CENTRAL NERVOUS SYSTEM ACTIVITY OF

1
CENTRAL NERVOUS SYSTEM ACTIVITY OF LOTUS
CORNICULATUS VAR. ALBINUS Can Ö D1, Demir Ü 1,
Mor F 2, Öztürk Y11Anadolu University, Faculty
of Pharmacy, Department of Pharmacology, 26470
Eskisehir, TURKEY 2 Dept. Pharmacology and
Clinical Toxicology, Akdeniz University, Burdur,
TURKEY
I. INTRODUCTION The genus Lotus contains
approximately 100 species, distributed in the
Mediterranean area, Eurasia, America, Australia
and China. Lotus corniculatus var. albinus
(Leguminosae) is a common flowering plant native
to grassland temperate Eurasia and North Africa.
The plant is generally known as Birdsfoot Trefoil
because of its birds foot like fruits appear
from July to October (Zhi and Yizhi, 1998). All
parts of the plant are poisonous, containing
cyanogenic glycosides (hydrogen cyanide) (Frohne
and Pfander, 1983 Cooper and Johnson, 1976). In
small quantities, hydrogen cyanide has been shown
to stimulate respiration and improve digestion,
it is also claimed to be of benefit in the
treatment of cancer. In excess, however, it can
cause respiratory failure and even death. This
species is polymorphic for cyanogenic glycosides.
The flowers of some forms of the plant contain
traces of prussic acid and so the plants can
become mildly toxic when flowering. They are
completely innocuous when dried (Duke and Ayensu,
1985). As well as the economic importance of the
plant as forage and green manure, it also has
etnopharmacological importance (Gebrehiwot and
Beuselinck, 2001). Traditionally, Lotus
corniculatus has been used in folk medicine for
its febrifuge, cardiotonic, hypoglycemic,
sedative, vermifuge, carminative, antispasmodic,
restorative and tonic effects (Duke and Ayensu,
1985 Stuart, 1976). The plant is used externally
as a local anti-inflammatory compress in all
cases of skin inflammation (Chiej,
1984). Traditionally, Lotus corniculatus has been
used in folk medicine for its sedative effects in
several countries including Turkey (Baytop, 1999
Chiej, 1984). To the best of our knowledge, there
is no report about the central nervous system
(CNS) activity of this plant. So, the aim of our
study was to investigate the possible CNS
activity of Lotus corniculatus plant extract.
Number of horizontal and vertical movements were
reduced after the application of the extract in
activity cage measurements (Figure 2, Figure 3).
On the other hand, the extract failed to inhibit
motor coordination in Rota-Rod tests (Figure 4).
Figure 2 Effect of 100 mg/kg plant extract on
the total number of horizontal movements, plt0,05,
n7
Figure 3 Effect of 100 mg/kg plant extract on
the total number of vertical movements, plt0,05,
n7
II. MATERIALS AND METHODS II.I.Preparation of
the Extraction Extraction procedures were
applied as it was proposed elsewhere (Tsao and
Den, 2004). Dried aerial parts of the Lotus
corniculatus were ground and extracted with
methanol in Soxhlet apparatus for eight hours.
The extract was concentrated to dryness in rotary
evoporator. Total extractable material with
methanol was found to be 37.97 . II.II.Animals
Swiss albino mice of both sexes, weighing 2530
g, were housed at a room temperature of 18-250C
with a 12-h lightdark cycle (light on 800 AM to
800 PM). Food and water were available ad
libitum. II.III.Behavioral Tests Hole-Board
Test Exploratory behavior of mice was examined 30
minutes after the intraperitoneal (i.p)
application of control (saline) and the 100 mg/kg
plant extract. The board (Ugo Basile, Varese,
Italy) was made of perspex panels and had sixteen
holes with a diameter of 3 cm, which were equally
spaced in the floor. Head-dipping was measured
for 5 minutes under white light (Fiore et al.,
1998). The behavior observed was the total number
of head-dipping during the test. Tail suspension
test Depression levels of mice were examined 30
minutes after the i.p application of saline and
the 100 mg/kg plant extract. The procedure used
was similar to that described by Steru et al.
(1985). Mice were suspended by their tail using
adhesive tape placed approximately 1 cm from the
tip of the tail attached to a wood applicator
stick and hung approximately 30 cm above a table.
The duration of immobility was scored manually
during the last 4 min of a 6-min test. Mice were
considered immobile only when they hung
passively. Activity Cage Test Spontaneous
locomotor activity was assessed in an IR
beam-array activity cage (Ugo Basile, Varese,
Italy). 30 minutes after the i.p injection of
control and the 100 mg/kg plant extract., mice
were individually placed in the center of the
cage and left to freely move for 5 min. The
number of vertical and horizontal crosses of the
IR net is registered for 5 minutes (Pirondi et
al, 2005). Rota-Rod Tests Motor coordination of
mice was examined 30 minutes after the i.p
application of saline and the 100 mg/kg plant
extract. The Rota-Rod (Ugo Basile, Italy) tests
balance and coordination and comprises a rotating
drum, which is accelerated from 4 to 40 rpm. The
time, at which each animal fell from the drum,
was recorded automatically. Each animal received
three consecutive trials and the longest time on
the drum was used for analysis. II.IV.Statistics
The results are presented as mean standard
error of mean and statistical significance was
determined by paired-sample t-test. Statistical
significance was considered when p lt 0.05.
Figure 4 Effect of 100 mg/kg plant extract on
motor coordination of mice, plt0,05, n7
Administration of the extract induced a
significant decrease in the immobility time in
tail suspension tests (Figure 5).
Figure 5 Effect of 100 mg/kg plant extract on
the immobility time of mice, plt0,05, n7
IV. DISCUSSION The decrease in vertical and
horizontal locomotor activities and exploratory
behavior indicates the reduce in CNS excitability
which might be closely related to sedation based
on depression of the CNS. In spite of decreased
spontaneous locomotor activity in the activity
cage, changes on motor coordination in Rota-Rod
test was not significant suggesting that this
effect is unrelated with neuromuscular blockade.
Decrease in the immobility time in tail
suspension test pointed out the antidepressant
like activity of the plant extract. Further
studies are neccessary to explain the mechanisms
of this antidepressant like activity.

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III. RESULTS The effects of the extract (100
mg/kg) on exploratory behavior, depression level,
spontaneous locomotor activity and motor
coordination were investigated in mice.
Administration of the extract induced a
significant decrease in the total number of head
dipping of mice in hole-board tests (Figure 1).
Figure 1 Effect of 100 mg/kg plant extract on
the total number of head dipping behaviour,
plt0,05, n7