Maiha et al., Nig. Journ. Pharm. Sci., March, 2009, Vol. 8 No. 1, P.

102 – 108

Nigerian Journal of Pharmaceutical Sciences

Vol. 8, No. 1, March, 2009, ISSN: 0189-823X
All Rights Reserved

ANTICONVULSANT STUDIES ON COCHLOSPERMUM

TINCTORIUM AND PAULLINIA PINNATA EXTRACTS IN
LABORATORY ANIMALS
1*
Maiha, B. B., 1Magaji, M. G, 2Yaro, A. H., 1Hamza, A. H, 3Ahmed, S. T and 4Magaji,
R.A.
1
Department of Pharmacology and Therapeutics, Ahmadu Bello University, Zaria
2
Department of Pharmacology, Faculty of Medicine, Bayero University, Kano
3
Federal Government Girls College, New Bussa, Niger state
4
Department of Human Physiology, Faculty of Medicine, Bayero University, Kano

*Author for Correspondence: [email protected]; 234-8062295967

ABSTRACT
Cochlospermum tintorium (Cochlospermaceae) and Paullinia pinnata (Sapindaceae) are widely used
medicinal plants in Northern Nigeria in the management of epilepsy. The hydroalcoholic root bark extract
of Cochlospermum tintorium and methanolic stem bark extract of Paullinia pinnata were investigated for
anticonvulsant activities using Maximal electroshock test in chicks, Pentylenetetrazole and Strychnine-
induced seizures, in mice. The Paullinia pinnata extract produced a dose-dependent protections (50%, 60%
and 70%) against maximal electroshock at doses tested (125, 250 and 500 mg/kg) while the C. tintorium
extract did not protect the animals at the doses tested (5, 10 and 20 mg/kg). The two extracts did not protect
the animals against strychnine-induced seizure. Both extracts protected 20% of the animals against
pentylenetetrazole-induced seizure at the highest doses tested This study suggests that the aqueous
methanolic stem bark extract of P. pinnata contains bioactive constituents that may be beneficial in grand
mal epilepsy and lend pharmacological credence to the ethnomedical claim for the use of the plant in the
management of this epilepsy.

Key Words: Paullinia pinnata; Cochlospermum tintorium; Seizure; Maximal electroshock;

Pentylenetetrazole; Strychnine

INTRODUCTION prevalence of epilepsy in Nigeria is 8 to

Epilepsy is a chronic brain disorder 13 per thousand people (Azubuike and
characterized by recurrent derangement Nkanginieme, 1996). About 30% of
of the nervous system due to sudden epilepsy is refractory to treatment
excessive disorderly discharge from the (WHO, 2001). Many people in
cerebral neurons (Kabir et al., 2005). It developing countries may not receive
is the second most common neurological basic treatment due to high cost,
disorder attended to by neurologists unavailability and untoward effects
(Sridharan, 2002). The estimated associated with the available

102
Maiha et al., Nig. Journ. Pharm. Sci., March, 2009, Vol. 8 No. 1, P. 102 – 108

Antiepileptic drugs. Therefore, many activities of these medicinal plants. This

people in the developing countries still study, therefore, aims at evaluating the
rely on traditional healing practices and anticonvulsant properties of the aqueous
medicinal plants for their basic health methanolic root bark extract of C.
care needs. WHO encourages the tinctorium and stem bark extract of P.
inclusion of herbal medicines of proven pinnata in laboratory animals.
safety and efficacy in the healthcare
programs of developing countries (Amos MATERIALS AND METHODS
et al., 2001). Scientific research is Preparation of plant material and
needed to provide evidences of the extracts
safety and efficacy of beneficial Fresh stem bark of Paullinia pinnata
medicinal plants. Cochlospermum were collected from Sakani village along
tinctorium and Paullinia pinnata are New Jos Road, Kaduna state, Nigeria
commonly used medicinal plants in while fresh root part of Cochlopermum
traditional medicine practice in Nigeria. tintorium was collected from a
Cochlospermum tintorium secondary forest in New Bussa, Niger
(Cochlospermaceae) is a bushy plant state, Nigeria in January 2008. The
attaining about 50 cm in height. It has collections were authenticated by Malam
widespread occurrence in the savannah Musa Mohammed and Umar Galla of the
and shrub land throughout the drier areas Herbarium Section in the Department of
of West Africa region. The common Biological Sciences, Ahmadu Bello
vernacular names in Nigeria include University (ABU), Zaria-Nigeria.
rawaya, kyamba (Hausa), obazi, abanzi Voucher specimens (number 2314 for C.
(Igbo) and sewutu (Yoruba). The root tinctorium and 427 for P. pinnata) were
decoction is used for management of deposited at the herbarium for future
epilepsy (Burkill, 2000). reference. The root bark of C.
Paullinia pinnata (Sapindaceae) is a tinctorium and the stem bark of P.
woody or sub-woody climber at damp pinnata were air dried separately under
site and stream banks of the forest and shade until constant weights were
jungle regrowth in the savannah zone. obtained. They were then size-reduced
The common vernacular names in into coarse powder with a pestle and
Nigeria include goron dorina; yatsa mortar. Extraction was carried out using
biyar (Hausa) edefina; aliligo (Igbo) and cold maceration with occasional shaking
ogbe-okiye (Yoruba). Decoction of the for 72 hr using 500 mL of 70% aqueous
leafy twigs is used in West Africa for methanol for 100 mg of each powdered
jaundice, yellow fever and heart materials. The extracts were
irregularities while the leaves decoction concentrated in vacuo and subsequently
is used for diarrhoea, dysentery and colic referred to as Paullinia pinnata extract
(Burkill, 2000). The root is reported to (PPE) and Cochlopermum tintorium
be used in northern part of Nigeria in the extract (CTE). Fresh aqueous solutions
management of convulsion (Personal of the extracts were prepared for each
Communication: Ado Ibrahim of study.
Science Secondary School, Dawakin
Tofa, Kano State). Experimental animals
To our knowledge, there is no report in
the literature on the anticonvulsant

