Evaluation of Some Biological Activities of <i>Albizia lebbeck</i> Flowers

doi:10.4236/pp.2013.46068

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Pharmacology & Pharmacy, 2013, 4, 473-477

http://dx.doi.org/10.4236/pp.2013.46068 Published Online September 2013 (http://www.scirp.org/journal/pp)

473

Evaluation of Some Biological Activities of Albizia lebbeck

Flowers

Mohamed Farag1*, Ali El Gamal1,2, Ashraf Kalil1,2, Adnan Al-Rehaily1, Omar El Mirghany3,

Kamal El Tahir3

1Pharmacognosy Department, Faculty of Pharmacy, King Saud University, Riyadh, KSA; 2Pharmacognosy Department, Faculty of

Pharmacy, Mansoura University, Mansoura, Egypt; 3Pharmacology Department, Faculty of Pharmacy, King Saud University, Riyadh,

KSA.

Email: *[email protected]

Received May 17th, 2013; revised June 21st, 2013; accepted July 5th, 2013

which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

ABSTRACT

Reviewing the current literature for the importance of the plant Albizia lebbeck L. growing worldwide revealed

many biological interests. However, the species growing in Saudi Arabia has not received due attention. The pre-

sent study was undertaken to study antipyretic, analgesic, estrogenic and anti-inflammatory activities of five different

fractions from successive extraction of Albizia lebbeck flowers: n hexane, dichloromethane, ethyl acetate, n-butanol as

well as the 70% total alcohol. The flowers showed reasonable antipyretic, analgesic, estrogenic and anti-inflammatory

activities.

Keywords: Herbal Medicine; Albizia lebbeck; Anti-Inflammatory Effect; Antipyretic Effect; Analgesic Effect;

Estrogenic Effect

1. Introduction

The genus Albizia (Fabaceae) comprises approximately

150 species, mostly trees and shrubs native to tropical

and subtropical regions of Asia and Africa [1]. Albizia

lebbeck was imported many years ago from India and

well adapted to the harsh environmental conditions of the

central part of Saudi Arabia. The current literature re-

vealed that some plants of the genus Albizia have great

medicinal values. A. lebbeck is a tree well known in the

Indian subcontinent for its range of uses. A. lebbeck is

used in Indian folk medicine to treat several inflamma-

tory pathologies such as asthma, arthritis and burns [2]. A.

lebbeck inhibited the passive cutaneous anaphylaxis and

mast cell degranulation in rat. In addition, it could protect

the sensitized guinea pig from antigen induced anoxic

convulsion [3]. Recently, it was found that the alcoholic

extract of A. lebbeck has antihistaminic property, by neu-

tralizing the histamine directly or due to corticotrophic

action as evidenced by raising cortisol levels in plasma

[4]. Moreover, Saponins of A. lebbeck have been claimed

to be useful in treatment of Alzheimer’s and Parkinson’s

diseases [5]. Leaves have been claimed to have anticon-

vulsant activity [6] and nootropic effect [7] which may

be due to the presence of certain important compounds

like alkaloids and flavanoids. Moreover, the aqueous

extract of A. lebbeck leaves showed antioxidant activity

in diabetic rats [8]. The saponins of the seeds of A. leb-

beck exhibited antiovulatory properties. The seeds had

anti-fertility effect on male rats [9] and antidiarhoeal

activity studied on conventional rodents models of diar-

rhea [10]. In addition, the flowers are being commonly

used to treat anxiety, depression and insomnia in tradi-

tional Chinese medicine [11]. In this study, we report

evaluation of antipyretic, analgesic, estrogenic and anti-

inflammatory activities of different extracts of A. Leb-

beck flowers.

2. Materials and Methods

2.1. Plant Material

The air-dried flowers of A. lebbeck were collected from

Riyadh district, Saudi Arabia in Spring 2008. The plant

was identified and kindly authenticated by Professor Dr.

Ahmad Alfarhan, Department of Botany, College of

Science, King Saud University. A voucher specimen was

deposited at the Pharmacognosy Department, College of

*Corresponding author.

Evaluation of Some Biological Activities of Albizia lebbeck Flowers

474

pharmacy, King Saud University.

2.2. Extraction and Isolation

The air-dried flowers were ground to a coarse powder. A

sample of 700 gm was soaked in 70% ethanol for 3 days

with occasional shaking. This process was repeated four

times until complete exhaustion. The alcoholic extract

was then concentrated to dryness under reduced pressure

at 40˚C using a rotary vacuum evaporator. The crude

dried alcoholic extract (95 gm) was then liquefied in wa-

ter-alcohol mixture (20:80) and successive extracts were

prepared using shaking in separating funnel with n-hex-

ane, dichloromethane, ethyl acetate and n-butanol, re-

spectively.

