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74 Original article

Antipyretic effect of ethanolic extract of Moringa oleifera leaves

on albino rats
Ayon Bhattacharyaa, Rasmirekha Beheraa, Divya Agrawalb, Pratap K. Sahuc,
Sanjay Kumara, Sudhanshu S. Mishraa

Departments of aPharmacology, bAnatomy, Objective

IMS and SUM Hospital, cDepartment of
Pharmacology, SPS, Siksha O Anusandhan
(SOA) University, Bhubaneswar, Odisha, India
Correspondence to Ayon Bhattacharya, MD,
Department of Pharmacology, IMS & SUM
Hospital, Kalinga Nagar, B.O. Ghatikia,
Bhubaneswar 751003, Odisha, India
Tel: +91 923 808 4500; Fax: 00916742386293;
e-mail: ayonbhattacharya@yahoo.in
Received 11 April 2014
Accepted 18 May 2014
Tanta Medical Journal 2014, 42(2):74–78
The aim of the study was to evaluate the antipyretic activity of ethanolic leaf extract of Moringa
oleifera using Brewer’s yeast-induced pyrexia model.
Materials and methods
It was a randomized controlled experimental study. A total of 60 rats were taken, dividing them
in six groups, each containing 10 rats. Ethanolic extract of M. oleifera (EMO) was administered
at 50, 100, 200, and 400 mg/kg doses orally to the respective four groups. The control group
was fed with normal saline at 2 ml/kg. A 20% suspension of Brewer’s yeast in normal saline
was injected subcutaneously at a dose of 10 ml/kg body weight below the nape of neck of rats
in all groups. Pyrexia developed after 10 h of Brewer’s yeast injection and the temperature was
recorded. Animals which showed a rise in body temperature to at least 39°C were included in
the study, allowing a minimal of six rats in each group, total of 36 rats. Drugs were given after
development of pyrexia and temperatures were recorded. Paracetamol at 100 ml/kg orally
was taken as the standard drug.
Results
The ethanolic leaf extract of M. oleifera showed significant (P < 0.05) antipyretic activity at
100, 200, and 400 mg/kg. Paracetamol showed significant antipyretic activity from 15 min of
drug administration to 12 h. EMO at a dose of 50 mg/kg did not show antipyretic effect. The
onset of action of EMO 100 mg/kg was found to be 2 h and that of 200 and 400 mg/kg was
found to be 30 min. For all the doses, the antipyretic effect lasted up to 12 h.
Conclusion
The ethanolic leaf extract of M. oleifera exhibited significant (P < 0.05) antipyretic activity at
100, 200, and 400 mg/kg.

