Anti-Inflammatory and Anti-Nociceptive Activity of Adiantum capillus

 

Vikas Gupta1*, Parveen Bansal1, Pawan Kumar2 and Gurpreet Kaur3

1National Institute of Ayurvedic Pharmaceutical Research, Patiala, India2Govt. Polytechnic College for Girls, Patiala, India           3Akal College of Pharmacy and Technical Education, Mastuana Sahib, Sangrur, India

*Corresponding Author E-mail: vikas_4308@rediffmail.com

 

ABSTRACT:

The present study was designed to evaluate both anti-inflammatory and antinociceptive activity of the ethanolic extract of dried leaves of Adiantum capillus (ACEE), using the Carrageenan-induced paw edema method in rats and Tail-flick method in mice, respectively, at various dose levels. The maximum anti-inflammatory effect of the extract was found to be at 300 mg/kg in carrageenan test and this effect was equivalent to phenylbutazone (PBZ) (80 mg/kg, orally) (p<0.05). The extract also demonstrated marked antinociceptive activity at a dose of 300 mg/kg and the effect was comparable to that of standard drug, Ibuprofen (100 mg/kg orally) (p<0.05). The results of this study have established the anti-inflammatory activity and antinociceptive activity of leaf extracts of Adiantum capillus.

 

KEYWORDS: Adiantum capillus, Paw edema method, Adiantaceae, Tail-flick method.

 


 

INTRODUCTION:

Adiantum capillus-veneris L. (Adiantaceae) is a common fern widely distributed throughout the world. The dried whole plant is used as an antipyretic and diuretic, and also in the treatment of bronchitis in folklore medicine in China1. In Northern India, leaves of A.capillus are used for curing the inflammation of throat during cough1. Adiantum species are used for chest complaints, cough, expectorant, increase lactation, colds, aid kidney function, antiparasitic, dandruff, general cure-all. The fresh or dried leafy fronds are antidandruff, antitussive, astringent, demulcent, depurative, emetic, weakly emmenagogue, emollient, weakly expectorant, febrifuge, galactogogue, laxative, pectoral, refrigerant, stimulant, sudorific and tonic2. In Nepal, a paste made from the fronds is applied to the forehead to relieve headaches and to the chest to relieve chest pains. The plant is best used fresh, though it can also be harvested in the summer and dried for later use 3, 4. However, there is no experimental evidence for antinociceptive and anti inflammatory activities with respect to the leaves of this plant. Hence, in the present study, an attempt was made to investigate the anti-inflammatory and antinociceptive effects of the ethanolic extract of dried leaves of Adiantum capillus in experimental animals.

 

MATERIAL AND METHODS:

Plant Material:

The fresh leaves of Adiantum capillus were collected from the surrounding area of Punjab, India and were authenticated by Dr. R.C. Mathad, Professor Department of Dravyaguna, K.L.E.S’s Shri B.M. Kankanawadi Ayurvedic Medical College, Shahapur, Belguam, India.

 

Drugs:

Carrageenan (Sigma-Aldrich, USA), Phenylbutazone and Ibuprofen (MS Pharmaceuticals and Laboratory Ltd., Jalandhar, India) were procured from local market.

 

Preparation of extract:

Dried leaves of Adiantum capillus (500 g) were chopped into small pieces, and soaked overnight in 1.5 litre of 95% ethanol. This suspension was filtered and the residue was re suspended in an equal volume of 95% ethanol for 48 h and filtered again. The two filtrates were pooled and the solvents were evaporated in a rotary evaporator (Perfit, India) at 40-50oC under reduced pressure. A dark semisolid (greenish black) mass was obtained. It was stored below 4oC until further used. When needed, the extract was suspended/ dissolved in desired solvent and used.

 

Animals:

Male Wistar rats and Swiss mice of either sex weighing 150–200g and 20-25g respectively were used in the present study. The animals were kept at room temperature allowing food and water ad libitum and were exposed to normal day and night light cycles. The study was approved by Institutional Animal Ethical Committee.


Table 1: Anti-inflammatory activity of ACEE on carrageenan induced rat paw edema

Groups

Dose (mg/kg)

% inhibition of paw volume

1h

2h

3h

4h

24h

Control

0.0

0.00

0.00

0.00

0.00

0.00

ACEE

100

8.41±0.27

13.71±0.45

17.22±0.31*

23.18±0.23*

10.34±0.22

ACEE

200

12.57±0.64

21.59±0.09*

32.11±0.11*

26.90±0.29*

11.12±0.39

ACEE

300

20.41±0.11*

30.80±0.18*

34.38±0.63*

33.25±0.87*

16.71±0.12

PBZ

80

31.22±0.14*

35.02±0.07*

36.92±0.44*

34.02±0.90*

24.96±0.13*

Values are Mean±SEM (n=6); one way ANOVA. *p<0.05 compared to control, ACEE= Adiantum capillus ethanolic extract, PBZ= Phenylbutazone.

 

 

Table 2: Antinociceptive activity of ACEE on tail-flick assay of mice

Groups

Dose (mg/kg)

% Maximum Possible Effect

1h

2h

3h

Control

0.0

0.00

0.00

0.00

ACEE

100

8.19±0.29

11.22±0.21

13.38±0.54

ACEE

200

12.16±0.26

24.19±0.17 *

31.74±0.10*

ACEE

300

22.01±0.18*

31.19±0.19*

33.59±0.18*

Ibuprofen

100

23.05±0.36*

32.89±0.21*

35.33±0.36*

Values are Mean±SEM (n=6); one way ANOVA. *p<0.05 compared to control, ACEE= Adiantum capillus ethanolic extract.

 

 


Anti-inflammatory activity:

Experimental animals were divided randomly into five groups with six animals in each group (n=6).