2.3. Animals

Healthy male adult Swiss albino mice, weighing between

20 - 25 g and albino Wistar rats were obtained from the

Experimental Animal Care Centre, College of Pharmacy,

King Saud University, Riyadh. The conduct of experi-

ments and the procedure of sacrifice (using ether) were

approved by the Ethics Committee of the Experimental

Animal Care Society, College of Pharmacy, King Saud

University, Riyadh, Saudi Arabia.

2.4. Screening for Antipyretic Activity

Six male albino mice, weighing 20 - 25 g, were fasted

overnight before the experiments. Pyrexia was induced

by a subcutaneous injection of 20% w/v brewer’s yeast

suspension (10 ml/kg) into the animal’s dorsum region.

Seventeen hours after the injection, the rectal tempera-

ture of each mouse was measured using a digital ther-

mometer. Only mice that showed an increase in tem-

perature of at least 0.7˚C were used for experiments. A

dose of 1 g/kg of each extract, except n-butanol extract

which was administered in dose of 0.25 g/kg, was ad-

ministered intraperitoneally, and the temperature meas-

ured at 15, 30, 60, 120 and 180 minutes after injection.

Water ad libitum was used as a negative control and as-

pirin (200 mg/kg) was used as a positive control [12].

2.5. Screening for Analgesic Activity—Hot Plate

Method

Male albino white mice, weighing 20 - 25 g, were used.

They were gently placed on a hot plate thermostatically

maintained at 55˚C [13]. The time in sec at which the

animals displayed nociceptive responses exhibited as

licking of the front paws or fanning (blowing) the hind

paws was recorded and the animals were removed from

the plate. A cut-off time of 40 sec was used to avoid

damage of the paws. Crude extracts (500 mg/kg) and a

reference analgesic drug, aspirin (200 mg/kg) were ad-

ministered (i.p) [14].

2.6. Screening for Estrogenic Activity

Two groups each of three immature female albino rats,

weighing 100 g, were housed under a temperature and

light controlled room. They were maintained in a well

ventilated animal quarter. They had free access to water

and commercially available food. To determine estro-

genic activity, a dose of 500 mg of each crude extract/

kg/day was administered intraperitoneally in 1 ml saline

for 3 consecutive days and on the 4th day animals were

sacrificed and uteri were removed and weighed. Normal

saline was used as a negative control and 17-ß-estradiol

(0.32 μg/animal/day) was used as a positive control [15].

2.7. Screening for Anti-Inflammatory

Activity—Carrageenan-Induced

Inflammation

Carrageenan-induced rat hind paw oedema was induced

following the method of [16]. Initially the volumes of the

hind paws of male Wistar rats, weighing 150 - 200 g,

were measurd using hydroplethysmometer (Model 7150,

Ugo Basile, Caemerio, Italy). For this purpose each paw

was marked at the level of the lateral malleoulus and then

dipped gently into the 0.45% NaCl fluid in the chamber

of plethysmometer. This instrument measures the volume

of the paw in ml. Then 0.1 ml of carregeenin (2% w/v in

sterile saline) was injected in one paw under the planter

aponeurosis. The paw volume was then measured hourly

for 4 - 5 hours. Inflammation (or oedema) was expressed

as volume (in ml) increase above the original volume of

the paw or as a percentage increase in the paw volume.

Animals were then injected (i.p) with various doses of

the crude extracts 60 min. before injection of carrageenan.

Then carrageenan was injected as described above and

the paw volume determined hourly for 4 - 5 hours there-

after. Diclofenac sodium was used as standard drug at

concentration of 20 mg/kg of body weight. The influence

of the treatment on the induced inflammation was evalu-

ated [14].

2.8. Statistical Analysis

Values are given as arithmetic means ± standard devia-

tion of the mean (S. D. M.). Data was statistically ana-

lyzed by using the Student’s t-test or ANOVA as appro-

priate. Significance with respect to control group.

3. Results

3.1. Antipyretic Activity

The effects of the different extracts administered at doses

of 1 g/kg except the n-butanol extract which was admin-

istered at a dose 0.25 g/kg (i.p) are shown in Table 1. All

of the treatments decreased the body temperature sig-

nificantly. The maximum decrease of 8˚C was shown by

the dichloromethane extract.

