Effect of lamotrigine, oxcarbazepine and topiramate on cognitive functions

and oxidative stress in PTZ-kindled mice

Nidhi Bharal Agarwal
a,c
, Nitin Kumar Agarwal
b
, Pramod Kumari Mediratta
a,
*, Krishna Kishore Sharma
a
a
Department of Pharmacology, University College of Medical Sciences (University of Delhi) & GTB Hospital, New Delhi 110095, India
b
Corpotate Quality Assurance GP5, Ranbaxy Research Laboratories, Gurgaon 122001, Haryana, India
c
Department of Clinical Research, Faculty of Allied Health Sciences, Jamia Hamdard (Hamdard University), New Delhi 110062, India
1. Introduction
Epilepsy is a common neurological disorder that affects
approximately 50 million people worldwide.
1
Epilepsy and
antiepileptic drugs (AEDs) therapy have deleterious effects on
cognition in patients receiving them. A large number of the
epileptic patients suffer frommemory decits, learning disabilities
and behavioural problems.
68
Several factors viz.: biological
(underlying neuropathology, seizure type and aetiology, age of
onset, frequency, severity and duration of seizures, structural
cerebral damage), psychological problems, treatment with AEDs
and by surgery have been attributed to cause neurocognitive
impairment.
2
It has been noticed that AEDs can also brings about
untoward effect on cognition and behaviour.
35
It has been
reported that the neurocognitive burden of epilepsy may even start
before birth through in utero exposure to AEDs.
1315
At therapeutic
doses AED treatment disturbs memory formation, retention and
orientation. These adverse effects were especially prominent in
patients receiving polytherapy.
35
Among the established antiepi-
leptic drugs barbiturates and benzodiazepines have demonstrated
detrimental effects on cognition leading to decreased arousal and
decits in cognitive performance.
9,10
Phenobarbital has more
cognitive and behavioural toxicity as compared to carbamazepine,
sodium valproate, clonazepam and phenytoin which cause mild
cognitive impairment.
17
However, some of the newer antiepileptic
drugs have been reported to have favourable cognitive and
behavioural prole such as vigabatrine, gabapentin, levetiracetam,
tiagabine.
17
Among the newer drugs topiramate administration to
epileptic patients has brought about long term harmful effects on
the cognitive functions and verbal uency.
1821
It has been noticed
that because of cognitive impairment the drug needs to be
discontinued even when it is found to be improving seizure
frequency.
18
However there are some clinical and preclinical
reports available which suggest that topiramate treatment has no
negative effect on cognitive functions.
22,23
Although many
researchers have suggested that lamotrigine is a safe drug
2426
but there are certain indications which show that it might impair
Seizure 20 (2011) 257262
A R T I C L E I N F O
Article history:
Received 1 June 2010
Received in revised form 7 December 2010
Accepted 14 December 2010
Keywords:
Epilepsy
Kindling
Cognition
Oxidative stress
Topiramate
A B S T R A C T
Cognitive impairment is frequently observed in epileptic patients. It has been seen that not only epilepsy
but antiepileptic drugs also impair cognitive functions. The present study was undertaken to assess the
effect of three anticonvulsants viz. lamotrigine (5 mg/kg, p.o.), oxcarbazepine (15 mg/kg, p.o.) and
topiramate (10 mg/kg, p.o.) on cognitive function and oxidative stress during pentylenetetrazole (PTZ)-
kindling in mice. Kindling was induced by the administration of PTZ (25 mg/kg, i.p.) on every alternate
day till 5 weeks. Cognition was assessed after the development of kindling. Elevated plus maze (EPM)
and passive avoidance response (PAR) tests were carried out after 24 h and 48 h of the last PTZ
administration. After completion of behavioural tests malondialdehyde (MDA), glutathione levels,
superoxide dismutase and catalase activity were measured as an indicator of oxidative stress. The results
of the present study indicate that topiramate (10 mg/kg) administration to kindled animals increased
transfer latency and decreased step-down latency in EPM and PAR tests, respectively. However,
lamotrigine and oxcarbazepine did not alter the two parameters. Topiramate administration to kindled
as well as non-kindled animals has shown increase in MDA and decrease in glutathione levels.