103
Maiha et al., Nig. Journ. Pharm. Sci., March, 2009, Vol. 8 No. 1, P. 102 – 108

Swiss albino mice (18-30 g) of either sex The first group received normal saline
were obtained from the animal house (10 ml/kg) i.p.; second, third and fourth
facility of the Department of groups were treated with PPE (125
Pharmacology and Clinical Pharmacy, mg/kg, 250 mg/kg and 500 mg/kg), i.p.
Ahmadu Bello University, Zaria, The fifth, sixth and seventh groups
Nigeria. Day old Rangers cockerels were received CTE (5 mg/kg, 10 mg/kg and
obtained from the National Animal 20 mg/kg). The last group was
Production Research Institute (NAPRI), administered with phenytoin (20 mg/kg),
Shika, Kaduna state Nigeria. The mice, i.p. Thirty minutes after pre-treatment,
maintained on standard rodent feed and maximal electroshock was administered
water ad libitum, were housed in to induce seizure in the chicks using Ugo
polypropylene cages at room basile electroconvulsive machine (Model
temperature throughout the study. All 7801) connected to Claude Lyons
experimental protocols were approved stabilizer with corneal electrodes placed
by the University animal ethics on the upper eyelids of the chicks. The
committee. The experiments were shock duration, frequency and pulse
conducted in a quiet laboratory between width were set and maintained at 0.80s,
the hours of 900 h to 1600 h. 200 pulse per second and 0.8ms
respectively. A current of about 90mA,
Acute Toxicity studies: LD50 which produced tonic seizures in 90% of
Intraperitoneal median lethal doses of the control chicks, was used throughout
CTE and PPE were determined using the the study. Seizures were manifested as
method of Lorke (1983). Briefly; the tonic hind-limb extension (THLE). The
method was divided into two phases. In ability to prevent this feature or prolong
the initial phase, 3 groups of three mice the latency and or onset of the THLE
each were treated with the methanol root was considered as an indication of
bark extract of the plant at doses of anticonvulsant activity (Sayyah et al.,
10,100 and 1000mg/kg body weight i.p. 2002).
and were observed for signs of toxicity
and death for 24 hours. In the second Pentylenetetrazole-induced Seizure in
phase, 4 groups each containing one mice
mouse was injected with four more The method of Swinyard et al., (1989)
specific doses of the extract. The LD50 was employed. Forty mice were divided
value was determined by calculating the into eight groups each containing five
geometric mean of the lowest dose that mice. The first group received normal
caused death and the highest dose for saline (10 ml/kg) i.p.; second, third and
which the animal survived (0/1and 1/1). fourth groups were treated with PPE
(125 mg/kg, 250 mg/kg and 500 mg/kg),
Maximum electroshock-induced i.p. The fifth, sixth and seventh groups
convulsion in chick received CTE (5 mg/kg, 10 mg/kg and
The method described by Swinyard and 20 mg/kg). The last group was treated
Kufferberg (1985) as modified by with 20mg phenobarbitone per kg, i.p.
Sayyah et al. (2002) was employed in (positive control). Thirty minutes post
this study. Eighty (80) one day old white treatment, mice in all the groups
cockerels were randomly divided into received 85mg pentylenetetrazole per kg
eight groups each containing ten chicks. s.c. and were observed over a period of

