evaluation of cestrum nocturnum for its anti-inflammatory - Journal of ...

Journal of Herbal Medicine and Toxicology 4 (1) 113-117 (2010)
ISSN : 0973-4643 Original Article
EVALUATION OF CESTRUM NOCTURNUM FOR ITS ANTI-
INFLAMMATORY AND ANALGESIC POTENTIALITY
Mazumder Avijit 1 *, Bhatt A 2 ., Bonde V. A 2 ., Shaikh A 2 . and Mazumder Rupa 1 .
1 Noida Institute of Engineering & Technology, 19 Knowledge Park II, Institutional Area, Greater Noida-201306
2 School of Pharmacy and Technology Management, NMIMS University, Vile Parle (West), Mumbai-400053
*Corresponding author email:avijitmazum@yahoo.com; Phone: +91-09871773644
Received- 4 th Sept, 2009, Revised-20 th Sept 2009, Accepted-
Abstract: The study evaluates the anti-inflammatory and analgesic potentiality of the
methanol extract of the leaves of Cestrum nocturnum and to provide scientific
justification for its traditional uses. Anti-inflammatory activity was carried out using
carrageenin induced paw oedema in rats whereas the analgesic activity was carried
out using hot plate method and acetic acid induced method on mice. The results indicate
that the extract posses strong dose dependant analgesic and anti-inflammatory activity
compared to standard drug.
Keywords: Cestrum nocturnum, leaves, anti-pyretic, anti-inflammatory , analgesic
INTRODUCTION
According to the World Health Organization [1],
medicinal plants would be the best source to obtain
drugs. Thus, scientific research on the healing
properties and bioactivity of natural products
especially of plant origin has been extensive
particularly in the western world. However, a rich
heritage of floral biodiversity is found in developing
countries . Cestrum nocturnum is an evergreen
woody shrub growing up to 4 metres high, commonly
known as Raat ki Rani (Queen of the night). It is a
popular landscape plant in warm climates. C.
nocturnum contains volatile oils [2-6], saponins,
lignans, glycosides and alkaloids[7,8]. Yuccagenin [9],
tigogenin, nocturnoside A & B, cesternoside A & B
[10-13] are some of its important constituents. The
leaves of C. nocturnum are mainly used as antiinflammatory,
antibacterial, antiepileptic, anti-malarial,
cardio-active, insect attractant, anticancer and
antifungal agent in herbal medicines. The claim for
the cardio tonic [14], anti-cancer [15-17], insecticidal
[18-20] and anti-epileptic [21] activities have been
proved. However there is no experimental evidence
of anti-inflammatory and analgesic activity of
C.nocturnum. Hence considering the above facts,
the present study was undertaken to screen the antiinflammatory
and analgesic potentiality of
C.nocturnum in experimental animal models.
MATERIALS AND METHOD
Collection and Preparation of extract
The leaves of Cestrum nocturnum Linn. were
collected from Nashik, Maharastra, India, identified
and authenticated by Dr. U. C. Bapat (Director,
Blatter Herbarium), St. Xavier’s College, Mumbai,
India. The leaves were shade dried, powdered and
pulverised. The powders so obtained were then passed
through sieve no. 85, weighed and stored in tightly
closed container. Then 70 gms of coarsely powdered
leaves of Cestrum nocturnum were subjected to
soxhlet extraction with 1 litre of methanol for 72 hours.
The resulting extract was concentrated under reduced
pressure using rotary vacuum evaporator at 40-50
°C to get the semisolid mass. The concentrated extract
was stored in a dessicator for further studies.
Phytochemical Testing
The extract so obtained was tested for the presence
113

Journal of Herbal Medicine & Toxicology
of various phytoconstituents with the help of different
chemical tests by standard methods [22,23].
