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Archives of Medicine and Health Sciences / Jan-Jun 2014 / Vol 2 | Issue 1 5
Original Article
Introduction
Anxiety is a feeling of nervousness, an unpleasant emotion
due to the anticipation of unreal or threat imagined. The
complexity of life in modern society frequently leads
to varying degree of anxiety. One-eighth of the world
population is suffering from anxiety.[1] Benzodiazepines
(bzds), barbiturates, tricyclic antidepressants (TCAs) have
been used for the treatment of anxiety disorders.[2] The
side-effects of these drugs are sedation, rebound insomnia,
withdrawal and tolerance, sexual dysfunction, muscle
relaxation etc.[1] Therefore, an agent with good therapeutic
Shankar K. Bhat, Anu Elizabeth Joy
Department of Physiology, Yenepoya Medical College, Mangalore, Karnataka, India
Antianxiety effect of ethanolic extract of leaves of
Moringa oleifera
in Swiss albino mice
ABSTRACT
Background: Anxiety disorder is one of the most common mental ailments exhibited by humans. It can cause considerable distress and
debility. Anxiety is portrayed as a frame of mind concerned about future in association with preparation for possible, upcoming undesirable
happenings. The present treatment for the disorder is having a lot of side-effects. An agent with good therapeutic effect and less side-effects is
needed for the treatment of anxiety. Objectives: To investigate the anxiolytic activity of ethanolic extract of Moringa oleifera leaves in Swiss
albino mice. Materials and Methods: The ethanolic extract of leaves of Moringa oleifera (200 mg/kg, i.p) was studied for its anxiolytic effect
on Swiss albino mice by using Elevated Plus Maze (EPM) and Light Dark Arena (LDA) test. Results: The ethanolic extract of the leaves of
Moringa oleifera (200 mg/kg, i.p) demonstrated signifi cant (P < 0.001) anxiolytic activity in EPM and LDA models of anxiety. Conclusion:
The data suggests that the ethanolic extract of Moringa oleifera leaves may have produced its anxiolytic effects via multiple mechanisms.
Key words: Anxiety, swiss albino mice, moringa oleifera, ethanolic extract, anxiolytic activity
effect and less side-effects is needed for the treatment of
anxiety.
Moringa oleifera commonly known as drumstick tree in
English; Subhanjana in Sanskrit; Saguna in Hindi; Sigru
in Malayalam;[3] Nugga in Kannada; and Murungai in Tamil
belongs to the family Moringaceae.[4] It’s a soft wooded tree,
and all the parts of the tree are edible. Traditionally, the leaves
are used for the treatment of variety of disorder. Leaves
are known to have anti-hyperglycemic, hepatoprotective,
anti-hyperlipidemic, anti-microbial, anti-inflammatory, anti-
convulsant, antioxidant properties.[5]
It is a natural anthelmintic, antibiotic, detoxifier, outstanding
immune builder and is used in many countries to treat
malnutrition and malaria. It is used in water purification and,
therefore, helps in reducing the incidence of water-borne
diseases.[6] Its a known fact that oxidative stress also plays an
important role in the etiology of anxiety disorders.[7] This study is
to investigate the effect of ethanolic extract of Moringa oleifera
leaves in animal models of anxiety.
Corresponding Author:
Ms. Anu Elizabeth Joy, Department of Physiology, Yenepoya Medical College, Yenepoya University, Deralakatte, Mangalore - 575 018, Karnataka, India.
E-mail: joyliz07@yahoo.co.in
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DOI:
10.4103/2321-4848.133771
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Bhat and Joy: Antianxiety effect of Moringa oleifera
6 Archives of Medicine and Health Sciences / Jan-Jun 2014 / Vol 2 | Issue 1
Materials and Methods
Animals
Young adult Swiss albino mice of either sex weighing 25-
30 g were used in this study after obtaining Institutional
Animal Ethical Committee Clearance. The mice were
maintained under standard conditions in the animal house.
The mice were kept in polypropylene cages and maintained
on standard pellet diet and water ad libitum. Animals were
acclimatized under standard laboratory condition and were
kept in 12 hr day and night cycle for 7 days before conducting
experiments.
Drugs / Dose / Route of administration
Diazepam (Cipla Ltd.) was obtained from Yenepoya Hospital
Pharmacy in Mangalore. It was administered at a dose of
(1 mg/ kg i.p).
Instruments
Soxhlet apparatus was used to prepare the plant extract.
Elevated plus Maze apparatus and Light Dark Arena
apparatus were used for screening the effect on anxiety.
Plant materials
Leaves of Moringa oleifera were used for the study. The
fresh leaves were collected from Thiruvalla, Kerala.
They were shade-dried. The dry leaves were grinded into
coarse powder and extracted using Soxhlet apparatus. The
leaves were authentified by Dr. Noeline. J. Pinto, Head of
the Department, Botany, St. Agnes College, Mangalore,
Karnataka, India.
Preparation of the extracts
Moringa oleifera ethanolic extract [MOEE]:
A weighed quantity (500 g) of the coarse powder was taken
and extracted with ethanol (90%) in a Soxhlet apparatus. The
extract was concentrated on a water bath at a temperature
not exceeding 60ºC. The ethanolic extract was suspended
in distilled water. MOEE was administered at a dose of
200 mg/kg/bodyweight i.p.
Experimental design
Thirty-six animals were used in this study. The animals were
divided into 3 groups. Each group consisting of 6 males and
6 females (n = 12).
