Behavioural effects of methanolic extract of Chrysanthellum indicum ...

Yaro, et al., Nig. Journ. Pharm. Sci., October, 2007, Vol. 6 No. 2, P. 127 – 133
Nigerian Journal of Pharmaceutical Sciences
Vol. 6, No. 2, October, 2007, ISSN: 0189-823X
All Rights Reserved
BEHAVIOURAL EFFECTS OF METHANOL EXTRACT OF
CHRYSANTHELLUM INDICUM IN MICE AND RATS
1* Yaro, A. H., 2 Anuka, J. A., 3 Salawu, O. A., 2 Magaji, M. G.
1 Department of Pharmacology, Faculty of Medicine, Bayero University, Kano, Nigeria
2 Department of Pharmacology and Clinical Pharmacy, Ahmadu Bello University, Zaria, Nigeria
3 National Institute for Pharmaceutical Research and Development (NIPRD), Abuja, Nigeria
* Author for Correspondence: (yaroabdulng@yahoo.com)
ABSTRACT
The behavioural effects of methanol extract of Chrysanthellum indicum Linn. Vatke were studied on spontaneous
motor activity (SMA), amphetamine and apomorphine-induced stereotype behaviour, pentobarbitone-induced
hypnosis, exploratory activity and haloperidol-induced catalepsy in mice and rats. The Intraperitoneal and per oral
LD 50 values were also estimated in mice. The intraperitoneal and oral acute toxicity values (LD 50 ) in mice were
found to be 288.5 and 2154 mg/kg body weight respectively. The extract significantly decreased spontaneous motor
activity (SMA) and antagonized apomorphine and amphetamine-induced stereotyped behaviour in mice dose and
time dependently. The extract has no effect on the onset of pentibarbitone-induced sleep, but significantly prolonged
the duration of pentobarbitone-induced sleep and enhanced haloperidol-induced catalepsy dose and time
dependently. It also decreased exploratory activity in mice and had no effect on motor coordination. Our results
provided evidence that the methanol extract of Chrysanthellum indicum contains psycho-active substance(s) with
potential antipsychotic properties. Thus, supporting the development of active substances in the methanol extract for
the treatment of psychoses.
Key words: Chrysanthellum indicum, sleep, catalepsy, coordination, exploratory, sedation, psychoses.
INTRODUCTION
Remedies from plants play an important role
in the healthcare of millions of people
(Rukangira, 2001). Population increase,
inadequate drug supply, exorbitant cost of
treatments and side effects of several
conventional drugs have led to increased
emphasis on the use of plant materials as a
source of medicines for a wide variety of
human ailments including behavioural
disorders.
Majority of the plants used in traditional
medicine lack scientific verification. Many
medicinal plants are used in various ways
because of their activities on the CNS.
However, only limited efforts have been
made to evaluate the potentials of such
plants for their use in modern medicine or to
scientifically justify their traditional use in
the treatment of CNS disorders.
Chrysanthellum indicum Linn. Vatke
(Compositae) is a faintly aromatic herb that
is widely distributed in the tropics. The plant
is commonly known in Hausa as rariyar
kasa (Kontagora), dunkufe (Zaria) and
Goshin ba’ana and in southern Nigeria as
oyigi or abilere in Yoruba (Dalziel, 1955).
Locally in Likoro village of Kaduna state of
Nigeria, the plant is used for the treatment of
mental illness and convulsion (Adamu
Mohammed, personal communication).
The plant has been screened for a
number of pharmacological activities
including anti-tumour activity (Woo et al.,
1977), antiamoebic, diuretic and
127