Evaluation of Some Biological Activities of Albizia lebbeck Flowers 475

Table 1. Results of antipyretic study.

A. lebbeck Extract Basal body

temperature

Temperature

after 90 min

Decrease in

temperature

(˚C)

n-butanol (0.25 g/kg) 37.1 34.8 2.3 ± 0.2*

Dichloromethane (1 g/kg) 36.6 28.6 8.0 ± 0.4*

Ethyl acetate (1 g/kg) 36.6 31.6 5 ± 0.9*

Total alcohol (1 g/kg) 36.8 32.1 4.7 ± 0.2*

Aqueous (1 g/kg) 36.6 33.9 2.7 ± 0.9*

n-hexane (1 g/kg) 37.2 35.5 1.7 ± 0.2*

Aspirin (200 mg/kg) 37.2 34.1 3.1 ± 0.7*

The values marked with asterisks differ significantly from the control group,

(*p < 0.05), Number of animals = 5.

3.2. Analgesic Activity

The effect of the different extracts of A. Lebbeck on pain

sensation was tested using hot plate method [13]. Ad-

ministration of the different extracts at doses of 1 g/kg I.p,

except n-butanol extract which was administered in dose

of 0.25 g/kg, induced variable increases in the pain

threshold in the hot plate test. The percentage increases

of pain threshold are shown in Table 2. A reference an-

algesic drug, aspirin (200 mg/kg) was administered as a

positive control [14].

3.3. Estrogenic Activity

Table 3 shows the effects of different extracts of A. leb-

beck on the uteri of immature rats. Normal saline was

used as a negative control and 17-ß-estradiol (0.32 μg/

animal/day) was used as a positive control [15].

3.4. Anti-Inflammatory Activity

Administration of the different A. lebbeck inflorescences

extracts to rats in doses of 1 g/kg I.P (except the n-bu-

tanol extract 0.25 g/kg) with experimental carrageenan-

induced inflammation suppressed inflammation to vari-

ous degrees. The best anti-inflammatory activity was

observed 2 hours after administration of carrageenan. All

extracts were administered 1 hour before carrageenan.

The results are shown in Table 4.

4. Discussion

4.1. Antipyretic Activity

Screening of the antipyretic activity of ethanolic extract

of A. lebbeck seeds was already reported [17], but nothing

was reported concerning flowers growing in Saudi Ara-

bia. By screening of the flowers, it was found that di-

chloromethane and ethyl acetate fractions have signifi-

cant decreases in fever dropped by 8˚C & 5˚C, respec-

tively. Moreover, total alcohol, n-butanol, aqueous and

n-hexane extracts showed less activity than those men-

Table 2. Results of analgesic activity screening.

Reaction time on hot plate

A. lebbeck Extract

Zero time 90 min. after

administration

% increase in

pain threshold

n-butanol (0.25 g/kg) 6.4 ± 0.1 7.4 ± 1.6 14.2 ± 7.6

Dichloromethane (1 g/kg)7 ± 0.2 8.8 ± 0.7 25.7 ± 2.9*

Ethyl acetate (1 g/kg) 7 ± 0.7 8 ± 0.9 14.2 ± 2.2

Total alcohol (1 g/kg) 6.6 ± 0.4 7.6 ± 0.2 15.1 ± 1.1

Aqueous (1 g/kg) 6.8 ± 0.2 6.6 ± 0.7 Not active

n-hexane (1 g/kg) 6.8 ± 0.4 7.6 ± 0.2 8.8 ± 0.9

Aspirin (0.2 g/kg) 5.7 ± 0.2 9.6 ± 0.9 68.4 ± 8

The values marked with asterisks differ significantly from the control group,

(*p < 0.05), Number of animals = 5.

Table 3. Results of estrogenic activity screening.

A. lebbeck Extract

Mean ratio of

uterine horns to

body weight

% change in uterine

weight/total body ratio

Control 0.00547 ± 0.0018 -

n-butanol

(500 mg/kg/day)- Very toxic (Not tested)

Dichloromethane

(500 mg/kg/day)0.00504 7.8% decrease

Ethyl acetate

(200 mg/kg I.P) 0.00409 25.2% ± 5.8% decrease*

Total alcohol

(500 mg/kg/day)0.01144 ± 0.0002 109.141% ± 2.2% increase**

Aqueous

(500 mg/kg/day)0.00574 4.9% increase

n-hexane

(500 mg/kg/day)0.00520 4.93% decrease

The values marked with asterisks differ significantly from the control group,

(*p < 0.05), (**p < 0.001), Number of animals = 4.