Group-I: Normal Saline (0.1 ml i.p)
Group-II: MOEE (200 mg/kg i.p)
Group-III: Diazepam (1 mg/ kg i.p)
After an hour of the administration of test compounds, the
animals were taken for the following tests for screening its
anxiolytic activity.
Elevated plus maze [EPM].[1,2,8]
To measure the level of anxiety in rodents, elevated plus
maze was used. Three potential anxiogenic factors are
open space, height, and novelty. The cross-shaped maze
consists of 4 arms that are interconnected by a central
platform. Two opposing arms are surrounded by side and
end-walls (closed arms), whereas the remaining 2 arms
are unprotected (open arms). The set-up consists of a
maze of 2 open arms (16 cm × 5 cm), crossed with walls
(12 cm high) and central platform (5 cm × 5 cm). The maze
is suspended 25 cm above the room floor. The animal was
placed on the central platform, facing one of the enclosed
arms, and observed for 5 minutes (300 seconds). During
the 5-min test period, the time spent in open and enclosed
arms were recorded
Light dark arena (LDA).[9]
The instrument consist of 2 parts, 1/3 with opaque walls
and a cover (dark compartment), whereas the remaining
2/3 was open and illuminated (light compartment). The door
between the two compartments permits mouse to move
from one side to another. Each mouse was released in the
light compartment and observed for 5 minutes. Time spent
in light and dark compartment were recorded.
Statistical analysis
Results were expressed as mean ± SD. One-way analysis
of variance (ANOVA) was carried out, and the statistical
comparisons among the groups were performed with
Tukey Krammer test using a statistical package program.
P value < 0.001 was considered as significant.
Results
Elevated plus maze
MOEE-treated animals (Group II) and Diazepam-treated
animals (Group III) showed a significant (P < 0.001) increase in
the time spent in open arms [Table 1] by EPM test on comparing
with the normal (Group I). There was significant difference in the
time spent in the open arm between the MOEE-treated animals
(Group II) and Diazepam-treated ones (Group III).
Light dark arena
MOEE-treated animals (Group II) and Diazepam-treated anim
als (Group III) showed a significant (P < 0.001) increase in
the time spent in bright arena and dark arena [Table 2] by
LDA test on comparing with the normal (Group I). There is
no significant difference between the MOEE-treated animals
(Group II) and Diazepam-treated ones (Group III).
The above observations suggest that Moringa oleifera has
anxiolytic activity.
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Bhat and Joy: Antianxiety effect of Moringa oleifera
Archives of Medicine and Health Sciences / Jan-Jun 2014 / Vol 2 | Issue 1 7
Discussion
Anxiety is a frame of mind concerned about future in
association with preparation for possible, upcoming
undesirable happenings.[10] Neurotransmitters play an
important role in the pathophysiology of anxiety disorders.[11]
It is a well-known fact that low level of Gamma Amino Butyric
Acid (GABA) in CNS is most commonly linked with anxiety
disorders.[12]
Serotonin (5-HT) has also an important role in the progress of
anxiety disorders. Studies showed that patients with anxiety
ailments have genetic polymorphisms in the 5-HT transporter.[13]
Apart from GABA and 5-HT, Dopamine and Norepinephrine
(NE) also have an important role in the progress of anxiety
disorders. Several animal studies including rodents and
monkeys have suggested that reduced dopamine activity
is associated with augmented anxiety. Several pre-clinical
studies showed that there is an increased synthesis, release,
and turnover of Norepinephrine in anxiety and stress, thus
suggesting a role of NE in anxiety. This is further confirmed
by the fact that agents which reduce the activity of NE in
CNS have shown anti-anxiety effect.[14,15]
Oxidative stress also plays an important role in the etiology
of anxiety disorders. Anxiety disorders can also be due to
free radical-induced damage to neurotransmitter system.[13,16]
The results have shown that Moringa oleifera has anxiolytic
property. Relatively very few information exist on the anxiolytic
property of Moringa oleifera. The anxiolytic property can be
due to its modulating role on the above neurotransmitters or
due to its antioxidant property. Further studies are ongoing
to find the active phytoconstituents present in the indigenous
medicinal plant responsible for this anxiolytic property.
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Table 1: Showing anxiolytic effect of MOEE by elevated plus maze test
Groups Drugs Time spend in each arm in seconds
Open Closed
I Normal 9.16±1.84 258.33±8.08
II MO 59.33±1.8a,b 182.66±11.5 a,c
III Diazepam 50.16±1.69a193.11±15.6 a
One-Way ANOVA, followed by Tukey Kramer multiple comparison test.
aP < 0.001 → extremely signifi cant. (Comparing group II and III with I),
bP < 0.001 → extremely signifi cant. (Comparing group III with II in open arm),
cP > 0.05 → not signifi cant. (Comparing group III with II in closed arm), n = 12,
MO: Moringa Oleifera
Table 2: Showing anxiolytic effect of MOEE by light dark arena test
Group Drugs Time spend in each arm in seconds
Bright Dark
I Normal 34±1.7 252.16±14.8
II MO 108.33±4.4 a,b 169.8±14.1 a,b
III Diazepam 109.9±8.9 a 166.9±10.7 a
One-Way ANOVA, followed by Tukey Kramer multiple comparison test. aP < 0.001
→ extremely signifi cant. (Comparing group II and III with I), bP < 0.05 → not
signifi cant. (Comparing group III with II), n = 12, MO: Moringa Oleifera
How to cite this article: Bhat SK, Joy AE. Antianxiety effect of ethanolic
extract of leaves of Moringa oleifera in Swiss albino mice. Arch Med Health
Sci 2014;2:5-7.
Source of Support: Nil, Confl ict of Interest: None declared.
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