Lamotrigine and oxcarbazepine did not show signicant alteration in oxidative stress parameters. To
conclude, long term administration of topiramate impairs cognitive functions during experimental
epilepsy while lamotrigine and oxcarbazepine are safer.
2010 British Epilepsy Association. Published by Elsevier Ltd. All rights reserved.
* Corresponding author at: Department of Pharmacology, University College of
Medical Sciences (University of Delhi) & GTB Hospital, New Delhi 110095, India.
Tel.: +91 11 22592972x75; fax: +91 11 22590495; mobile: +91 9810524879.
E-mail addresses: nidhi.bharal@gmail.com (N.B. Agarwal),
nitin.agarwal@ranbaxy.com (N.K. Agarwal), drpramod_k@yahoo.com
(P.K. Mediratta), drkksharma2000@yahoo.com (K.K. Sharma).
Contents lists available at ScienceDirect
Seizure
j our nal homepage: www. el sevi er . com/ l ocat e/ ysei z
1059-1311/$ see front matter 2010 British Epilepsy Association. Published by Elsevier Ltd. All rights reserved.
doi:10.1016/j.seizure.2010.12.006
memory.
16
Studies meant for assessing the effect of oxcarbazepine
on cognitive functions in healthy human volunteers indicate a
stimulant effect on memory and psychomotor functioning.
55
Oxcarbazepine treatment for over 6 months in children and
adolescents with newly diagnosed partial seizures indicated that it
does not affect cognition and intelligence as compared to
carbamazepine and valproate.
56
Interestingly, a cross-sectional
study indicated superiority of oxcarbazepine and lamotrigine
compared to topiramate, valproic acid, carbamazepine and lithium
in case of neurocognitive effects in patients suffering from mood
disorders.
57
Thus it would be interesting to assess the comparative
effects of three antiepileptic drugs viz. lamotrigine, oxcarbazepine
and topiramate on cognitive functions. This comparison would
further help in the decision to choose one drug over another.
A number of experimental and clinical reports suggest the
involvement of oxidative stress in pathophysiology of epilepsy.
27
The increased free radicals result in membrane lipid peroxidation
and decreased glutathione concentrations in the epileptic focus.
28
Further, the involvement of freeradicals inseizures is alsosupported
by reports which indicate that exogenously administered antioxi-
dant protects the brain against seizures.
29
Growing evidence
indicates that long term antiepileptic drug treatment leads to
increase in oxidative stress which is similar to that observed during
epileptogenesis. For example, valproic acid has been found to
increase lipidperoxidationinpatients receiving it.
30
Contrary to this
observation some anti-epileptic agents like phenytoin have been
shown to decrease oxidative stress demonstrated by increase in
glutathione reductase activity in patients receiving it.
31
Prolonged oxidative damage to various brain sites have been
suggested to be the more likely cause responsible for long term
complications, morphological abnormalities and cognition im-
pairment in neurodegenerative disorders.
11,12
Further, increased
free radicals lead to neuronal degeneration through increased lipid
peroxidation, decreased glutathione levels and decrease in super-
oxide dismutase and catalase activites.
32
It is not clear whether
antiepileptic drugs exert derangement of cognitivefunctionthrough
oxidative stress in spite of showing an antiepileptic effect.
In perspective of the above citations, the present study was
planned to assess the per se effect of newer antiepileptic drugs viz.
lamotrigine, topiramate and oxcarbazepine on cognitive functions
in kindled and non-kindled mice and to determine if oxidative
stress was involved in the derangement of cognition.