Animals
Colony bred healthy rats of Wistar strain and albino
mice procured from School of Pharmacy and
Technology Management, Mumbai were used for the
114
study. They were housed in standard polypropylene
cages under room temperature (24 ± 2 O C); relative
humidity (60 % – 70 %) and exposed to 12:12hours
light:dark cycle. The rats were fed Nutrilab Rodent
Feed and drinking water filtered through an Aquaguard
water filter system ad libitum. They were allowed
to acclimatize for 5 days prior to commencement of
Table 1: Anti-inflammatory activity of leaves of Cestrum nocturnum in albino rats
Groups Dose
Control 10 ml/kg
Phenyl
butazone
Methanolic
extract of
Cestrum
nocturnum
100mg/kg
Low - 15
mg/kg
Medium - 20
mg/kg
High - 30
mg/kg
Paw volume response in millilitres at different time interval in Mean± SD
30 min 1hr 2hrs 3hrs 4hrs 6hrs
1.38±
0.138
1.30±
0.070**
1.36±
0.079
1.34±
0.046*
1.32±
0.031**
1.42±
0.170
1.34±
0.110**
1.39±
0.064
1.36±
0.057*
1.34±
0.032**
Values are expressed as mean ± SD; n=6 in each group,
1.45±
0.225
1.37±
0.14**
1.42±
0.065
1.38±
0.040*
1.36±
0.029**
1.48±
0.084
1.40±
0.077**
1.45±
0.057
1.41±
0.041*
1.39±
0.02**
1.51±
0.229
1.43±
0.032**
1.48±
0.055
1.44±
0.037*
1.43±
0.023**
1.54±
0.189
1.40±
0.035**
1.46±
0.055
1.42±
0.043*
1.40±
0.028**
**P < 0.001, *P < 0.01, Compared to control. Data was analyses by one way ANOVA followed by Duneet’s test
Table 2 : Analgesic activity of methanol extract of leaves of C. nocturnum by hot plate method:
Groups Dose
Control 10 ml/kg
Aspirin 200mg/kg
Methanolic
extract of
Cestrum
nocturnum
Low - 15
mg/kg
Medium -
20 mg/kg
High - 30
mg/kg
Paw Licking response in Seconds at different time intervals in Mean± SD
0 min. 30 min. 60 min. 90 min.
3.048 ±
0.041
3.768 ±
0.022**
(19.1%)
3.01±
0.014
(-1.26%)
3.115 ±
0.026*
(2.15%)
3.413 ±
0.029**
(10.69%)
4.173 ±
0.022
6.658 ±
0.043**
(37.3%)
4.21
± 0.018
(0.87%)
5.223 ±
0.022**
(20.1%)
6.263 ±
0.022**
(33.3%)
3.543 ±
0.030
8.568 ±
0.029**
(58.6%)
4.508 ±
0.009**
(21.4%)
6.113 ±
0.029**
(42.0%)
7.835 ±
0.036**
(54.7%)
2.895 ±
0.020
11
± 0.106**
(73.6%)
4.613 ±
0.049**
(37.2%)
6.363 ±
0.062**
(54.5%)
8.912 ±
0.018**
(67.5%)
(Figures in bracket indicates % pain inhibition) Values are expressed as mean ± SD; n=6 in each group,
*P < 0.01, **P < 0.001, Compared to control.
Data was analyses by one way ANOVA followed by Duneet’s test.

dosing. The protocol was ethically approved by IAEC
of the institute.
Drugs
Aspirin was used as a standard drug for comparing
the analgesic effect of the methanolic extract of C.
nocturnum. Phenyl butazone was used as a standard
drug for comparing the anti-inflammatory effect.
Anti-inflammatory Activity
Wister male/female albino rats (150-200 gm) of either
sex fasted overnight were weighed and segregated
into five groups.
Group 1- Carrageenan (0.05ml 1% solution)
Group 2- Phenyl butazone as Reference Standard
(100 mg/kg)
Group 3- Alcoholic Extract of Cestrum
nocturnum (15 mg/kg)
Group 4- Alcoholic Extract of Cestrum
nocturnum (20 mg/kg)
Group 5- Alcoholic Extract of Cestrum
nocturnum (30 mg/kg).
A ring was marked on the left paw of each animal so
that constant length of paw could be dipped each time.