Keywords:
antipyretic, Brewer’s yeast, Moringa oleifera

Tanta Medical Journal 42(2):74–78

© 2014 Tanta Medical Journal
1110-1415

Arabia, is now cultivated in Philippines, Cambodia,

Introduction
Herbal medicines are assuming great importance in the
primary healthcare of individual and society. Over 75% of
the world population relies on plant extracts for healthcare,
and more than 30% of the entire plant species at one time
or the other are used for medicinal purposes [1]. The
WHO encourages the use of herbal remedies that have
proved efficacious in primary healthcare [2].
Pyrexia or fever is defined as the elevation of body
temperature. It is a natural defense mechanism or
response to tissue damage, inflammation, malignancy,
or graft rejection. Because of poor hygiene practices
and malnutrition, children from developing countries
suffer from various infections, which present as fever.
These fevers are accompanied by personal discomforts
such as pain (myalgia), which undoubtedly leads to
morbidity and mortality [3].
Moringa oleifera, native of the western and sub-
Himalayan tracts, India, Pakistan, Asia, Africa, and
1110-1415 © 2014 Tanta Medical Journal
Central America, North and South America, and
Caribbean islands [4]. It is a small, fast-growing
evergreen or deciduous tree that grows up to 10–12 m in
height, with open crown of drooping fragile branches,
feathery foliage of trip innate leaves, and thick corky,
whitish bark [5]. This plant has been known since the
ancient Egyptian and Roman times for the multitude
of medicinal and other uses. In the Charaka Samhita,
it has been stated to be used for various infections of
ear, nose, and throat. M. oleifera is known by various
names such as horseradish tree, drumstick tree, ben oil
tree, miracle tree, and ‘Mother’s best friend’ [6]. The
leaves are of high nutritive value and contain a rare
combination of ascorbic acid, carotenes, flavonoids,
isoquercetin, and glycosides such as niazirin,
4-hydroxymellein, β-sitosterol, vanillin, caffeoylquinic,
kaempferol, and zeatin [7,8]. As a result of the anti-
inflammatory action of these bioactive constituents,
the antipyretic action can be hypothesized. Antipyretic
potential of the ethanolic seed extract of M. oleifera
has been studied [9] but not that of leaves. Hence, the
DOI: 10.4103/1110-1415.137810
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Antipyretic effect of Moringa oleifera leaves on albino rats Bhattacharya et al.
present study was undertaken to study the antipyretic
potential of leaf extract of M. oleifera.
Materials and methods
The study protocol was approved by Institutional
Animal Ethical Committee (IAEC), Siksha O
Anusandhan (SOA) University. Experiments were
performed in Department of Pharmacology, IMS &
SUM Hospital, Bhubaneswar. It was a randomized
controlled study.
Collection of plant material
The leaves were collected from the local areas of
Syampur (Bhubaneswar, Odisha), and its identity
was confirmed taxonomically by Dr P.C. Panda,
scientist of Regional Plant Research Centre (RPRC),
Bhubaneswar.
Extract preparation
The leaves (500 g) of M. oleifera were obtained locally
and shade dried and powdered. It was extracted with
90% ethanol in a Soxhlet apparatus for 8 h. Extract
was filtered (Whitman filter paper no. 1), concentrated
in a rotary evaporator to yield a semisolid mass of
42 g (8.4% w/w), stored at 4°C, and used for oral
administration.
Animals
The animals were procured from central animal house
IMS & SUM Hospital. Wistar albino rats of either
sex (100–200 g) were used. Food and water were given
ad libitum. Animals were acclimatized to laboratory
conditions for 7 days before the experiments. The
study was approved by the IAEC of SOA University,
Bhubaneswar, under the approval number 22/12/
IAEC/SPS/SOA. All experiments and animal care
were according to the CPCSEA and good laboratory
practice guidelines. No animals were killed at the end
of the study.
Chemicals
Paracetamol (Dr Reddy’s Laboratory, Hyderabad,
India), yeast extract powder (HiMedia Laboratories
Pvt Ltd, Mumbai, India), and other solvents were of
analytical grade.
Brewer’s yeast-induced pyrexia
The animals were randomly divided into six groups,
each group consisting of 10 rats; a total of 60 rats were
used in the study by randomized sampling technique:
group I (control, normal saline given orally at
75
2 ml/kg body weight); group II (standard, paracetamol
100 mg/kg); groups III, IV, V, and VI [ethanolic extract
of M. oleifera (EMO) 50, 100, 200, and 400 mg/kg,
respectively]. This is regarded as a classical method for
antipyresis testing. Wistar strain of albino rats of either
sex weighing 100–200 g were used for the study. The
animals were fasted for 18 h before commencement of
the experiment, but water was provided ad libitum. An
initial rectal temperature was recorded using a rectal
thermometer to a depth of 1.5 cm in the rectum of
rats. Animals with a body temperature between 36
and 38°C were included in the study. A 20% Brewer’s
yeast in 0.9% w/v saline was injected subcutaneously
below the nape of neck at a dose of 10 ml/kg thereafter.
The injection site was massaged to ensure the spread
of suspension below the skin. Room temperature was
maintained at 22–24°C. After the yeast injection, food
was immediately withdrawn. After 10 h postchallenge,
the rise in rectal temperature was recorded. Animals
which showed a rise in body temperature to at least
39°C were included in the study, allowing a minimal
of six rats in each group, total of 36 rats. The animals
received the standard (paracetamol 100 mg/kg) or
the test compound (EMO 50, 100, 200, 400 mg/kg)
by oral administration, and the rectal temperature
was recorded at 0, 0.25, 0.50, 1, 2, 3, 4, 5, 6, 12, and
24 h after dosing. The maximum reduction in average
rectal temperature in comparison with the control
hyperpyrexia group was calculated and compared [10].
Results
Subcutaneous injection of the pyrogenic dose of yeast
produced elevated changes in rectal temperature, which
is shown in Table 1. The EMO showed a significant
(P < 0.05) decrease in rectal temperature at doses 100,
Figure 1
Line graph showing the effect of paracetamol and ethanolic extract
of Moringa oleifera (EMO) on yeast-induced pyrexia.
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76 Tanta Medical Journal