2. Methods
2.1. Animals
Male albino Swiss strain mice weighing 2530 g were procured
from Central Animal House of University College of Medical
Sciences (University of Delhi) and Guru Tegh Bahadur Hospital,
Delhi, India. The animals were housed in propylene cages in groups
of 68 mice per cage and kept under controlled environmental
condition (temperature: 22 2 8C), humidity: 5055%, natural light/
day cycle). The mice received a standard pellet diet (Gold Mohar
Lipton India Ltd., Kolkata) and water ad libitum. The animals were
brought to the laboratory at least two hours before the experiments
for acclimatization to the new premises. All the experiments were
performed during the light phase between 9:00 AM and 4.00 PM. The
work protocol was approved by Institutional Animal Ethics Commit-
tee and care of the animals was taken as per CPCSEA Guidelines for
laboratory animal facilities.
2.2. Drugs and dosing schedule
Lamotrigine and oxcarbazepine were gift sample from Sun
Pharma, Ahmedabad. Topiramate (Topamax) tablets were pur-
chased from market. Other (inactive) ingredients of Topamax are:
lactose monohydrate, pregelatinized starch, micro crystalline
cellulose, sodium starch glycolate, magnesium stearate, puried
water, carnauba wax, hypromellose, titanium dioxide, polyethyl-
ene glycol, synthetic iron oxide and polysorbate 80. Lamotrigine,
oxcarbazepine and topiramate tablets (after crushing) were
dissolved in 1% CMC (carboxymethyl cellulose) solution. All Drugs
were administered orally in a dose of 10 ml/kg. Pentylenetetrazole
(Sigma Aldrich, USA) was dissolved in double distilled water and
administered in a dose of 25 mg/kg, i.p. on every alternate day to
induce kindling. Drug treatment was given till ve weeks. The
doses of drugs were standardized on the basis of the pilot studies
conducted in the laboratory. Lamotrigine in dose of 5 mg/kg, p.o.,
oxcarbazepine in a dose of 15 mg/kg, p.o. and topiramate in a dose
of 10 mg/kg, p.o. showed signicant anticonvulsant effect in
maximal electroshock seizure test (signicant reduction of tonic
extensor phase) and delayed PTZ-induced kindling in mice.
33
2.3. Pentylenetetrazole induced kindling
For PTZ kindling, a subconvulsant dose of PTZ 25 mg/kg body
weight was injected intraperitoneally on every second day (i.e. Day
1, Day 3, Day 5. . .). The PTZ injections were stopped when the
animals showed adequate kindling, i.e. seizure score of 5 on three
consecutive injections. The rst incidence of seizure with score ve
was observed between Day 27 and Day 31. Thus, in no case did the
PTZ schedule exceed Day 35. In the groups treated with
antiepileptic drugs, the PTZ doses were tested up to Day 35 or
up to seizure of score 5 on three consecutive injections, whichever
was earlier. After each PTZ injection, the convulsive behaviour was
observed for 30 min. The resultant seizures were scored as follows:
Stage 0 (no response); Stage 1 (hyperactivity, restlessness and
vibrissae twitching); Stage 2 (head nodding, head clonus and
myoclonic jerks); Stage 3 (unilateral or bilateral limb clonus);
Stage 4 (forelimb clonic seizures); Stage 5 (generalized clonic
seizures with falling). Cognition was assessed after 24 h of the last
PTZ injection. Number of animals in each group was eight.
29
2.4. Cognition and behavioural tests
For the cognitive assessment each animal was assessed at a
time. Cognition was assessed by following two separate beha-
vioural tests.
2.4.1. Elevated plus maze
Elevated plus maze is widely used to assess the acquisition and
retention latency of the animals. The maze was constituted of
wood painted gray and contained a central platform (5 5) cm
with radiated four symmetrical arms: two open arms (16 5) cm
and two closed arms (16 5 10) cmwith an open roof. The maze
is elevated to a height of 25 cm from the oor. On the rst day, the
acquisition transfer latency was noted as follows: the mice were
placed individually at the end of one open arm, facing away from
central platformand the time it took to move fromthe open armto
either of the closed arm was recorded. During training, if the
animal did not enter in closed armwithin 90 s it was gently pushed
in the closed arm. To become acquainted with the maze, the
animals were allowed to explore the maze for 10 s after reaching
the closed arm and then they returned to their home cage. The
animals were retested 24 h after the rst day training and the time
taken to enter the closed arm was taken as retention latency. A
time period of 90 s was taken as cut-off.
