. In CP group (a), intensive cell infiltration (arrows) was detected. This infiltration (arrows) was decreased in
CP þ HP group (b); H&E, 20. In control (c) and HP (d) groups, neurons of cerebral cortex showed normal histological
appearance. In CP group (e), shrinkage cytoplasm and extensively dark pyknotic nuclei in neurons (arrows) of the cerebral
cortex tissues were seen. These findings (arrows) were decreased in CP þ HP groups (f); H&E, 40. CP: cisplatin; HP:
hesperidin; H&E: hematoxylin–eosin.

back to the control value. These results also indicated

that although CP treatment negatively affected CMAPs Discussion
(decreased amplitude and increased latency), HP treat- CP treatment can cause neurological side effects in
ment completely brought CMAP values closer to cancer patients and this situation can be limiting the
control value. clinical dosage of CP. For that, the prevention of side

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846 Toxicology and Industrial Health 31(9)

Figure 2. In control (a) and HP (d) groups, cerebellar cortex had showed normal histological appearance. In CP group (b),
deeply stained Purkinje cells with pyknotic nuclei (arrows) were seen. In CP þ HP group (c) Purkinje cell degeneration
(arrows) was decreased; H&E, 40. CP: cisplatin; HP: hesperidin; H&E: hematoxylin–eosin.

effects of CP is very important in terms of treatment 2010; Hwang et al., 2012) indicated that CP led to
protocol, benefits in life quality, and widening the dose neurotoxic effects in human and animals via induction
limits. In the present study, it was determined that CP- of lipid peroxidations. Our study showed that CP pro-
induced neurotoxic effects via increased oxidative duced a significant increase in the TBARS levels; as a
stress, histopathological defects, and EMG changes result, it induced lipid peroxidations in the brain and
also decreased antioxidant status in rats. However, sciatic nerve. Also the enzyme activity of antioxidants
HP treatment can reverse toxic effects of CP on the (SOD, CAT, and GPx) was suppressed and the GSH
nervous system when used together with CP. levels were declined with CP treatment. Therefore,
CP clearly led to an imbalance of oxidative system
and caused oxidative damage in central and peripheral
Oxidative evaluations nervous system. Carozzi et al. (2010) indicated that
Oxidative stress, a condition of an imbalance between oxidative stress has a significant role in the neurologi-
the free radicals and antioxidant defense system, is an cal toxicity of platinum complex and may be the pri-
important factor in the pathogenesis of neurological mary effect mechanism of CP on the nervous system.
disorders due to the fact that the nervous system has Rybak et al. (2000) indicated that CP caused 165% ele-
high content of polyunsaturated membrane lipids vation in malondialdehyde (MDA) levels, nearly 50%
(Acar et al., 2012). Recent studies (Carozzi et al., depletion of GSH, and about 50% reduction in the

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Kamisli et al. 847

Figure 3. Immunohistochemical expression of caspase-3 in the cortex of control and experimental groups of rats. In con-
trol (a) and HP groups (d), there were no caspase-3-stained cells. Caspase-3-stained cells (arrows) increased in CP group
(b). The number of positive-stained cells (arrows) decreased in CP þHP group (c); caspase-3, 40. CP: cisplatin; HP:
hesperidin.

activities of SOD, GPx, and GSH reductase, while oxidative stress (Choi and Ahn, 2008; Hwang and
CAT activity was reduced to 70% of the control values Yen, 2008; Hwang et al., 2012). Indeed, the present
in rats. Also Qu et al. (2012) reported that MDA study demonstrated that HP therapy could signifi-
levels significantly increased; however, antioxidant cantly prevent oxidative stress through returning the
parameters decreased upon CP treatment in the nervous elevated TBARS levels and declined antioxidant
system. These studies (Rybak et al., 2000; Qu et al., enzyme activity and GSH levels back to the normal
2012) showed that CP and other platinum complexes in the brain and sciatic nerve when given together
produced oxidative changes in the nervous system and with CP. Choi and Ahn (2008) investigated neuropro-
these results agree with our findings. Therefore, our tective effects of HP in mice and they suggested that
results clearly demonstrate that CP induces oxidative chronic HP treatment has a neuroprotective effect by
damage in the brain and sciatic nerve of the rats. inhibiting the oxidative damage (decline of TBARS),
Several researchers suggested that citrus flavo- together with activating the antioxidant defense
noids such as HP have strong antioxidant activity and system. In the same study, TBARS levels were
they hypothesized that the antioxidant action of flavo- declined by 21.36%, while SOD, GPx, and GSH lev-
noids may reduce multi-organ toxicity caused by els were increased by 20.58%, 15.15%, and 21.83%,

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848 Toxicology and Industrial Health 31(9)

Figure 4. We compared sciatic nerve transverse sections in all the groups. The control (a) and HP (d) groups showed
normal histological appearance. In CP group (b), nerve degeneration was prominent. This degeneration was decreased in
CP þ HP group (c); H&E, 100. CP: cisplatin; HP: hesperidin; H&E: hematoxylin–eosin.

