Screening of Antipyretic and Analgesic
Potential of Ethanol Extract of Cassytha filiformis Leaves
Ram Kumar Sahu1*, Amit Roy1, SaurabhKothiya2, Anup Kumar
Maurya3, Rajesh Kumar4
1Columbia Institute of Pharmacy, Tekari,
Raipur (C.G.)-493111, India.
2Oriental
College of Pharmacy, Raisen Road, Bhopal-462021
(M.P.), India.
3Shridhar University, Pilani-333031 (Rajasthan), India.
4Institute of Pharmacy, Kathog-177101 (H.P.), India
ABSTRACT:
The present study was
undertaken to evaluate the antipyretic and analgesic activities of the alcohol
extract of Cassytha filiformis leaves.
The antipyretic activity of C. filiformis (25mg/kg, 50mg/kg and 100mg/kg) was
studied in Brewer’s yeast induced pyrexia in rats. The
analgesic activity of C. filiformis (25mg/kg, 50mg/kg and 100mg/kg) was studied
using Eddy’s hot plate method and heat conduction method in mice. The extract
showed significant reduction in the elevated body temperature of rat which was
compared with standard Paracetamol. The extract produced significant increase
in the reaction time by Eddy”s hot plate method and
tail flick method in mice which was compared with standard diclofenac
sodium. The
alcohol extract of leaves of C. filiformis may have antipyretic and analgesic activities.
KEYWORDS: Cassytha filiformis, antipyretic, pyrexia,
analgesic, paracetamol
INTRODUCTION:
Cassytha filiformis L. is a
leafless, climbing, twining, vine-like, autoparasitic
and plant-hyperparasitic phanerogam
(seed-bearing plant) in the plant family Lauraceae1. The individual
stems that are copiously branched have a range of 1 to 3 millimeters in the
diameter and finally attain a maximum length of 10 to 20 feet. The flowers were
borne all over the year with a bisexual nature with a stamen and perianth arrangement that looks similar to the ordinary
avocado. The ovary is the one first getting exposed and later on becomes
enveloped by the enlargement and then the over growth of the calyx tube occurs.
The fruit is about the size of the large pea and gets closely enclosed by means
of the succulent calyx. The single seed possess a membranous testa and the separation between cotyledons is not properly
defined2.
This
plant is distributed throughout India and is used for medicinal purpose in
China, Indochina, Madagascar and South Africa. C. filiformis is used as antiplatelet agent, vesorelaxant,
alpha-adrenoreceptar antagonist and antitrypnosomal agent. Some of the isolated compound from this plant are aporphine
alkaloid, oxo-aporphine alkaloid, cassyformine,
filiformine, cathaformine, lignan, actinodophine, and octenine. In ayurveda, C. filiformis
is used as substitute for Cuscuta reflexa3.
In recent times, focus on plant research
has increased all over the world and a large body of evidence has been
collected to show immense potential of medicinal plants used in various
traditional systems. Medicinal herbs are highly highlighted due to their wide
use and less side effects. Therefore the
present study was undertaken to evaluate the antipyretic and analgesic
activities of the alcohol extract of C. filiformis leaves.
MATERIALS
AND METHODS:
Plant
material
The proposed study of C. filiformis were collected from the Van Vihar, Bhopal, Madhya Pradesh, with the help of local
people and field botanist.
Preparation of ethanol extract
The powder of leaves (300gm) of C. filiformis, was packed well in Soxhlet apparatus and extracted with ethanol until the
completion of the extraction. The extract was filtered while hot, and the
resultant extract was distilled in vacuum under reduced pressure in order to
remove the solvent completely and dried in a desiccators.
Animals
Male
wistar albino rats and mice were kept in quarantine
for 10 days under standard husbandry conditions (27.3o, Relative
humidity 65 ±10%) for 12 hrs in dark and light cycle respectively and were
given standard food and water ad. libitum.
Antipyretic studies
The
body temperature of each albino Wistar mice was recorded by measuring rectal
temperature at predetermined intervals. Albino wistar
mice were fasted overnight with water ad libitum before the experiments. Pyrexia was induced by
subcutaneously injecting 20% (W/V) brewer's yeast suspension (10 ml/kg) into
the animal's dorsum region. The rectal temperature of each rat was again
recorded after 24 h of yeast administration. Mice that did not show a minimum
increase of 0.5 ºC in temperature 24 h after yeast injection were
discarded. Thirty selected mice were grouped into five and immediately treated
as follows: group I received normal saline, group II received 10 mg/kg
paracetamol, while groups III, IV and V received ethanol extracts 25, 50 and
100 mg/kg respectively i.p. Rectal temperature of all
the mice was then recorded by inserting digital thermometer into the rectum of
each mice at thirty minutes4,5.
Analgesic activity
(i)
Hot plate method
The animals were divided
into five groups with six mice in each group. Group I animals served as
control, animals of Group II received Diclofenac sodium at 10 mg/kg body weight
while animals of Group III, Group IV and Group V were treated with 25, 50 and
100 mg/kg body weight (s.c.) of the ethanol extract.
The animals were placed on Eddy’s hot plate kept at a temperature of 55±0.5°C.
A cut off period of 15s was observed to avoid damage to the paw. Reaction time
was recorded when animals licked their fore or hind paws, or jumped prior to
and 0, 30, 60 and 90 min after administration of the samples.
(ii) Hot Tail Flick method
The animals were divided
into five groups with six mice in each group. Group I animals served as
control, animals of Group II received Diclofenac sodium at 10 mg/kg body weight
while animals of Group III, Group IV and Group V were treated with 25, 50 and
100 mg/kg body weight (s.c.) of the ethanol extract.