34
2.4.2. Passive avoidance response
A derangement in memory was assessed by this test. Briey, an
inverted petridish was placed in the centre of the grid oor of a
N.B. Agarwal et al. / Seizure 20 (2011) 257262 258
continuous avoidance apparatus. The petridish served as the shock
free zone (SFZ). Mice were placed individually on SFZ and while on
stepping down from the SFZ they were given an electric shock of
20 V through the grid oor. Animals were trained to remain on the
SFZ for 60 s. The step-down latency in seconds was recorded after
the last training session without giving shock, this was termed
acquisition latency. The animals were retested after 24 h, without
giving shock and the time taken for the mouse to step down was
measured and termed as retention latency. A cut-off of 600 s was
chosen for the animal which did not step down in this period.
35
2.5. Measurement of oxidative stress parameters
2.5.1. Tissue preparation
At the end of study period, the animals were sacriced by ether
anaesthesia; the brain was quickly dissected out in toto, washed
with ice-cold sodium phosphate buffer, weighed and stored over
ice. The brains were further processed within half an hour of
dissection and the estimation of oxidative stress were done in the
same working day. Brain tissue was homogenized with 10 times
(w/v) sodium phosphate buffer. The homogenate was centrifuged
at 3000 rpm for 15 min. The supernatant was used for the
estimations of MDA, reduced glutathione levels, superoxide
(SOD) and catalase activities.
2.5.2. Estimation of lipid peroxidation
Malondialdehyde (MDA), which is a measure of lipid peroxida-
tion, was measured spectrophotometrically by the method
described by Ohkawa et al.
36
2.5.3. Estimation of reduced glutathione
Reduced glutathione was estimated spectrophotometrically by
the method described by Ellman.
37
2.5.4. Estimation of superoxide dismutase
Superoxide dismutase activity was determined by using the
method proposed by Marklund and Marklund.
38
2.5.5. Estimation of catalase
Catalase activity was determined by using the method
proposed by Claiborne.
39
2.6. Statistical analysis
The results are expressed as mean SEM (standard error of
mean). Statistical analysis was performed using analysis of variance
(ANOVA) with Tukeys post hoc statistical tests. A p < 0.05 was taken
as the level of signicance.
3. Results
3.1. Effect of different drug treatments alone on transfer latency (TL)
in non-kindled mice
None of the administered drugs impaired cognition as
evidenced by no signicant change in acquisition and retention
latencies in the elevated plus maze test (Fig. 1).
3.2. Effect of different drug treatments on transfer latency (TL) in PTZ-
kindled mice
There was no signicant change observed in the TLs in various
groups of animals during acquisition test. However, on the 2nd day
vehicle treated PTZ-kindled group demonstrated signicantly
(p < 0.001) impaired retention TL as compared to control groups
of animals. Further, topiramate (10 mg/kg) treatment was found to
signicantly impair the retention TL when compared to vehi-
cle + PTZ-kindled group. However, lamotrigine and oxcarbazepine
treatment didnot impair the acquisitionlatency as well as retention
latency when compared to vehicle + PTZ-kindled group (Fig. 2).
3.3. Effect of different drug treatments alone on step down latency
(SDL) in non-kindled mice
In all treatment groups no signicant impairment was observed
in acquisition and retention SDLs (Fig. 3).
3.4. Effect of different drug treatments on step down latency (SDL) in
PTZ-kindled mice
PTZ-kindling signicantly impaired the cognition as evidenced
by decreased SDL on the second day of the test. Neither lamotrigine
nor oxcarbazepine impaired the cognition as indicated by no
change in acquisition and retention SDLs in the PTZ-kindled
animals. However, topiramate treatment was observed to produce
a signicant impairment of cognition as evidenced by a
signicantly decreased retention SDL (p < 0.001) (Fig. 4).