Yaro, et al., Nig. Journ. Pharm. Sci., October, 2007, Vol. 6 No. 2, P. 127 – 133
hypoglycemic activities (Dhar et al., 1973),
antioxidant (Brasseur et al., 1987) and
gastrointestinal (Amos et al., 2001)
activities. To our knowledge, there is no
report on the behavioural activities of the
plant. This study was designed to test the
behavioural properties of the methanol
extract.
MATERIALS AND METHODS
Collection of plant materials
The whole plant was collected from Likoro
Village, in Kudan Local Government Area,
Kaduna State, Nigeria, in September, 2006.
The plant was identified and authenticated
by staff of the Herbarium Section of
Department of Biological Sciences, Ahmadu
Bello University, Zaria. A voucher
specimen (No. 3110) was deposited at the
herbarium for future reference.
Preparation of extract
The plant material was cleaned, air dried for
7 days and then crushed into coarse powder
with a pestle and mortar. About 100 g of the
powered plant material was successively
macerated with methanol for 48 hours with
occasional shaking. The macerate was
concentrated in vacuo to afford an average
yield of 17.7%.w/w. It was subsequently
referred to as the extract (CI).
Animals
Swiss albino mice (18-25 g) and Adult
Wistar rats (180-220 g) of either sex
obtained from the Animal House Unit,
Department of Pharmacology and Clinical
Pharmacy, Ahmadu Bello University, Zaria
were used. The animals were maintained in
a well ventilated room, fed on Excel feeds
(Feed Masters, Ilorin) and water ad libitum.
All experimental protocols were approved
by the University animal ethics committee.
Phytochemical Test
The extract was screened for the presence of
alkaloids, glycosides, tannins, saponins and
flavonoids according to standard procedure
(Trease and Evans, 1989).
Acute Toxicity Studies in Mice
LD 50 determination was conducted using the
method previously described by Lorke
(1983) for oral and intraperitoneal routes in
mice.
Spontaneous Motor Activity Testing in
Mice
The mice (n=6) were treated with either the
extract (12.5, 25 and 50 mg/kg) or normal
saline. Thirty minutes later, the mice were
transferred individually to Letica Activity
Cages (LE886) connected to a multicount
(LE3806), and after 1min. latency period,
activity counts were recorded for 6 minutes
(Wambebe et al., 1997) at 30, 60, 90, and
120 min.
Amphetamine-induced Stereotyped
Behaviour in Mice
The mice (n=6) were treated with the
extract (12.5, 25 and 50 mg/kg), normal
saline (10 ml/kg) or chlorpromazine (2
mg/kg). Thirty minutes later each mouse
received 2mg amphetamine per kg
intraperitoneally. The signs of stereotyped
behaviour (jumping/climbing, limb licking
and sniffing) were recorded for a period of 2
hours (Ellinwood et al., 1973).
Apomorphine-induced Stereotyped
Behaviour in Mice
The mice were (n=6) pretreated with either
the extract (12.5, 25 and 50 mg/kg) or
normal saline. Thirty minutes later, 2mg
apomorphine per kg, i.p. was administered
to the mice in all the groups. The mice were
observed for signs of stereotypic behaviour
(sniffing, jumping/climbing, and paw
lickings) for two hours using hand tallies
(Nemeroff, 1980).
Pentobarbitone Sleeping Time in Mice
The test was carried out in four groups of six
mice each. The first three groups received
12.5, 25.0 and 50.0mg extract per kg i.p.
respectively, while the last group received
normal saline (10 ml/kg) as control. Thirty
minutes later, 30mg pentobarbitone sodium
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Yaro, et al., Nig. Journ. Pharm. Sci., October, 2007, Vol. 6 No. 2, P. 127 – 133
per kg i.p. was administered to each mouse
to induce sleep. Each mouse was observed
for the onset and duration of sleep, with the
criterion for sleep being loss of righting
reflex (Rolland et al., 1991) while the
interval between the loss and the recovery of
righting reflex was regarded as the duration
of sleep (Fujimori, 1965)
Haloperidol-induced Catalepsy in Rats
Adult Wistar rats were grouped into four
(n=6) and pretreated with either the extract
(12.5, 25 and 50 mg/kg) or normal saline (10
ml/kg). Haloperidol at a dose of 2mg per kg
i.p. was administered to the rats in each
group thirty minutes after pretreatment with
the extract and normal saline. The severity
of catalepsy in each rat was measured every
30 min for 180 min (3 hours). Catalepsy of
an individual rat was measured in a stepwise
manner by a scoring method as follow:
Step I: Each rat was taken out of the cage
and placed on a table. It was then pushed
forward by a gentle touch on the back. If it
failed to move when touched gently on the
back or pushed, a score of 0.5 was assigned.
Step II: the front paws of the rat were
placed alternately on a 3cm high block. If
the rats failed to correct the posture within
15 seconds a score of 0.5 for each paw was
added to the score of step I.
Step III: the front paws of the rats were
placed alternately on a 9cm high block. If
the rat failed to correct the posture within 15
seconds, a score of 1 for each paw was
added to the scores in I and II Thus for an
animal, the highest score was 3.5 (cut-off
Score) and this reflects total catalepsy
(Khisti et al., 1997).
Test for Exploratory Behaviour in Mice
The method used was as described by File
(1973) and modified by Yemitan et al.
(2001). The mice were grouped into 5
(n=6). The apparatus used was a white
painted wooden board (40cm x 40cm) with
four equidistant holes (1cm diameter x 2cm
depth). The animals were treated with the
vehicle (normal saline), the extract (12.5, 25
and 50 mg/kg) or diazepam (1 mg/kg), i.p.
Thirty minutes later, each mouse was placed
at one corner of the board and the number of
head dips recorded over a period of 7.5 min
(File and Wardill, 1975).
Rota-rod Test for Motor Coordination
A rota-rod treadmill device (Ugo Basile N0.
7600, Varese, Italy) was used for this study.
Mice were trained to remain on slowlymoving
(16 revolutions/min) rods of 5cm
diameter for 150 seconds. They were
subsequently grouped into four (n=6) and
treated with either the extract (12.5, 25 and
50 mg/kg) or normal saline (1ml/kg). 30
minutes post-treatment; animals were placed
on the rod at intervals of 30 minutes, up to 2
hours. If an animal failed more than once to
remain on the rod for 3 minutes, it is
considered to lack motor coordination.
Statistical Analysis
Results were expressed as mean ± standard
error of mean; Student’s t-test was used to
determine level of significance of all results
obtained. Results were regarded as
significant at P< 0.05.
RESULTS
The phytochemical screening of the extract
revealed the presence of flavonoids, tannins,
glycosides, alkaloids and steroids.
The extract (12.5 – 50 mg/kg i.p.)
caused a significant (P

