International Journal of Pharma and Bio Sciences V1(2)2010

ANTICONVULSANT ACTIVITY OF FLOWER PART OF N. ODERUM

POOJA SAINI*1, N. KANNAPAN3, ANUPAMA DIWAN4, PARVEEN KUMAR2, VISHAL ANTIL2,
SHREYA SHARMA2 AND SANDEEP SINGH4
1
R.K.S.D.College of Pharmacy, Kaithal, India
2
G.V.M. College of Pharmacy, Sonipat, India
3
Annamalai University, Chidambaram, TamilNadu, India
4
Hindu College of Pharmacy, Sonipat, India

*Corresponding author [email protected]

ABSTRACT
Different extracts (Petroleum ether, methanolic, aqueous) of Nerium oderum at a dose level of 400 mg/kg
were screened for the anticonvulsant activity using Maximum Electroshock Induced Seizures & Pentylene Tetrazole
Induced Seizures test models. From the present study, Petroleum ether extract of N. oderum showed better
anticonvulsant activity as compared to other extracts.

KEY WORDS
Nerium oderum, Anticonvulsant, Pentylene Tetrazole Induced Seizures and Maximum Electroshock Induced
Seizures

INTRODUCTION and seeds contain cardiac glycosides that have a

Nerium oderum commonly known as “kaner” paralyzing action on the spinal cord. Oleandrin, a pure
belonging to family “Apocynaceae” is provoked as a component from the plant has a stimulating action on
toxic plant but traditionally is being used to cure various the heart and also a pronounced diuretic effect. The
ailments such as asthma, corns, cancer and epilepsy. A alcoholic extract shows antibacterial activity and oil
wide spectrum of biological activities have been obtained from the root is used in leprosy and skin
reported with various constituents isolated from diseases [1]. The plant is recognized in folk medicine as
different parts of the plant. Two varieties are found in antidote, antibacterial, antileprotic, anticancer,
the plant one with the white flowers (Nerium indicum) cardiotonic and C.N.S. depressant [2-4]. In ethno
and one with the pink flowers (Nerium oderum). This is botanical literature, it is mentioned to be effective in the
an erect, smooth shrub 1.5 – 3 meters in height and treatment of cancer, corns & epilepsy and also used as
contains a cream colored, stick, resinous juice. The C.N.S. depressant but no scientific data is reported.
leaves are mostly in whorls of 3 to 4, linear lanceolate. Therefore, in the present study the flowers of the plant
The fruit is cylindrical, in pairs with deep linear are screened for anticonvulsant activity.
striations, slightly twisted & 15-20 centimeters long. MATERIALS AND METHODS
The flowers are showy sweet scented, single or double Collection of Plant Material & Preparation of the
4-5 centimeters in diameter, white, pink or red and Plant Extract
borne on terminal inflorescences (Cymes). Root, bark The flowers of Nerium oderum were collected
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 International Journal of Pharma and Bio Sciences V1(2)2010

ANTICONVULSANT ACTIVITY OF FLOWER PART OF N. ODERUM

and authenticated by Dr. Gyanendra Tiwari. A voucher Phenytoin 20 mg/Kg i.p. and remaining groups received
specimen was deposited in the Department of the test drug at a dose of 400 mg/Kg. After 60 min.,
Pharmacognosy, B.R. Nahata College of Pharmacy, electroshocks (150 mA for 0.2 sec.) were applied by
Mandsaur, India. The collected flowers were dried; means of stainless steel pinna electrodes. The extensor
powdered and successive solvent extraction was done phase of convulsion process was observed.
with Petroleum ether (60-65° C), methanol and water.
The solvents (Petroleum ether, methanol and water) Assessment of anticonvulsant activity by Pentylene
were recovered by the distillation process. Percentage tetrazole (PTZ) induced seizures [6, 7, 8]
yields (% w/w) of different extracts of N. oderum Mice of either sex with a body weight between
obtained by successive solvent extraction were 18 and 22 g have been used for the study. Control group
determined and found to be 4.424, 55.487, 25.44 for received 1 % Tween solution whereas the standard
petroleum ether, methanol and aqueous respectively. group received Diazepam 4 mg/Kg i.p., remaining
These three extracts were then tested for the groups received the test drug at a dose level of 400
anticonvulsant activity. mg/Kg. The test compound was given orally to groups
of 5 mice. After 30 min 60 mg/Kg PTZ (Metrazol) was
Experimental Animals injected i.p. Each animal was placed into an individual
Wistar rats and Male Swiss albino mice were plastic cage for observation lasting 1 h. Seizures and
obtained from Animal House unit of the Department of tonic-clonic convulsions have been recorded.
Pharmacology, B.R. Nahata College of Pharmacy,
Mandsaur (M.P.). The animals are housed in groups of Statistical analysis
5-6 under standard laboratory conditions (temp. Data obtained from pharmacological
25±2°C, relative humidity 55±5% and lighting 08:00- experiments are expressed as mean ± S.E.M (Standard
20-00 hr.) with food & water ad libitium. Error mean). Difference between the control and the
treatments in these experiments was tested for
Acute toxicity study significance using ANOVA followed by Dunnett’s test.
Acute oral toxicity study was performed as per All statistical analysis was performed with prism 4.0
the Organization of Economical Cooperation and (Graph pad software Inc., San Diego, CA). P< 0.05 was
Development (OECD) 425 guidelines [5] .Wistar strain considered significant.
albino rats of either sex were used for the study. The
animals were kept on fasting overnight and were RESULTS
provided only water, after which the extracts (Petroleum Acute toxicity studies depicted no mortality upto
ether, Methanol, aqueous) were administered orally at the dose level of 2000 mg/Kg body weight. So, the
the dose level of 2000mg/ Kg and observed for 14 days. extracts can be considered safe for long term
If no mortality was observed, then the extracts were administration. In PTZ induced seizure test, the onset
found to be safe. of convulsions in control group was observed at 130.6
sec. after PTZ injection. The test drug delayed the onset
Assessment of anticonvulsant activity by Maximum of convulsions to 170.8 sec. (N. oderum, pet. ether
electroshock (MES) induced seizures [6, 7, 8] extract) (Table-1). In supra Maximal Electroshock
Animals are divided into 5 groups, each group Seizure (MES) test, it was observed that the test drug
consisting of 5 animals. Control group received 1 % (N. oderum, Petroleum ether extract) produced a
Tween solution whereas standard group received
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 International Journal of Pharma and Bio Sciences V1(2)2010

