Beneficial Interaction of Piperine with Sodium Valproate against maximal Electroshock induced Seizures in Mice

 

Uma Devi P1*, Sooraj Surendran2, Merin Babu2, Jipnomon Joseph2

1Associate Professor, Department of  Pharmacology, Amrita School of Pharmacy, Amrita University, Kochi - 682041, Kerala, India

2Department of Pharmacology, Amrita School of  Pharmacy, Amrita University, Kochi - 682041, Kerala, India

*Corresponding Author E-mail: umadevip@aims.amrita.edu; umadeviaims@gmail.com

 

ABSTRACT:

Modulation of brain oxidant: antioxidant balance has been reported in different animal models of seizures including electroshock induced seizures. Thus, we tried to find out the effect of combining piperine, an antioxidant with sodium valproate against seizures induced by maximal electroshock (MES). Male Swiss albino mice (28-34 g) were assigned to different groups: sodium valproate (150 and 300 mg/kg), piperine (5 and 10 mg/kg) and their combination. Electroconvulsions were produced in mice using a current of 25 mA for 0.2 s. Sodium valproate (150 mg/kg) and piperine (5 and 10 mg/kg), did not significantly reduce the duration of tonic hind limb extension. However, the former at a dose of 300 mg/kg significantly decreased the duration of tonic phase (P<0.05). Further, treatment of sodium valproate (300 mg/kg) in combination with piperine (10 mg/kg) completely abolished the extensor phase in all animals. Combined therapy significantly (P<0.01) attenuated the increase in malondialdehyde levels caused by electroshock reflecting the neuroprotective potential of the combination. To conclude, our preliminary results suggest a beneficial interaction of piperine with sodium valproate against electroshock seizures.

 

KEYWORDS: Sodium valproate; piperine; maximal electroshock; malondialdehyde; tonic phase

 

 


INTRODUCTION:

The last few decades have seen the introduction of many new antiepileptic drugs. However, still nearly 30% of epileptic patients do not respond to the currently available antiepileptic drugs.1-3 Hence, there is an urgent need to develop newer antiepileptic drugs or try novel combination therapies in an attempt to achieve enhanced efficacy. Modulation of brain oxidant: antioxidant balance has been reported in different animal models of seizures including electroshock induced seizures.4,5 Treatment with antioxidants has not only been reported to provide protection against seizures, but has also been reported to enhance the efficacy of antiepileptic drugs when given in combination.6,7

Thus, we tried to find out the effect of combining piperine, an antioxidant and a neuroprotective agent8, with sodium valproate against seizures induced by maximal electroshock (MES). Male Swiss albino mice (28-34 g) were assigned to different groups of 6 mice each. The study utilized the following drugs: sodium valproate (150 and 300 mg/kg) and piperine (5 and 10 mg/kg). Sodium valproate was dissolved in 0.9% normal saline and piperine was suspended in 0.5% carboxymethyl cellulose. All the treatments were given orally for 8 days. On the 8th day, electroconvulsions were produced in mice using a current of 25 mA. This current was delivered for 0.2 s duration from the Electro Convulsive Therapy Unit (Ugo Basile, Italy). The duration of tonic extensor phase was recorded for each mouse. Abolition of tonic extensor phase was the criterion for protection.  Immediately after electroshock, the mice were sacrificed and the brain tissue was removed for estimation of malondialdehyde and protein levels.9,10 The study was undertaken only after approval from the Institutional Animal Ethics Committee (IAEC/2015/3/12). The results are presented as mean ± standard error of mean. The results were statistically analysed with one-way analysis of variance followed by the post hoc Bonferroni’s test for multiple comparisons. The Fisher’s exact probability test was used to analyse the results of percentage protection. Results were considered statistically significant if P<0.05.

Sodium valproate (150 mg/kg) and piperine (5 and 10 mg/kg) administered 90 minutes before the MES test, did not significantly reduce the duration of tonic hind limb extension. However, sodium valproate at a dose of 300 mg/kg significantly decreased the duration of tonic phase (P<0.05). Further, treatment with sodium valproate (300 mg/kg) in combination with piperine (10 mg/kg) completely abolished the extensor phase in all animals (Table 1).


 

Table 1: Effect of sodium valproate, piperine and their combination on electroshock induced seizures

Group

Treatment (n=6)

Dose (mg/kg, orally)

Duration of tonic phase (seconds)

Percent protection

Brain MDA levels (nmoles/mg protein)

A

Vehicle

10 ml/kg

-

-

1.402±0.253

B

Vehicle + MES

10 ml/kg

14.17±1.08

0

5.135±0.606***

C

SVP + MES

150

8.33±2.43

16.7

5.446±0.401

D

SVP + MES

300

5.67±2.59#

50

3.697±0.529

E

PIP + MES

5

11.17±2.47

16.7

4.668±1.11

F

PIP + MES

10

7.33±2.36

33.3

4.914±0.463

G

SVP + PIP + MES

300 + 10

0###

100##

2.072±0.704##

Values are presented as mean ± SEM. n = number of animals; MDA = malondialdehyde; MES = maximal electroshock; PIP = piperine; SVP = sodium valproate. Treatment duration = 8 days. ***P<0.001 versus Group A; #P<0.05, ##P<0.01 and ###P<0.001 versus Group B. Significant by one-way analysis of variance followed by Bonferroni’s multiple comparison test (duration of tonic phase and brain MDA levels) and Fisher’s exact test (percent protection against seizures).

 


In the present study, brain MDA levels were estimated. Following electroshock administration, brain MDA levels were significantly increased (P<0.001). Pre-treatment with sodium valproate and piperine per se did not significantly alter the MDA levels. However, combined therapy of sodium valproate (300 mg/kg) with piperine (10 mg/kg) significantly (P<0.01) attenuated the increase in MDA levels caused by electroshock reflecting the neuroprotective potential of the combination. None of the treatments at the doses tested could alter the brain glutathione levels significantly (data not shown). To conclude, our preliminary results suggest a beneficial interaction of piperine with sodium valproate against electroshock seizures. Additional isobolographic studies are required to characterize the nature of interaction. In view of the modulation of gamma amino butyric acid and sodium channels by piperine11-14, we cannot rule out the involvement of neurotransmitter systems and ion channels in the observed beneficial effects which needs to be probed in further studies.

 

ACKNOWLEDGEMENT:

The financial assistance from the Kerala State Council for Science, Technology and Environment (KSCSTE) to Sooraj Surendran is acknowledged.

 

CONFLICT OF INTEREST:

None.

 

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Received on 24.06.2017             Modified on 18.07.2017

Accepted on 19.09.2017           © RJPT All right reserved

Research J. Pharm. and Tech 2017; 10(11): 3967-3968.

DOI: 10.5958/0974-360X.2017.00720.X