200, and 400 mg/kg when compared with the control.

37.00 ± 0.37
37.1 ± 0.27*** 37.08 ± 0.24*** 37.08 ± 0.31

37.16 ± 0.23
37.1 ± 0.23
37.5 ± 0.50** 37.03 ± 0.48
39.25 ± 0.16 39.08 ± 0.33 38.83 ± 0.33* 38.56 ± 0.35** 38.15 ± 0.37***37.8 ± 0.44*** 37.65 ± 0.52***37.38 ± 0.64***37.43 ± 0.48*** 37.33 ± 0.56** 37.18 ± 0.23
However, no significant decrease in mean temperature

24 h
was noted by EMO 50 mg/kg when compared with
control throughout the study period (Table 1 and Fig. 1).
EMO at doses of 50, 100, 200, and 400 mg/kg showed

37.61 ± 0.26*
38.21 ± 0.27

38.15 ± 0.22
12 h a progressive decline in mean temperature pattern with
the increase in the dose (Fig. 2). Paracetamol showed
significant (P < 0.05) decrease in rectal temperature
from 0.25 to 12 h. The onset of action of paracetamol

37.9 ± 0.43**
38.66 ± 0.15* 38.61 ± 0.19** 38.41 ± 0.27** 38.23 ± 0.20* 38.23 ± 0.21*
38.81 ± 0.18
38.8 ± 0.26

was 15 min and the body temperature became normal

6h
Mean rectal temperature recordings after 10 h of Brewer’s yeast injection in degree centigrade

after 12 h. EMO at doses 100 mg/kg showed its onset

of action from 2 h and EMO 200 and 400 mg/kg from
30 min (Fig. 2). The line graph (Fig. 3) shows that
37.18 ± 0.26 39.15 ± 0.18 39.01 ± 0.33 38.85 ± 0.37* 38.73 ± 0.54* 38.48 ± 0.48** 38.25 ± 0.42***38.15 ± 0.49***37.95 ± 0.51**
37.8 ± 0.62***37.06 ± 0.38*** 37.1 ± 0.28***37.13 ± 0.27***37.1 ± 0.35***

paracetamol registered a phenomenal decrease in mean

38.95 ± 0.34

38.91 ± 0.32

temperature between 15 min and 2 h, and thereafter

5h

maintaining a steady mean temperature. Figure 4

illustrates that the most significant (P < 0.05) decrease
in round-the-clock mean temperature in this study was
39.23 ± 0.41

38.93 ± 0.13

shown by paracetamol followed by EMO at 400 mg/kg.

4h

Statistical analysis was performed using one-way analysis

of variance followed by post-hoc Dunnette’s test. The
results were recorded as mean ± SD.
39.31 ± 0.44

39.05 ± 0.20
3h

Discussion
BBT, basal body temperature; EMO, ethanolic extract of Moringa oleifera; *P < 0.05; **P < 0.01; ***P < 0.001.