ANTICONVULSANT ACTIVITY OF FLOWER PART OF N. ODERUM

significant reduction in the duration of extensor phase control group this duration was 16.20 sec. (Table 2)
which was reduced to 6.6 sec. (P< 0.05), while in

Table 1.
Anticonvulsant activity of different extracts of N. oderum by P.T.Z. induced seizures

S.No. Treatment Onset of convulsion (sec.) Protected animals (%)

1 Control 130.6 ± 4.02 20

2 Standard 187.0 ± 2.56** 100
3 Pet. Ether 170.8 ± 2.74** 100
4 Methanol 156.4 ± 4.02** 80
5 Aqueous 166.4 ± 3.31** 80

Values are expressed in mean ± S.E.M., n = 5, ** Significant at p < 0.01 Vs control, * Significant at p <0.05 Vs control Dunnet’s
test, dose of extracts = 400 mg/Kg, Standard (Diazepam) = 4 mg/Kg

Table 2.
Anticonvulsant activity of different extracts of N. oderum by M.E.S. induced seizures

S.No. Treatment Duration of tonic hind Incidence of convulsions

limb extension(sec.) in no. of mice
1 Control 16.20±0.86 2/5
2 Standard 9.60±0.50** 5/5
3 Pet. Ether 6.6±0.50** 4/5
4 Methanol 12.40±0.92* 2/5
5 Aqueous 11.60±0.50* 3/5

Values are expressed in mean ± S.E.M., n = 5, ** Significant at p < 0.01 Vs control, * Significant at p <0.05 Vs control Dunnet’s
test, dose of extracts = 400 mg/Kg, Standard (Phenytoin) = 20 mg/Kg

DISCUSSION AND CONCLUSION activity against generalized tonic-clonic seizures. N.

Acute toxicity studies indicate that the flower oderum (Petroleum ether extract) was active against
extract of N. oderum may be safely used in the MES induced seizures. PTZ is the most frequently
animals upto the dose level of 2000 mg/ Kg body used substance as well as an acute experimental
weight. The Petroleum ether extract of N. oderum model in the preliminary screening to test potential
significantly inhibited the PTZ and MES induced anticonvulsant drugs. Several biochemical hypothesis
convulsions. It was effective against MES induced have been advanced involving the inhibitory
seizures, since inhibition of the MES test predicts GABAergic system and the system of the excitatory
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 International Journal of Pharma and Bio Sciences V1(2)2010

ANTICONVULSANT ACTIVITY OF FLOWER PART OF N. ODERUM

amino acid glutamate and aspartate [9].The 14 (2004).
mechanism by which PTZ is believed to exert its [6] S. K. Kulkarni. Handbook of Experimental
action is by acting as an antagonist at the GABAA Pharmacology. pp. 133 (2005).
receptor complex [10]. Drugs protecting against [7] G. S. Achilya, S. G. Wadodkar, A. K. Dorle
tonic-clonic seizures induced by PTZ are considered et al. Indian journal of Pharmacology. pp. 33-
to be useful to control myoclonic and absence 36 (2005).
seizures in humans [11]. In this study’s experiments, [8] H. G. Vogel. Drug discovery and evaluation.
the extract of N. oderum replicated the effect of this 2nd ed., pp. 422, 487 (2002).
anti epileptic drug by delaying tonic convulsion and [9] R. I. McDonald, K. M. Kelly. Antiepileptic
mortality. The benzodiazepine site in the GABAA drugs: Mechanisms of action. Epilepsia 34:
receptor and T-type Ca2+ currents could be targets for S1-8-20 (1993).
future studies to know the mechanisms of action of [10] R. Ramanjaneyulu, M. K. Ticku. Interactions
N. oderum (Petroleum ether) extract. It suggests that of pentamethylenetetrazole and tetrazole
N. oderum is useful in suppressing generalized tonic- analogues with thepicrotoxinin site of the
clonic seizures. Anticonvulsant activity of N. oderum benzodiazepine-GABA receptor ionophore
in inhibiting seizures may be by regulating GABA complex. Eur.J.Pharmacol. 98: 337-345
mediated synaptic inhibition through action at (1984).
distinct sites of the synapse. Summarizing the data [11] W. Loscher, D. Schmidt. Which animal’s
obtained in this study, the results suggest a possible models should be used in the search for new
anticonvulsant effect of N. oderum. antileptic drugs? A proposal based on
experimental and clinical consideration.
Epilepsy Res 2: 145-181 (1988).
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