The animals were placed on Eddy’s hot plate kept at a temperature of 55±0.5°C.
A cut off period of 15s was observed to avoid damage to the paw. Reaction time
was recorded when animals licked their fore or hind paws, or jumped prior to
and 0, 30, 60 and 90 min after administration of the samples6,8.
Statistical Analysis
The
results are expressed as mean ± SEM of six independent experiments. Statistical
significance between group was evaluated by one-way analysis of variance
(ANOVA) followed by Dunnet’s test. A P < 0.05
value was considered as statistically significant.
RESULT AND DISCUSSION:
The ethanol extract of C. filiformis
leaves decreased the fever induced by yeast in rats. From the result, the ethanol
extracts (25 mg/kg, 50 mg/kg and 100 mg/kg) fever lowering effect comparable
with the standard drug, paracetamol, at 10 mg/ml concentration (table 1). Thus,
it is possible that active compound(s) for antipyretic action may be included
in the ethanol extract. The fever condition entails enhanced formation of
cytokines such as interleukins, interferons and tumor
necrosis factor, and the cytokines increase the synthesis of prostaglandin E2.
Paracetamol suppresses this response by inhibiting the synthesis of
prostaglandin E2. The extract may be involved in the inhibition of some of
these substances inducing fever.
The
result of hot plate test indicated a significant increase in reaction time at
0.5, 1 and 1.5 hours as comparable to the reference drug diclofenac
sodium (10 mg/kg; s.c.)
which is showed in table 2. The results obtained from hot tail flick
experiments are shown in table 3, in this model, administration of ethanol
extract (25 mg/kg, 50 mg/kg
and 100 mg/kg)
showed significant protection against the pain induction.
The
result from hot and cold tail flick test also gave additional evidences for the
analgesic activity of the extract. The activity may be attributed due to the
presence of flavonoids, alkaloids and other bioactive
compounds.
In
conclusion, the present study demonstrated that ethanol extract has intrinsic
antipyretic and analgesic activity which needs to be investigated with more
information on the bioactive principles responsible for the action. The results
indicate that the ethanol extract of C. filiformis
leaves possesses
significant antipyretic and analgesic activity.
|
Treatment |
Dose |
Rectal
Temperature (ºC) |
||||
|
0.0h |
0.5h |
1.0h |
1.5h |
2.0h |
||
|
Control |
- |
38.15±0.14 |
38.08±0.2 |
38.23±0.15 |
38.48±0.18 |
38.02±0.23 |
|
Paracetamol |
10mg/kg |
38.17±0.21 |
36.8±0.18* |
35.93±0.12* |
35.5±0.18* |
35.45±0.15* |
|
C. filiformis |
25mg/kg |
38.03±0.1 |
37.67±0.12 |
37.1±0.10* |
36.83±0.10* |
37.03±0.10* |
|
50mg/kg |
38.12±0.1 |
37.42±0.10* |
36.78±0.15* |
36.4±0.18* |
36.53±0.20* |
|
|
100mg/kg |
38.28±0.23 |
37.08±0.18* |
36.12±0.15* |
35.78±0.13* |
35.55±0.15* |
|
* Significantly different
from the control at P<0.05,
Table 2 Effects of
ethanol extract of C. filiformis on thermal stimulus induced pain (Hot Plate Test) in mice
|
Treatment |
Dose |
Licked
Time Recorded in Second |
|||
|
0.0h |
0.5h |
1.0h |
1.5h |
||
|
Control |
- |
2.21±0.20 |
2.26±0.20 |
2.39±.20 |
2.19±0.25 |
|
Standard |
10mg/kg |
2.23±0.31 |
5.80±0.80* |
9.40±0.35* |
15.84±0.59* |
|
C. filiformis |
25mg/kg |
2.32±0.20 |
2.89±0.15 |
4.89±0.25* |
5.2±0.34* |
|
50mg/kg |
2.17±0.25 |
3.09±0.18 |
5.08±0.29* |
7.41±0.29* |
|
|
100mg/kg |
2.16±0.27 |
3.61±0.18* |
6.58±0.16* |
10.96±0.47* |
|
* Significantly different
from the control at P<0.05, Standard drug – Diclofenac Sodium
Table 3 Effects of ethanol extract of C. filiformis on pain threshold in hot tail flick test in
mice
|
Treatment |
Dose |
Tail
Flick Response in Second |
|||
|
0.0h |
0.5h |
1.0h |
1.5h |
||
|
Control |
- |
1.93±0.17 |
1.81±0.20 |
1.93±0.20 |
1.90±0.19 |
|
Standard |
10mg/kg |
1.96±0.27 |
5.05±0.28* |
7.54±0.36* |
10.38±0.54* |
|
C. filiformis |
25mg/kg |
2.07±0.16 |
2.56±0.16* |
3.81±0.19* |
4.47±0.20* |
|
50mg/kg |
1.89±0.18 |
2.68±0.22* |
4.82±0.22* |
6.33±0.35* |
|
|
100mg/kg |
1.83±0.20 |
3.33±0.06* |
6.01±0.22* |
8.51±30* |
|
* Significantly different
from the control at P<0.05, Standard drug – Diclofenac Sodium
ACKNOWLEDGEMENTS:
The
authors acknowledge to Director of Oriental College of Pharmacy, Bhopal (M.P.),
India, for providing all the facilities and successfully completion of work.
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Received
on 09.06.2012
Modified on 15.06.2012
Accepted on 25.06.2012
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Research
J. Science and Tech. 4(3): May-June
2012: 129-131