Table 4. Results of of anti-inflammatory activity screening.

A. lebbeck Extract

% suppression of inflammation

2 hours after carrageenan

administration

n-butanol (0.25 g/kg) Zero

Dichloromethane (1 g/kg) 71.6 ± 8.5%*

Ethyl acetate (1 g/kg) 60.3 ± 7.3%*

Total alcohol (1 g/kg) 33.9 ± 6.4%*

Aqueous (1 g/kg) 37.7 ± 10.2%*

n-hexane (1 g/kg) 50.9 ± 3.1%*

Diclofenac sodium (20 mg/kg) 67.4 ± 5.4%

The values marked with asterisks differ significantly from the control group,

(*p < 0.05), Number of animals = 3; N. B. Maximum paw edema 2 hours

after carrageenan intra-plantar injection was 0.53 ± 0.05 ml.

tioned above by dropping the temperature by 2.3˚C,

4.7˚C, 2.7˚C and 1.7˚C, respectively as shown in Table 1.

4.2. Analgesic Activity

It was reported that the bark and seeds extracts of A. leb-

Evaluation of Some Biological Activities of Albizia lebbeck Flowers

476

beck have analgesic activity but flowers extracts didn’t

take due attention [18]. For flowers extracts as shown in

the Table 2, a maximum of 25.7% increase in the reac-

tion time was shown by the dichloromethane extract. The

aqueous extract was inactive. Maximum increases in the

pain threshold were observed 90 minutes after admini-

stration of each extract.

4.3. Estrogenic Activity

Estrogens are steroid hormones with important functions

in the regulation of specific sexual processes in the fe-

male. It was reported that methanolic pod extract of A.

lebbeck has antifertility effect but nothing was reported

about estrogenic effect [19]. At a dose of 500 mg/kg I.P.

to immature rats, only total alcohol extract caused sig-

nificant increase in the ratio of weight of the two uterine

horns to the total body weight by 109.14%. This is a very

high percentage and indicates the potent estrogenic ac-

tivity of the total alcohol extract that can point to further

researches. Whereas, the ethyl acetate extract exerted

significant depression of uterine weight/body weight

ratio.

4.4. Anti-Inflammatory Activity

Screening of the anti-inflammatory activity of seeds and

bark of A. lebbeck was reported but nothing about flow-

ers of species growing in Saudi Arabia was reported

[17,18]. The best extract which showed anti-inflamma-

tory activity was the dichloromethane extract with 71.6%

followed by the ethyl acetate extract with 60.3% inhibi-

tors.

In conclusion, these findings clearly suggest that flow-

ers of Albizia lebbeck growing in Saudi Arabia have in-

teresting biological activities and we recommend further

research for isolation of the components responsible for

the estrogenic and anti-inflammatory activities.

5. Acknowledgements

This research project was supported by a grant from the

research Center of the Center for Male Scientific and

Medical Colleges in the King Saud University.

REFERENCES

[1] A. M. Migahid, “Flora of Saudi Arabia: ALBIZIA; Al-

bizia L.,” 3rd Edition, Riyadh University Publication,

Riyadh, 1989, p. 8.

[2] Government of India, The Ministry of Health & Family

Welfare, “Ayurvedic Pharmacopoeia of India,” The Con-

troller of Publication, Delhi, 2001, pp. 201-202.

[3] C. C. Baruach, P. P. Gupta, G. K. Patnaik, D. K. Kul-

shreshtha and B. N. Dhawan, “Anti Anaphylactic and

Mast Cell Stabilizing Activity of Albizzia lebbeck,” In-

dian Journal of Veterinary Medicine, Vol. 21, No. 2,

1997, pp. 127-132.

[4] N. P. Babu, P. Pandikumar and S. Ignacimuthu, “Anti-

Inflammatory Activity of Albizia lebbeck Benth., an Eth-

nomedicinal Plant, in Acute and Chronic Animal Models

of Inflammation,” Journal of Ethnopharmacology, Vol.

125, No. 2, 2009, pp. 356-360.

doi:10.1016/j.jep.2009.02.041

[5] K. Sanjay, “Saponins of Albizia lebbek in Alzheimer’s

and Parkinson’s Disease,” Indian Journal of Natural

Products, Vol. 19, No. 1, 2003, pp. 42-48.