Brewer’s yeast (lipopolysaccharide, which is the cell wall

39.33 ± 0.45

39.16 ± 0.12
Table 1 Effect of paracetamol and ethanolic extract of Moringa oleifera on yeast-induced pyrexia

component of Gram negative bacteria) is an exogenous

2h

pyrogen that binds to the immunological protein called

the lipopolysaccharide binding protein [11]. This
binding results in the synthesis and release of various
39.41 ± 0.33

39.13 ± 0.16
38.95 ± 0.13

endogenous cytokine factors such as interleukin (IL)-1,

1h

IL-6, and TNFa, which activate the arachidonic acid

pathway, and ultimately result in the synthesis and
release of prostaglandin E2 (PGE2) [12]. Yeast-induced
39.2 ± 0.16 38.75 ± 0.30* 38.46 ± 0.39**
37.16 ± 0.17 39.18 ± 0.13 39.15 ± 0.17 39.28 ± 0.30

39.2 ± 0.30 39.25 ± 0.19

37.08 ± 0.18 39.23 ± 0.12 39.13 ± 0.19 39.05 ± 0.15

pyrexia is called as pathogenic fever [13].

0.50 h

According to the classical view, fever is induced by

inflammatory mediators (IL-1, IL-2, TNFα, others)
released by the peripheral mononuclear macrophages
0.25 h

and other immune cells [14,15]. These fever-promoting

cytokines are transported from blood to the brain
by specific carriers [16]. Cytokines are transported
37.28 ± 0.21 39.28 ± 0.19

by bloodstream and enter the brain through the

0h

circumventricular organs [17]. Alternatively, the

cytokines could interact with their receptors on brain
endothelial cells [18] or perivascular tissue [19]. This
37.16 ± 0.34

37.16 ± 0.12

assumed mechanism of fever induction is known as

BBT

the humoral hypothesis of fever induction. These

proinflammatory mediators act on the preoptic/
anterior hypothalamus triggering the release of PGE2
(paracetamol 100

EMO 100 mg/kg

EMO 200 mg/kg
EMO 400 mg/kg

produced from cyclooxygenase (COX)-2, and thus

EMO 50 mg/kg

elevating the body temperature [20].

Standard
Control

mg/kg)

An effective febrifuge such as paracetamol acts

???

by blocking the effect of these pyrogens in the

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Antipyretic effect of Moringa oleifera leaves on albino rats Bhattacharya et al.
Figure 2
Line graph showing the effect of ethanolic extract of Moringa oleifera
(EMO) 100, 200, and 400 mg/kg in comparison with control on yeast-
induced pyrexia model. *P < 0.05, **P < 0.01, ***P < 0.001.
Figure 3
Line graph showing the effect of ethanolic extract of Moringa oleifera
(EMO) 400 mg/kg and paracetamol on yeast-induced pyrexia.
*P < 0.05, **P < 0.01, ***P < 0.001.
Figure 4
Radar diagram showing the effect of ethanolic extract of Moringa
oleifera (EMO) and paracetamol on yeast-induced pyrexia model.
77
temperature-sensitive neurons in the preoptic region
of the hypothalamus to COX formation of PGE2 [21].
However, in inflammatory lesions, paracetamol is a
poor inhibitor of COX due to presence of peroxides.
The anti-inflammatory activity of M. oleifera is
attributed to its COX inhibitory activity [22]. Thus,
the present study postulates that EMO could reduce
pyrexia by reducing the concentration of PGE2 in
the hypothalamus or by interrupting the steps that
connect the peripheral inflammation with the central
production of PGE2 or both [23,24].
The phytochemical ingredients in the leaf extract, such
as phenolics, flavonoids, tannins, saponins, terpenoids,
isoquercetin, and glycosides such as niazirin,
4-hydroxymellein, β-sitosterol, and vanillin, could be
responsible for its antipyretic activity [6,25].
Conclusion
Therefore, from the results of the study, we can deduce
that EMO could be used as an antipyretic agent at
100, 200, and 400 mg/kg. However, further studies are
necessary to examine the mechanism of action of the
antipyretic activity and to isolate the active compounds
responsible for this pharmacological activity.
Acknowledgements
Conflicts of interest
There are no conflicts of interest.
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