The initial paw volume (of left leg) of each rat was
measured using plethysmometer. One hour after oral
administration of aqueous suspension of phenyl
butazone and test drug all the rats were challenged
by an injection of 0.05 ml of 1% w/v solution of
carrageenan into the planter side of the left hind paw.
The paw volume was measured by using
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Mazumder et al.
Table 3 : Analgesic activity of methanol extract of leaves of Cestrum Nocturnum by acetic acid
induced writhing method:
Groups Dose Number of writhings (10 min.) Mean ± SD) % inhibition
Control 10 ml/kg 34 ± 1.3 --
Aspirin 200mg/kg 10 ± 0.96** 70.5 %
Methanolic extract of
C. nocturnum
Low - 15 mg/kg 31 ± 2.4 8.8 %
Medium - 20 mg/kg 29 ± 3.7 14.7 %
High - 30 mg/kg 15 ± 2.6** 55.8 %
Values are expressed as mean ± SD; n=6 in each group, **P < 0.001 Compared to control.
Data was analyses by ANOVA followed by "Bonferroni's Multiple Comparison Test".
plethysmometer immediately after injection, 30min,
1hr, 2hrs, 3hrs, 4hrs, 6hrs and 24hrs after challenge
with carrageenan and the percent increase in the paw
volume before and after sub plantar injection of
carrageenan was calculated. [24,25].
Analgesic activity
Analgesic activity was carried out by two methods,
a) Hot Plate Method
b) Acetic Acid induced Method
Hot Plate Method
In this method albino mice (20-40 gm.) were
segregated into five groups.
Group 1- Distilled Water
Group 2- Aspirin as Standard (200 mg/kg)
Group 3- Alcoholic Extract of Cestrum
Nocturnum (15 mg/kg)
Group 4- Alcoholic Extract of Cestrum
Nocturnum (20 mg/kg)
Group 5- Alcoholic Extract of Cestrum
Nocturnum (30 mg/kg)
The pain threshold (Number of licking of paw/jumping)
were measured at 30, 60, 90 minute after oral
administration of standard and test solution by the use
of hot plate maintained at 55 0 C. A cut off time of 15
second was taken as maximum analgesic response
to avoid injury to the paws. The percent increase in
reaction time at each time interval was calculated as
percent inhibition [26-28].

Journal of Herbal Medicine & Toxicology
Acetic Acid induced method
The extract at doses ranging from 30 to 15 mg/kg
was orally administered to mice 1 hour before i.p.
injection of 0.6% (v/v) acetic acid, at a dose of 10 ml/
kg. Distilled water was used as a control treatment
while the reference group received 200 mg/kg of
acetyl salicylic acid (ASA) as a standard. The
writhings (a syndrome characterized by a wave of
contraction of the abdominal musculature followed
by extension of hind limbs) that occurred between 5
and 15 min after acetic acid were counted. A reduction
in the writhing number as compared to the control
group was considered as evidence for the analgesia,
which was expressed as percent inhibition of writhings
[29].
RESULTS AND DISCUSSIONS
The extract showed a dose dependant change in paw
volume. However 30 mg/kg of the methanolic extract
of C. nocturnum exhibited a significant decrease in
paw volume after 6 hrs comparable to the standard
reference drug phenyl butazone (100mg/kg). (Table-
1). However, the methanolic extract of C. nocturnum
leaves showed a significant and dose-dependent
analgesic activity in hot plate method and acetic acid
induced method used. The activities found at a dose
of 30 mg/kg was comparable to the reference drug
aspirin at 200 mg/kg by both the methods used (Tables
2 and 3). It was reported that the analgesic action of
Cleome viscose is attributed to nicotine [27] which is
also interestingly found to be one of the phytochemical
present in the leaves of our tested plant. Further, the
anti-inflammatory potentiality of 3-[(2S)-1methylpyrrolidin-2-yl]
pyridine was demonstrated in
Anacardium occidentale [25]. Thus the antiinflammatory
action of our screened C. nocturnum
can be claimed to be due to 3-[(2S)-1methylpyrrolidin-2-yl]
pyridine present in the leaves
of our screened plant.
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