104
Maiha et al., Nig. Journ. Pharm. Sci., March, 2009, Vol. 8 No. 1, P. 102 – 108

30 minutes. Absence of an episode of followed by Dunnet’s test (SPSS-15). A

clonic spasm of at least 5 seconds P value <0.05 was considered
duration indicated a compound’s ability significant.
to abolish the effect of
pentylenetetrazole on seizure threshold. RESULTS
Subcutaneous Strychnine-induced
Seizure in mice The median lethal dose values for PPE
The first group received normal saline and CTE were found to be 2346.42
(10 ml/kg) i.p.; second, third and fourth mg/kg and 118.32 mg/kg, respectively.
groups were treated with PPE (125 PPE produced a dose-dependent
mg/kg, 250 mg/kg and 500 mg/kg), i.p. protections (50%, 60% and 70%) against
The fifth, sixth and seventh groups maximal electroshock test at doses tested
received CTE (5 mg/kg, 10 mg/kg and (125, 250 and 500 mg/kg) while CTE
20 mg/kg). The last group was treated extract did not protect the animals at the
with 20mg phenobarbitone per kg, i.p. doses tested (5, 10 and 20 mg/kg)
(positive control). Thirty minutes post - (Figure 1). There was no significant
treatment, mice in all the groups decrease in the mean recovery time in
received 1.2mg strychnine per kg s.c. the unprotected chicks in both PPE and
The proportion of mice presenting CTE groups (Table 1). The two extracts
convulsions as well as the onset of tonic did not protect the mice against
convulsions was recorded. Abolition of strychnine-induced seizure (Table 2).
tonic extensor jerks of the hind limbs Both extracts protected 20% of the mice
within 30 minutes after strychnine against pentyleneterazole-induced
administration was considered an seizure at their highest doses. While the
indicator that the testing material could standard drug used, phenorbarbitone (20
prevent strychnine-induced convulsion. mg/kg) afforded 100% protection
(Figure 2). There was no significant
Statistical analysis difference in the mean onset of seizure
in mice treated with the extracts when
The results were analyzed for statistical compared with the control (Table 3).
significance using one-way ANOVA

Fig 1: Effect of methanolic extracts of P. pinnata and C. tinctorium on MEST in chick

105
Maiha et al., Nig. Journ. Pharm. Sci., March, 2009, Vol. 8 No. 1, P. 102 – 108

Fig 2: Effect of methanolic extracts of P. pinnata and C. tinctorium on PTZ induced

seizure in mice (n=5) CTE: Cochlospermum tinctorium extract; PPE: Paullinia
pinnata extract; PBT: Phentobarbitone

Table 1: Effect of methanolic extracts of C. tintorium and P. pinnata on MEST in

chicks
Groups Doses Mean Recovery time (mins)
N/Saline 10ml/kg 5.5 ± 0.34
CTE 5 6 ± 0.93
CTE 10 5.2 ± 1.2
CTE 20 4.5 ± 1.27
PPE 125 5.2 ± 0.37
PPE 250 4.5 ± 0.65
PPE 500 11 ± 5.03
CTE: Cochlospermum tinctorium extract; PPE: Paullinia pinnata extract

Table 2: Effect of methanolic extracts of C. tintorium and P. pinnata on Strychnine-

induced Seizure in mice
Groups Doses Quantal Mean onset time
protection (mins)
N/Saline 10ml/kg 0/6 3.0 ± 0.77
CTE 5 0/6 2.8 ± 0.2
CTE 10 0/6 2.0 ± 0.2
CTE 20 0/6 2.0 ± 0.00
PPE 125 0/6 2.2 ± 0.24
PPE 250 0/6 3.0 ± 0.31
PPE 500 0/6 4.0 ± 0.55
PBT 20 6/6 -

CTE: Cochlospermum tinctorium extract; PPE: Paullinia pinnata extract; PBT: Phenorbarbitone

106
Maiha et al., Nig. Journ. Pharm. Sci., March, 2009, Vol. 8 No. 1, P. 102 – 108

Table 3: Effect of methanolic extracts of C. tintorium and P. pinnata on

Pentylenetetrazole-induced seizure in mice
Groups Doses Mean onset time (mins)
N/Saline 10ml/kg 3.0 ± 0.77
CTE 5 6.0 ± 0.00
CTE 10 1.2 ± 0.2
CTE 20 2.0 ± 0.00
PPE 125 1.6 ± 0.2
PPE 250 1.75 ± 0.25
PPE 500 2.25 ± 1.25
CTE: Cochlospermum tinctorium extract; PPE: Paullinia pinnata extract

