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.

 


Table 1 Antipyretic activity of ethanol extract of C. filiformis on Brewer’s yeast induced pyrexia in rats

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.

 

REFERENCES

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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