Facilitatory effect of Piperine on the Anticonvulsant effect of Sodium valproate against Pentylenetetrazole induced Seizures in mice


Sooraj Surendran, Merin Babu, Jipnomon Joseph, Uma Devi Padma*

Department of Pharmacology, Amrita School of Pharmacy, Amrita Vishwa Vidyapeetham,

Kochi - 682041, Kerala, India

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



The present study was designed to find out the effect of piperine as an adjunctive therapy with sodium valproate against seizures induced by pentylenetetrazole (PTZ). Swiss albino mice (25-35g) received either vehicle, sodium valproate (300mg/kg), piperine (10mg/kg) or their combination orally for 8 days prior to PTZ (60mg/kg i.p.) administration. Sodium valproate significantly (P<0.05) increased the latency for clonic generalized seizures. Treatment with sodium valproate and piperine delayed the onset to both jerks (P<0.05) and clonic generalized seizures (P<0.001). All the animals in the combination group were protected from clonic generalized seizures. Our results suggest a favorable interaction of piperine with sodium valproate against PTZ induced seizures.


KEYWORDS: Clonic generalized seizures, pentylenetetrazole, piperine, sodium valproate.




A growing body of evidence from animal studies highlights the modulation of oxidant: antioxidant balance in brain during seizures.1 Inspite of the availability of a number of antiepileptic drugs, very few patients with refractory epilepsy can be made seizure free.2,3 Therefore, there is an urgent need to develop newer novel antiepileptic drugs or explore different combinations of the existing antiepileptic drugs, in an attempt to achieve better seizure control with minimal adverse effects. Addition of antioxidants to sub therapeutic doses of antiepileptic drugs has been reported to produce beneficial effect in experimental seizure models.4 Thus, we tried to find out the effect of piperine, an antioxidant and a neuroprotective agent5, as an adjunctive therapy with sodium valproate, a widely used broad spectrum antiepileptic drug against seizures induced by pentylenetetrazole (PTZ).





Swiss albino mice (25-35g) received either vehicle, sodium valproate (300mg/kg), piperine (10mg/kg) or their combination. All treatments were given orally for 8 days. On the 8th day, immediately after PTZ (60mg/kg i.p.) administration, the time to onset of first jerk and clonic generalized seizure was recorded. The latency was taken as 1800 seconds in the absence of seizures within 30 minutes.6,7 The dose of PTZ was selected as 60mg/kg i.p. based on pilot experiments. This was the dose that produced jerks, myoclonus and clonic generalized seizures in 100% of control animals without mortality. The study was undertaken only after approval from the Institutional Animal Ethics Committee (IAEC/2015/1/19). In view of the role of oxidative stress in the pathology of epilepsy, brain malondialdehyde (MDA) levels were estimated as previously described.8 Brain glutathione levels were also determined as per previously described procedures.9,10  The latency to jerks and clonic generalized seizures is presented as median (range). P<0.05 was considered to be statistically significant.



The latency for the first jerk was not altered significantly by either piperine or sodium valproate. However, the latter significantly (P<0.05) increased the latency for clonic generalized seizures. Further, treatment with sodium valproate in combination with piperine delayed the onset to both jerks (P<0.05) and clonic generalized seizures (P<0.001). All the animals in the combination group were protected from clonic seizures (Table 1).


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


Treatment (n=6)

Dose (mg/kg, orally)

Latency to jerk (seconds)

Latency to clonic generalized seizure (seconds)

Percent protection against clonic generalized seizures



10 ml/kg

107.5 (87-140)

168.5 (127-670)





110.5 (96-192)

1244 (392-1800)*





108.5 (85-152)

403.5 (114-1800)




300 + 10

990 (117-1800)*£#

1800 (1800)***#


Values are presented as median (range). n = number of animals; PIP = piperine; PTZ = pentylenetetrazole; SVP = sodium valproate. Dose of PTZ = 60 mg/kg i.p. Treatment duration = 8 days. * P < 0.05, ** P < 0.01 and *** P < 0.001 versus Group A; £ P<0.05 versus Group B; # P<0.05 versus Group C. Significant by one-way analysis of variance followed by Tukey’s multiple comparison test (latencies) and Fisher’s exact test (percent protection against clonic generalized seizures).


Brain MDA levels were significantly increased but no significant change was observed in the brain glutathione levels following PTZ administration (data not shown). However, none of the treatments at the doses tested could alter the brain MDA or glutathione levels significantly indicating that probably the observed beneficial pharmacodynamic interaction might not be mediated by the antioxidant mechanisms. Considering the fact that certain brain regions may be more susceptible to oxidative insult, we feel that it is essential to determine the oxidative stress markers including the antioxidant defences in different brain regions before completely ruling out their role in the observed effect. Secondly, piperine’s anti-seizure effects have been linked to its modulation of gamma amino butyric acid and serotonin levels in specific brain regions.11-13 Recently, piperine has also been reported to have inhibitory effects on sodium channels.14 Piperine is a promising agent that has been reported to enhance the pharmacological effect of different neurological agents.15-17 Thus, it appears imperative to study the effect of this combination on different antioxidant defences, neurotransmitter systems and ion channels to get a better insight into the mechanisms for the observed beneficial effect.



To conclude, our results suggest a facilitatory effect of piperine on the anticonvulsant effects of sodium valproate against PTZ seizures. However, the molecular basis for the observed effect needs to be explored in further studies.



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



All authors have none to declare.



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Received on 22.06.2019           Modified on 18.07.2019

Accepted on 30.08.2019          © RJPT All right reserved

Research J. Pharm. and Tech 2020; 13(2):651-652.

DOI: 10.5958/0974-360X.2020.00124.9