3.5. Effect of different drug treatments alone on oxidative stress in
whole brain tissue of non-kindled mice
In chronic study administration of lamotrigine and oxcarbaze-
pine alone did not alter MDA and reduced glutathione levels,
catalase and superoxide dismutase enzyme activities in whole
brain tissues as compared to control group of animals. On the other
hand, topiramate administration was found to increase the MDA
levels as compared to control animals (Table 1).

Fig. 1. Effect of different drug treatment on transfer latency (TL) in non-kindled

mice. Values are expressed as mean SEM, n = 8 in each group, ANOVA followed by
Tukeys test.

Fig. 2. Effect of different drug treatments alone on transfer latency (TL) in kindled
mice.
a
p < 0.001 control vs. PTZ-kindled.
b
p < 0.01 PTZ-kindled + Topiramate vs.
PTZ-kindled.
N.B. Agarwal et al. / Seizure 20 (2011) 257262 259
3.6. Effect of various drugs on oxidative stress levels in kindled mice
brain tissues
In PTZ-kindled mice MDA levels were observed to increase
(p < 0.001) and reduced glutathione levels were found to decrease
(p < 0.001) when compared to the values in control group of mice.
Administration of oxcarbazepine and lamotrigine was also found to
increase the MDA levels and decrease in reduced glutathione levels
but results were not statistically different from those of vehicle
treated PTZ-kindled animals. However, topiramate administration
has further increased MDA levels (p < 0.001) and decreased
glutathione levels (p < 0.01) as compared to the respective values
in vehicle + PTZ kindled mice brain tissue (Table 1).
3.7. Effect on mice brain SOD and catalase activity
No signicant change in SOD and catalase activities was
observed in all the treated groups when compared to control group
(Table 1).
4. Discussion
Clinical reports suggest that not only epilepsy but therapeutic
drugs used in the treatment of epilepsy also exert negative effect
on cognition which affects quality of life of epileptic patients. Many
authors have reported efcacy of certain plants viz. Bacopa
monnieri,
58
Embilica ofcinalis
59
and Centella asiatica
60
and active
constituents viz. curcumin
61,62
against epilepsy and oxidative
stress. Some studies concluded the elevation of oxidative stress
during epilepsy. Treatment with these plants and active consti-
tuents possessing antioxidant property has been shown to improve
cognitive disorders in animals. Recently, an elevation of oxidative
stress has been observed with the treatment of valproate,
carbamazepine and levetiracetam in epileptic patients.
63
Report-
edly, AED offers signicant seizure suppression but impairs
cognition and increases oxidative stress in patients. However, it
is not clear whether it is epilepsy or oxidative stress or both which
contribute to the decline of cognitive function. It has been reported
consistently that older generation antiepileptic drugs cause
cognitive impairment.
40
But, newer agents have been less explored
for their adverse effect prole on cognition during epilepsy
treatment.
1725
The present study was conducted to compare the
effects of three newer AEDs viz. lamotrigine, oxcarbazepine and
topiramate on cognition during experimental epileptogenesis in
mice. Step-down latency and transfer latency on passive avoidance
apparatus and elevated plus tests, respectively were used to assess
cognition. Experimental epilepsy was induced by repeated
administration of pentylenetetrazole in mice.
The results of the present study indicate that (retention)
transfer latency was signicantly increased and (retention) step-
down latency was signicantly decreased in vehicle treated PTZ-
kindled mice as compared to control groups signifying memory
impairment. However, acquisition latencies remained unaffected.
Cognition tests were also performed in the third week but results
showed no impairment (data not shown). Thus it is important to
note that cognition impairment occurred only after the develop-

Fig. 4. Effect of different drug treatments alone on step-down latency (SDL) in

kindled mice.
a
p < 0.001 control vs. PTZ-kindled.
b
p < 0.01 PTZ-kindled+
Topiramate vs. PTZ-kindled

Fig. 3. Effect of different drug treatments alone on step-down latency (SDL) in non-
kindled mice. Values are expressed as mean SEM, n = 8 in each group, ANOVA
followed by Tukeys test.