Yaro, et al., Nig. Journ. Pharm. Sci., October, 2007, Vol. 6 No. 2, P. 127 – 133
sleep, but significantly (P

Yaro, et al., Nig. Journ. Pharm. Sci., October, 2007, Vol. 6 No. 2, P. 127 – 133
Table 4: Effect of Methanol Extract of C. indicum on Pentobarbitone-induced
sleeping Time in Mice
Treatment (mg/kg) Onset of sleep(min) Duration of Sleep (min)
Normal Saline 4.3 ± 0.2 51.3 ± 6.8
C.I. (12.5) 3.6 ± 0.5 57.8 ± 3.1
C.I(25.0) 4.0 ± 0.3 71.8 ± 5.0 a
C.I.(50.0) 3.6 ± 0.2 101.2 ± 5.5 c
Diazepam (1) 3.2 ± 0.3 b 117.3 ± 4.6 c
All the groups received Pentobarbitone sodium 30mg/kg i.p., Data Presented as
Mean ±SEM, student’s t-test. n = 6, a, and c Are significantly different from
control at P

Yaro, et al., Nig. Journ. Pharm. Sci., October, 2007, Vol. 6 No. 2, P. 127 – 133
play an important role in sleep mechanism.
It is probable that the extract prolonged the
duration of pentobarbitone-induced sleep in
mice via dopaminergic pathways or some
other mechanisms that are related to sleep.
The extract exacerbated haloperidolinduced
catalepsy in rats. Catalepsy is a
trance-like state of self hypnotic sleep
during which there is long-term maintenance
of an animal in an abnormal posture.
Although, neuroleptic-induced catalepsy is
primarily due to the blockade of
dopaminergic
neurotransmission
(Baldessarini, 1990), a number of other
neurotransmitter systems indirectly
influence this response. Potentiation of
haloperidol-induced catalepsy by the extract
in this experiment may well be a simple
synergistic effect of the extract and
haloperidol. The hole-board experiment is a
measure of exploratory behaviour in animals
(File and Wardill (1975). A decrease in this
parameter reveals a sedative behaviour (File
and Pellow, 1985), which has also been
accepted as a parameter for the evaluation of
anxiety conditions, in animals (Crawley,
1985). Reduction of exploratory behavior
without undue sedation, induction of
cataleptic state, inhibition of intracranial
self-stimulation of reward areas, and
prevention of apomorphine-induced
vomiting are other tests that may predict
antipsychotic action (Potter and Hollister,
2004). The extract had no effects on motor
coordination in the treadmill experiment,
suggesting that inhibitory effects observed in
the other studies might be elicited centrally
and not due to a peripheral neuromuscular
blockade. The therapeutic benefits of
traditional remedies might depend upon a
combination of constituents. Some of the
constituents found in this extract might have
contributed to the observed effects. For
instance, alkaloids and saponins have been
reported to show potent sedative activity,
they have also been shown to have
antagonistic activity on amphetamine and
known to inhibit spontaneous motor activity
in mice (Dubois et al., 1986; Taesotikul et
al., 1998). It is therefore, likely that the
alkaloids and saponins content of this plant
might be contributing in part to these
pharmacological effects of the extract.
It may therefore be concluded, based
on the data presented, that the use of
Chrysanthellum indicum in traditional
medicine in Nigeria and other West African
countries is justifiable scientifically. The
pharmacological activities of its active
principles (flavonoids, alkaloids, tannins,
and saponins) correlate very well with those
of antipsychotic agents (e.g. haloperidol).
Further research will involve the isolation of
bioactive components responsible for the
observed pharmacological activities.
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