[6] V. S. Kasture, S. B. Kasture and S. C. Pal, “Anticonvul-

sant Activity of Albizzia lebbeck Leaves,” Indian Journal

of Experimental Biology, Vol. 34, No. 1, 1996, pp. 78-80.

[7] S. D. Chintawar, R. S. Somani, V. S. Kasture and S. B.

Kasture, “Nootropic activity of Albizzia lebbeck in Mice,”

Journal of Ethnopharmacology, Vol. 81, No. 3, 2002, pp.

299-305. doi:10.1016/S0378-8741(02)00140-X

[8] C. R. Resmi, M. R. Venukumar and M. S. Latha, “Anti-

oxidant Activity of Albizzia lebbeck (Linn.) Benth. in Al-

loxan Diabetic Rats,” Indian Journal of Physiology and

Pharmacology, Vol. 50, No. 3, 2006, pp. 297-302.

[9] Y. N. Singh, H. Bisht and D. Panday, “Effect of Dry Seed

Extract of a Medicinal Plant Albizzia lebbeck on Testicu-

lar and Epididymal Protein Profiles of Rat,” Himalayan

Journal of Environment and Zoology, Vo. 5, No. 1, 1991,

pp. 94-108.

[10] S. E. Besra, A. Gomes, L. Chaudhary, J. R. Vedasiromoni

and D. K. Ganguly, “Antidiarrhoeal Activity of Seed Ex-

tract of Albizzia lebbeck Studied on Conventional Rodent

Models of Diarrhe,” Phytotherapy Research, Vol. 16, No.

6, 2002, pp. 529-530. doi:10.1002/ptr.961

[11] J. Kang, C. H. Huo, Z. Li and Z. P. Li, “New Ceramides

from the Flower of Albizia julibrissin,” Chinese Chemical

Letters, Vol. 18, No. 2, 2007, pp. 181-184.

doi:10.1016/j.cclet.2006.12.042

[12] Z. Yongna, R. Wantana, B. Pisit, L. Zhongkun and Z.

Rongping, “Analgesic and Antipyretic Activities of the

Aqueous Extract of Urtica macrorrhiza in Experimental

Animals,” Fitoterapia, Vol. 76, No. 1, 2005, pp. 91-95.

doi:10.1016/j.fitote.2004.10.018

[13] G. E. Leighton, M. A. Johnson, K. G. Meecham, R. G.

Hill and J. Hughes, “Pharmacological Profile of PD

117302, a Selective K-Opioid Agonist,” British Journal

of Pharmacology, Vol. 92, No. 4, 1987, pp. 915-922.

doi:10.1111/j.1476-5381.1987.tb11398.x

[14] K. El Tahir, “A Guide to Drug Discovery: Directions for

Pharmacological Screening for New Synthetic and Natural

Compounds Leading to Discovery of New Medicines,” El

Tahir Publication, Riyadh, 2007, pp. 100-105.

[15] A. Kumar and P. Pakrasi, “Estrogenic and Antiestrogenic

Properties of Clomiphene Citrate in Laboratory Mice,”

Journal of Bioscience, Vol. 20, No. 5, 1995, pp. 665-673.

doi:10.1007/BF02703306

[16] G. S. Duncan, S. H. Peers, F. Carey, R. Forder and R. J.

Flower, “Calcium Antagonistic and Antiarrhythmic Ac-

tions of CPU-23, a Substituted Tetrahydroisoquinoline,”

British Journal of Pharmacology, Vol. 109, No. 1, 1993,

pp. 113-119. doi:10.1111/j.1476-5381.1993.tb13539.x

Evaluation of Some Biological Activities of Albizia lebbeck Flowers

477

[17] N. Verma and R. K. Srivastav, “Analgesic, Antipyretic

and Anti-Inflammatory Activities of Albizia lebbeck

Seeds,” Pharmacology Online, Vol. 3, 2011, pp. 1209-

1216.

[18] A. Saha and M. Ahmed, “The Analgesic and Anti-In-

flammatory Activities of the Extract of Albizia lebbeck in

Animal Model,” Pakistanian Journal of Pharmaceutical

Science, Vol. 22, No. 1, 2009, pp.74-77.

[19] R. S. Gupta, J. S. Kachhawa and R. Chaudhary, “Antifer-

tility Effects of Methanolic Pod Extract of Albizzia leb-

beck (L.) Benth in Male Rats,” Asian Journal of Androl-

ogy, Vol. 6, No. 2, 2004, pp. 155-159.