Table 1
Effect of various drugs on oxidative stress levels in non-kindled and kindled mice brain tissues.
Groups Dose (ml/kg) MDA
(nmole/g wet tissue)
Reduced glutathione
(mg/g wet tissue)
Superoxide dismutase
(U/mg protein)
Catalase
(U/mg protein)
Control 10 191.53.77 1865.18 8.590.39 12.140.29
Lamotrigine 5 23412.35 170.0312.49 8.040.26 12.430.11
Oxcarbazepine 15 207.136.21 164.8811.39 8.180.15 11.510.31
Topiramate 10 262.256.1
a
163.1312.46 8.030.11 11.080.11
Vehicle+PTZ 25 2935.55
b
1333.71
f
8.0250.19 11.290.14
PTZ+lamotrigine 5 282.7523.95
d
103.388.84
h
8.80.15 12.00.17
PTZ+oxcarbazepine 15 252.3836.52
e
108.59.22
i
7.80.26 11.60.2
PTZ+topiramate 10 373.759.53
c
93.52.87
g
8.340.057 11.510.17
Values are expressed as meanSEM, n =8 in each group, ANOVA followed by Tukeys test.
a
p<0.05 control vs. topiramate.
b
p<0.001 control vs. PTZ-kindled.
c
p<0.001 PTZ-kindled vs. PTZ+topiramate.
d
p<0.001 control vs. PTZ+lamotrigine.
e
p<0.001 control vs. PTZ+oxcarbazepine.
f
p<0.001 control vs. PTZ-kindled.
g
p<0.001 PTZ-kindled vs. PTZ+topiramate.
h
p<0.001 control vs. PTZ+lamotrigine.
i
p<0.001 control vs. PTZ+oxcarbazepine.
N.B. Agarwal et al. / Seizure 20 (2011) 257262 260
ment of full kindling. Thus our results are in agreement with
previous experimental data that has clearly depicted that PTZ-
kindling impairs long term memory.
40
Our ndings are also in
agreement with the previous reports where kindling has induced
retrograde amnesia in the shuttle box and step down avoidance
test in rodents.
42
Clinical evidence available so far indicates that
there is impairment of cognition in epileptic patients.
1315,41
Thus
our results are in line with both preclinical data as well as clinical
studies.
All the administered drugs viz., lamotrigine, oxcarbazepine and
topiramate to non-kindled animals did not alter transfer latency as
well as step-down latency in both acquisition as well as retention
sessions thereby suggesting that these agents do not affect normal
cognitive functions.
On contrary, topiramate administration to kindled animals
showed signicant increase in retention transfer and vice versa
decrease in step down latency, suggesting impairment of cognition
as compared to vehicle treated PTZ-kindled animals. These results
indicate that not only kindling but its treatment with topiramate
further culminates into cognitive impairment. Some previously
documented studies support our results where topiramate
administration in epileptic patients has impaired memo-
ry.
18,19,21,22,24
When lamotrigine and oxcarbazepine were admin-
istered to kindled animals, both groups showed no signicant
change in transfer and step-down latencies as compared to vehicle
treated PTZ-kindled mice. These results suggest that cognition
impairment in lamotrigine and oxcarbazepine treated animals
could be due to epileptogenesis itself. While comparing these three
drugs it was clear that lamotrigine and oxcarbazepine treatment
do not cause cognitive impairment. These observations are further
conrmed by the previous studies where lamotrigine and
oxcarbazepine treatment showed no signs of neurobehavioural
adverse effects on cognitive functions in the patients of epilep-
sy.
26,4345,4749,51
Kindling effect is a relatively permanent alteration of brain
functions leading to increased excitability to ineffective stimuli.
Several reports indicate that free radical generation due to the
increased activity of glutamatergic transmitter plays a crucial role
in neuronal cell death of the PTZ-kindled animal. These free
radicals cause cellular dysfunction by attacking the polyunsatu-
rated sites of the biological membranes resulting into increased
lipid peroxidation. The levels of glutathione are also reduced
during oxidative stress.