Group I- carragenan treated control group receiving normal saline.

Group II- IV - pre-treated with ACEE in doses 100, 200 and 300 mg/kg, orally, respectively.

Group V - pre-treated with standard drug Phenylbutazone 80 mg/kg, orally.

 

Carrageenan-induced rat paw edema:

The ethanolic extract of Adiantum capillus (ACEE) was evaluated for the anti-inflammatory activity5. Acute inflammation was produced by sub-plantar injection of 0.1 ml of 1% carrageenan in normal saline in the right hind paw of the rats, 1h after the administration of the drug/extract. The paw volume was measured by using plethysmometer (Ugo Basile, Italy) at the intervals of 1, 2, 3, 4 and 24 h after the carrageenan injection. Phenylbutazone (80mg/kg, orally.) was used as standard drug. The anti-inflammatory activity was calculated as percentage inhibition of carragenan induced paw edema using the following formula6.

Percent inhibition=100-[(edema volume in treated/edema volume in control) x 100].

 

Antinociceptive activity:

Experimental animals were divided randomly into five groups with six animals in each group (n=6).

Group I- control group receiving normal saline.

Group II- IV - pre-treated with ACEE in doses 100, 200 and 300 mg/kg, orally, respectively.

Group V - pre-treated with standard drug Ibuprofen 100 mg/kg, orally.

 

Tail flick method:

The analgesic activity was determined by radiant heat Tail-flick method in mice7. Ibuprofen (100 mg/kg orally) was used as standard drug. Tail-flick latency was assessed by the analgesiometer (Inco, India). The strength of the current passing through the naked nichrome wire was kept constant at 5A. The distance between heat source and the tail was 1.5 cm and the application site of the heat on the tail was within 2 cm, measured from the root of the tail. Cut-off reaction time was 10s to avoid any tissue injury during the process. Tail-flick latency was measured after 1h of the administration of the drug/extract.

 

Tail flick latencies were converted to maximum possible effect (MPE), according to the following formula:

MPE (%) = 100× (post-extract latency− pre-extract latency)/ (cut-off time− pre-extract latency).

Statistical analysis

 

Results are expressed as mean ± SEM. The statistical significance of the observed data was determined by One Way Analysis of Variance (ANOVA) followed by Dunnet’s test and results were reported to be statistically significant at p < 0.05.

 

RESULTS:

Carrageenan-induced rat paw edema: The anti inflammatory effect of the ethanolic extract of leaves of Adiantum capillus is shown in Table 1. The extract at the oral dose of 300mg/kg showed good results and caused a significant inhibition in the carrageenan induced rat paw edema. The maximum inhibition in edema volume was noted to be 34.38% as comparable to the standard drug, phenylbutazone, which caused maximum inhibition of 36.92% (p<0.05). The extract at lower doses of 100 and 200 mg/kg orally showed lesser anti-inflammatory effect.

 

Tail flick method: The antinociceptive effect of the ethanolic extract of the leaves of Adiantum capillus is shown in Table 2. The extract at the dose of 300mg/kg orally showed 33.59 % of Maximal Possible Effect (%MPE) which was comparable to standard drug, ibuprofen which showed 35.33% of Maximal Possible Effect at dose of 100 mg/kg, orally.(p<0.05)

 

DISCUSSION:

Adiantum capillus is an important medicinal plant which is used in traditional medicine to treat many diseases2. The carrageenan-induced paw edema in rats is believed to be bi-phasic8. The first phase of inflammation is due to the release of histamine and serotonin and the second phase is due to the release of bradykinin, protease, prostaglandins and lysosomes9. Therefore, it can be concluded that inhibitory effect of ACEE on carrageenan-induced paw edema could be due to the inhibition of cyclooxygenase leading to the inhibition of prostaglandin synthesis. In Tail-flick method for antinociceptive activity indicated that ACEE increased the stress tolerance capacity of the animal possibly due to involvement of a higher centre10. However, further work for the isolation of active constituents and elucidating the exact mechanism underlying the observed pharmacological effects is recommended.

 

REFERENCES:

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2.       Grieve M. A Modern Herbal. C.F. Leyel, Penguin, London. 1985.

3.       Chiej R. The MacDonald Encyclopaedia of Medicinal Plants. MacDonald Book Publishers, London and Sydney. 1984; ISBN 0-356-10541-5.

4.       Launert E. Edible and Medicinal Plants. Hamlyn, 1981; ISBN 0-600-37216-2.

5.       Winter CA, Risley EA and Nuss GW. Carrageenan-induced edema in hind paw of the rat as an assay for the anti-inflammatory drugs. Proc. Soc. Exp. Biol. Med. 1962; 111: 544-547.

6.       Chu D and Kovacs BA. Anti-inflammatory activity in Oak gall extracts. Arch. Internat. Pharmacodynamics. 1977; 230: 166–176.

7.       D’Amour FE and Smith DL. A method for determining loss of pain sensation. J. Pharmacol. Exp. Ther. 1941; 72: 74-79.

8.       Vinegar R, Schreiber W and Hugo R. Biphasic development of carrageenan edema in rats. J. Pharmacol. Exp. Ther. 1969; 166: 96-103.

9.       Crunkhorn P and Meacock SC. Mediators of the inflammation induced in the rat paw by carrageenan. British J. Pharmacol. 1971; 42: 392-402.

10.     Whittle BA. The use of changes in capillary permeability in mice to distinguish between narcotic and non-narcotic analgesics. British J. Chemother. 1964; 22: 246-253.

 

 

 

Received on 21.09.2009       Modified on 15.11.2009

Accepted on 19.12.2009      © RJPT All right reserved

Research J. Pharm. and Tech. 3(2): April- June 2010; Page 432-434