8/14/2019 Atta 1998- Plantas Antiinflamatorias
1/8
Journal of Ethnopharmacology 60 (1998) 117124
Anti-nociceptive and anti-inflammatory effects of someJordanian medicinal plant extracts
A.H. Atta a, A. Alkofahi b,*
a Department of Veterinary Basic Sciences, Faculty of Veterinary Medicine, Jordan Uniersity of Science and Technology,
P.O. Box 3030, Irbid, Jordanb Department of Medicinal Chemistry and Pharmacognosy, Faculty of Pharmacy, Jordan Uniersity of Science and Technology,
P.O Box 3030, Irbid, Jordan
Received 15 October 1996; received in revised form 14 July 1997; accepted 4 November 1997
Abstract
The anti-nociceptive effect of ethanolic extract of 11 traditionally used Jordanian plants was studied by using the
acetic acid-induced writhing and hot-plate test in mice. The anti-inflammatory effect of these plants was determined
by xylene-induced ear oedema in mice and cotton pellet granuloma test in rats. Mentha piperita, Cinnamomum
zeylanicum, Apium graeolens,Eucalyptus camaldulentis, andRuta graeolens possess an anti-nociceptive effect against
both acetic acid-induced writhing and hot plate-induced thermal stimulation. M. piperita, Jasminum officinale,
Commiphora molmol, andBeta ulgarispossess an anti-inflammatory effect against acute (xylene-induced ear oedema)
and chronic (cotton-pellet granuloma) inflammation. The anti-nociceptive and anti-inflammatory effects were dosedependent. These data affirm the traditional use of some of these plants for painful and inflammatory conditions.
1998 Elsevier Science Ireland Ltd. All rights reserved.
Keywords: Anti-nociceptive activity; Anti-inflammatory activity
1. Introduction
There are more than 49 plant families having
more than 120 plant species used in Jordanian
traditional medicine, especially among people who
have little or no access to medical assistance(Karim and Quraan, 1986; Al-Khalil, 1995). The
anti-inflammatory and/or the analgesic effects of
Apium graeolens, Beta ulgaris, Eucalyptus
camaldulentis, Jasminum officinale, Mentha piper-
ita, Ruta graeolens, Lactuca satia, Cinnamomum
zeylanicum, and Commiphora molmol have been
described in folk medicine (Al-Jarwani and Khal-ifeh, 1936; Jabour, 1983). Moreover, A. graeo-
lens, M. piperita, and C. molmol have been
traditionally used as anti-spasmodics, anti-* Corresponding author.
0378-8741/98/$19.00 1998 Elsevier Science Ireland Ltd. All rights reserved.
PII S 0 3 7 8 - 8 7 4 1 ( 9 7 ) 0 0 1 3 7 - 2
8/14/2019 Atta 1998- Plantas Antiinflamatorias
2/8
A.H. Atta, A . Alkofahi/Journal of Ethnopharmacology 60 (1998) 117124118
Table 1
List of plants used in the screening for anti-nociceptive and anti-inflammatory activity
Part used Voucher specimen number Yield g/kgScientific name
Dried juice 89 860A. era L. (Liliaceae)
3462A. graeolens L. (Umbelliferae) Seed
9655B. ulgaris L. (Chenopodiaceae) Root
Bark 11C. zeylanicum L. (Lauraceae) 50
114 168Oleo-gum resinC. molmol Engl. (Burseraceae)
Leaf 46E. camaldulentis Dehn. (Myrtaceae) 118
132 80J. officinale L. (Oleaceae) Flower
Seed 58L. satia L. (Compositeae) 104
Leaf 33M. piperita L. (Labiatae) 112
52 38O. syriaca (L.) Rafin (Labiatae) Leaf
Leaf 94R. graeolens L. (Rutaceae) 130
rheumatics, and to relieve teeth and ear pains,
skin diseases and headache (Kotb, 1985). In addi-
tion, these plants have been used for gastrointesti-
nal, respiratory, and reproductive disorders.Moreover, some of them are used as germicides
and vermifuges. The aim of this work is to eluci-
date the anti-nociceptive and anti-inflammatory
effects of 11 Jordanian plants (Table 1) of known
traditional use in relieving pain and inflammation.
2. Materials and methods
2.1. Plant materials
Extracts were prepared from 11 plants com-
monly used in Jordan as analgesics and/or anti-
inflammatory. Most of these plants grow locally.
Samples were obtained either as dried plants from
herbal stores or collected from the wild. The
taxonomic identity of the plants was confirmed by
Professor A. El-Oqlah, Department of Biological
Sciences, Yarmouk University, Irbid, Jordan. A
voucher specimen of each species studied has been
deposited at the Department of Medicinal Chem-
istry and Pharmacognosy, Faculty of Pharmacy,
Jordan University of Science and Technology,
Irbid, Jordan.
2.2. Preparation of plant extract
Each dehydrated plant was ground to a fine
texture and 100 g of the dried plant were repeat-
edly extracted with 80% ethanol. The ethanol
extracts were concentrated under vaccum and
weighed and the residue was used in the experi-ments. The dried plant extracts were freshly dis-
solved or suspended in distilled water just before
administration.