29,45,46,50,53
Long term use of antiepileptic
drugs have also been shown to increase oxidative stress.
54
On the
background of these data, we have measured four oxidative stress
parameters viz., malondialdehyde, glutathione levels, superoxide
dismutase and catalase activities.
Free radicals are normal products of cellular aerobic metabo-
lism. However, when the production of free radicals increases or
defence mechanism of the body decreases, they cause cellular
dysfunction by attacking at the polyunsaturated sites of the
biological membranes leading to lipid peroxidation. The increase
in levels of MDA is a marker of lipid peroxidation.
29
Lamotrigine
and oxcarbazepine administration to non-kindled animals showed
no change in MDA level indicating that these drugs do not cause
induction of oxidative stress. In contrast topiramate administra-
tion in non-kindled animals signicantly increased MDA level
indicating that topiramate induces oxidative stress.
In kindled animals the results show that MDA levels were
increased in the vehicle treated PTZ-kindled brain tissues of mice.
Lamotrigine and oxcarbazepine administration to kindled mice
showed no signicant change in oxidative stress markers as
compared to vehicle treated PTZ-kindled group. However,
topiramate treatment has further increased the oxidative stress
in the PTZ-kindled mice. These results are in consistent with the
previous studies which showed increased oxidative stress in the
brain of kindled rodents.
29,45
The accentuation of oxidative stress
in topiramate treated kindled and non-kindled animals in our
study is supported by in vitro experiments which demonstrated
that topiramate increases oxidative stress in primary rat astro-
cytes.
51
Further, Cardile et al.
52
have also reported that topiramate
has induced production of reactive oxygen species, increased nitric
oxide synthesis, decreased glutamine synthetase activity and cell
viability in primary cultures of rat cortical astrocytes. Thus,
topiramate could be increasing oxidative stress in both kindled and
non-kindled mice.
Glutathione is an endogenous antioxidant present mainly in the
reduced form within the cells. It reacts with the free radicals and
prevents the generation of hydroxyl radical, which is the most
toxic form of free radicals. During this defensive process, reduced
glutathione is converted to oxidized form with the help of the
enzyme glutathione peroxidase. The biochemical results of the
present study reveal comparable decrease in glutathione levels in
vehicle-, lamotrigine- and oxcarbazepine-treated PTZ-kindled
mice. The results show that this decline is due to kindling itself.
However, topiramate treatment shows signicant decrease in
glutathione levels as compared to vehicle treated PTZ-kindled
group. It is clear that glutathione was depleted while combating
free radicals.
Superoxide dismutase (SOD) is an antioxidative enzyme of the
brain. In the present study there was no change in SOD activities in
all the treatment groups, this could be due to the fact that
glutathione may be playing a crucial role in combating oxidative
stress. Catalase is an enzyme responsible for detoxication of H
2
O
2
formed by the action of superoxide dismutase. The insignicant
change in the activities of catalase might be due to the fact that the
catalase activity in the rodent brain is very low
46,51
and it may not
be the key enzyme involved in intracellular antioxidant defence
system in the brain. The present results further suggest that
topiramate has more oxidative effect via MDA and glutathione
than via superoxide dismutase or catalase activity.
Thus the results of the present study suggest that lamotrigine
and oxcarbazepine do not affect cognition during PTZ-induced
kindling in mice while topiramate deteriorates cognitive functions
by altering biochemical parameters during experimental epilep-
togenesis. It is also evident from the results that oxidative stress
could be causative mechanism involved in cognitive impairment
during experimental epileptogenesis as well as during drug
treatment. Further, clinical and biochemical studies are required
to demonstrate a correlation between cognitive dysfunction and
oxidative stress during epilepsy and antiepileptic drug therapy.
Acknowledgement
The authors are thankful to Sun Pharma, Ahmadabad for
providing drug samples of lamotrigine and oxcarbazepine.
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