Table 3. The CMAP values for CP and HP groups (n ¼ 7, oxicity in rats and indicated that HP significantly pre-
mean + SEM).a vented neurotoxic effects via its antioxidant proper-
ties. All the results from previous studies agree with
Group Latency (ms) Amplitude (mV)
our results. Finally, HP treatment can be beneficial for
Control 1.32 + 0.051b 13.11 + 0.73b the neurotoxic effects of CP when given together, and
CP 2.31 + 0.112c 10.22 + 0.57c neuroprotection may be due to its radical scavenging,
HP 1.20 + 0.043b 14.70 + 0.61b antioxidant, and anti-inflammatory activities.
CP þ HP 1.25 + 0.048b 13.34 + 0.61b
CMAP: compound muscle action potential; CP: cisplatin; HP: Histological changes
hesperidin.
a
Means bearing different superscripts within same column were In the present study, it was demonstrated that CP
significantly different (p  0.05). significantly produced histopathological alterations in
the brain and sciatic nerve tissues. Primarily, hemor-
respectively, with HP treatment in mice. Besides, rhage in arachnoid layer, cell infiltration in cerebral
Menze et al. (2012) investigated the potential effects cortex, shrinkage in the cytoplasm, and extensively
of HP against 3-nitropropionic acid-induced neurot dark pyknotic nuclei in neurons of the cerebral cortex

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Kamisli et al. 849

were observed in the brain tissue after CP treatment. preventing CP effects on peripheral neuropathy, many
Additionally, apoptosis was clearly seen in neurons antioxidant agents were tested such as melathonin,
of the cerebral cortex upon immunohistochemical eva- alpha lipoic acid, and valproate (e.g., Orhan et al.,
luation, and these data were supported by caspase-3 2004; Rodriguez-Menendez et al., 2008; Tuncer et al.,
staining. Demyelination and axonal degeneration 2010) and some of these were found to be efficient.
occurred in sciatic nerve after CP administration. How- When given together with CP, HP prevented side effects
ever, histopathological damages formed were signifi- of CP in terms of peripheral neuropathy via a significant
cantly reversed with HP-combined treatment. Results decline in elevated latency value and induction in lower
from previous studies (Abou-Elghait et al., 2010; amplitude value. These results suggested that HP treat-
Kirchmair et al., 2005) are in agreement with our find- ment can fully reverse peripheral neuropathy induced
ings, and they determined that CP treatment increased by CP with changing latencies and amplitude values.
histopathological damage in brain and sciatic nerve tis-
sue. Menze et al. (2012) also reported similar findings Conclusion
that HP treatment can be beneficial for the protection
In conclusion, the present study revealed the toxic
of the nervous system against 3-nitropropionic acid-
effects of CP on central and peripheral nervous
induced neurotoxicity in rats. It was considered that the
system with oxidative stress, histopathological
histopathological effects may be attributed to the
damage, and EMG alterations. Also, the use of HP
imbalance between oxidant and antioxidant status
in combination with CP minimized its toxicity, which
induced by CP. Moreover, this situation may contribute
was evident from decreasing TBARS levels, histolo-
to severe neurological disorders such as peripheral neu-
gical changes in tissue, EMG alterations, and increas-
ropathy. For this reason, a decrease in elevated oxida-
ing antioxidant enzyme activities (SOD, CAT, and
tive stress in the nervous system as a result of HP
GPx) and GSH levels. The beneficial effects of HP
treatment is of importance in CP neurotoxicity.
against CP-induced neurological damage may be due
to its antioxidant, anti-inflammatory, and free radical-
EMG alterations scavenging properties. Therefore, it appears that HP, a
citrus flavonoid, can prevent and protect against
Peripheral neurotoxicity, determined by CMAPs
neurological damage caused by CP in rats.
latency and CMAPs amplitude with electromyography,
is the main toxic effect of CP. First, we showed that CP Conflict of interest
administration significantly prolonged the CMAPs The authors declared no conflict of interest.
latency. Also the mean amplitude of CMAPs, depend-
ing on the number of axons that conduct impulses Funding
from the stimulus point to the muscle, the number of This research received no specific grant from any funding
functioning motor end-plates, and the muscle volume agency in the public, commercial, or not-for-profit sectors.
(Korte et al., 2011; Stålberg and Erdem, 2002) were sig-
nificantly declined with CP treatment. Because CMAPs References
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