DISCUSSION effective in the therapy of absence or

The MES and PTZ models are widely myoclonic seizures.
believed to be predictive of activity in Strychnine is a competitive antagonist
common form of human epilepsy of the inhibitory amino acid glycine
(Wickenden, 2002). There is no false (Larson, 1969). The inability of the
negative in MES test and all currently extracts to protect against strychnine-
available AEDs that are clinically induced seizure suggests non-
effective in the management of involvement of glycine receptors in their
generalized tonic-clonic and partial activity.
seizures such as phenytoin, The results of this study suggest that the
carbamazepine and lamotrigine all aqueous methanolic stem bark extract of
suppresses the hind limb tonic extension Paullinia pinnata (PPE) contains
(HLTE) in MES test (Browning, 1992; bioactive constituents which possess
Rho and Sankar, 1999). They act by anti-convulsant acivity that may be
modulating neuronal voltage gated beneficial in the management of grand
sodium channels (Wickenden, 2002). mal epilepsy and lend credence to the
Thus, the ability of a compound to use of the plant in the management of
inhibit HLTE suggests anticonvulsant epilepsy in traditional medicine. Further
activity for the treatment of generalized work is going on in our laboratory to
tonic clonic and partial seizures. determine the constituent(s) responsible
Compound that protect against MES are for these observed effects.
known to prevent the spread of seizure.
Therefore, the ability of the PPE to REFERENCES
inhibit the HLTE suggests Amos, S., Kolawole, E., Akah, P., Wambebe, C.
anticonvulsant activity which may be and Gamaniel, K. (2001). Behavioral effects of
the aqueous extract of Guiera senegalensis in
beneficial in generalized tonic-clonic mice and rats. Phytomedicine, Vol. 8(5): 356-
and partial seizures. 361.
Anticonvulsant activity in PTZ test
identifies compounds that can raise Azubuike, J.C. and Nkanginieme, K.E.O. (1996).
seizure threshold in the brain (White et Paediatrics and child health in a tropical region.
Fourth Dimension Publishers, Enugu, Pp. 388.
al., 1998). The inability of PPE and CTE
to protect the mice against PTZ-induced Browning, R. (1992). The electroshock model,
seizure suggests they may not be neuronal network and antiepileptic drugs. In:
Faingold, C.L. and Fromm, G. H. (Eds) Drugs

107
Maiha et al., Nig. Journ. Pharm. Sci., March, 2009, Vol. 8 No. 1, P. 102 – 108

for control of Epilepsy: Actions on Neuronal Sridharan, R. (2002). Epidemiology of epilepsy.

Networks in seizure Disorders, CRC Press. Boca Current Science, 82 (6); 664-670
Raton, FL. pp. 195-211.
Swinyard, E.A., Kupferberg, H.J. (1985)
Burkill, H.M. (2000). The Useful Plants of West
Tropical Africa. 2nd Edn. Royal Botanical Antiepileptic drugs: detection, quantification and
Garden. Kew, Vol.1: Pp 387; Vol. 5. Pp 26-30. evaluation. Federation Procedings, 44: 3.

Kabir, M., Iliyasu, Z., Abubakar, I.S., Kabir, Z.S. Swinyard, E.A., Woodhead, J.H., White, H.S.
and Farinyaro, A.U. (2005). Knowledge. and Franklin, M.R. (1989). General Principles:
Attitude and Beliefs about epilepsy among adults Experimental selection, quantification, and
in a northern Nigerian urban community. Annals evaluation of anticonvulsants. In:Levy, R.H.,
of African Medicine, 4(3); 107-112. Mattson, B., Melrum, J.K. and Dreifuss, F.E.
(Eds) Antiepileptic Drugs, 3rd edition. Raven
Larson, M.D.(1969). An analysis of the action of Press. New York. pp. 85-103.
strychnine on recurrent ISPS and amino acid
induced inhibitions in the cat spinal cord. Brain White, H.S., Wolf, H.H., Woodhead, J.H. and
Research, 15:185-200. Kupferberg, H.J. (1998). The National Institute
of Health anticonvulsant drug development
Lorke D (1983). A new approach to acute program: Screening for efficacy. In: French, J.,
toxicity testing. Archives of toxicology, 54:275- Leppik, I.E. and Dichter, M.A. (Eds.)
287. Antiepileptic Drug Development: Advances in
Neurology, Vol. 76, Lippincott-Raven
Rho, J.M. and Sankar, R. (1999). The Publishers: Philadelphia; 29-39.
Pharmacologic basis of antiepileptic drug action.
Epilepsia, 40: 1471-1483. WHO (2001). Epilepsy: aetiology, epidemiology
and prognosis. Fact sheet No 165
Sayyah, M., Saroukhani, G., Peirovi, A. and
Kamalinejad, M. (2002). Analgesic and Anti- Wickenden, A.D. (2002).Potassium channels as
inflammatory Activity of the Leaf Essential oil antiepileptic drug targets. Neuropharmacology,
of Lauraus nobilis Linn. Phytotherapy Research, 43:1055-1060.
17:733-736.

108