2.3. Acetic acid writhing in mice
The writhing test was performed as described
by Koster et al. (1959). Groups of ten Swiss mice
(five males and five females) were fasted over
night prior to dosing, but had free access to
water. After 30 min of receiving an oral dose of
the extract, each mouse was given intraperitonealy0.7% aquous solution of acetic acid (10 ml/kg b.
wt.) and then placed in an observation box. The
number of writhes were counted for 20 min after
acetic acid injection. The number of writhes in
each treated group was compared to that of a
control saline-treated group.
2.4. Hot-plate test in mice
The method described by Eddy and Leimback
(1953) was applied. Groups of ten mice each were
used. Mice were placed in a 2-l glass beakerplaced on a hot plate maintained at 55C. Latency
to exhibit the nociceptive response such as licking
paws or jumping was determined before and 15,
8/14/2019 Atta 1998- Plantas Antiinflamatorias
3/8
A.H. Atta, A . Alkofahi/Journal of Ethnopharmacology 60 (1998) 117124 119
Table 2
Effect of ethanolic extracts on acetic acid-induced writhes in mice
200 mg/kg 400 mg/kgTreatment groups
Mean+S.D. Protection (%) Mean+S.D. Protection (%)
40.2+10.049.6+12.2Normal saline (10)
38 22.4+3.1***M. piperita (10) 30.2+9.5** 44
21 35.8+
6.0O. syriaca (10) 39.4+
5.3 11
18.8+3.8***48 5325.0+4.3***C. zeylanicum (8)
21 27.5+6.0**E. camaldulentis (8)a 39.2+10* 32
23.0+6.6*** 43A. graeolens (10) 29.6+10.2*** 40
48 20.6+4.6***J. officinale (9) 26.0+5.2*** 49
0 40.4+9.2 063.6+9.4B. ulgaris (10)
24 30.3+4.2**C. molmol (10) 37.5+11.5* 25
24 28.4+6.1**A. era (10) 37.6+10.3* 29
42.0+8.40 049.4+7.5L. satia (10)
32 27.0+4.1***R. graeolens (10)a 3333.8+7.0**
( ) Number of animals.a A dose of 100 and 200 mg/Kg b. wt.
*P0.05, **P0.01, ***P0.001 compared to control.
30, 45, 60, and 75 min after intraperitoneal ad-
ministration of the extract. A cut-off time of 60 s
was selected to avoid tissue damage.
2.5. Xylene-induced ear oedema in mice
Male Swiss mice were divided into groups of
ten mice each. After 30 min of the i.p. injection of
the extract, xylene (0.03 ml) was applied to the
anterior and posterior surfaces of the right ear.
Mice were sacrificed 2 h after xylene application
and both ears were removed. Circular sections of
both treated and untreated ears were taken using
a 7 mm diameter cork borer and weighed. The
difference in weight between left untreated ear
sections and right treated ear section was calcu-
lated (Tang et al., 1984).
2.6. Cotton pellet granuloma in rats
Sterilized cotton pellets of 20 mg weight each,
were impregnated with 0.4 ml of ampicillin
aquous solution. Pellets were implanted subcuta-neously in the groin region of Wistar rats, one on
each side. The drug was given orally by stomach
tube once daily for 6 days starting with the day of
implantation. On day 7, the rats were killed and
the pellets were removed, dried at 70C for 24 h.
The weight of granuloma and adrenal gland was
recorded and compared to that of saline treated
control (Schiatti et al., 1986). All plant extracts
were given in doses of 200 and 400 mg/kg body
weight except E. camaldulentis and R. graeolens
which were given in doses of 100 and 200 mg/kg
based on pilot experiments and appearance of
some toxic manifestations after larger doses.
2.7. Statistical analysis
The data were presented as meanS.D. Sig-
nificance between control and treated groups was
tested by ANOVA.
3. Results
Ethanolic extract ofC. zeylanicum, J. officinale,M. piperita, A. graeolens, and R. graeolens sig-
nificantly (P0.01 and P0.001) reduced thenumber of acetic acid-induced writhes in mice
with a protection percent ranging from 32 to 48
and from 33 to 53 after administration of 200 and
8/14/2019 Atta 1998- Plantas Antiinflamatorias
4/8
A.H. Atta, A . Alkofahi/Journal of Ethnopharmacology 60 (1998) 117124120
Table 3
Effect of ethanolic extracts on time to response of mice to heat stimulation in the hot-plate test
Treatment Doses mg/kg Time to response (s)
groups
45 min 60 min0 min 15 min 30 min 75 min
Normal saline
7.48+
2.4 7.34+
1.9 8.42+
3.8(20) 8.34+
3.25.10 +
1.1 5.66+
1.5
M. piperita
7.72+1.1 8.42+0.4(6) 200 7.9+2.6 4.7+0.2 5.76+1.4 8.28+0.5
10.63+ 3.7** 15.52+6.1***(10) 400 6.1+2.3 7.15+2.1 7.10 +0.5 8.04+2.3
O. syriaca
8.1+0.4 8.3+0.6 7.86+0.4(7) 200 8.2+0.56.2+1.8 6.4+0.8
13.2 +6.24**10.88+1.1*(10) 5.93+0.9400 8.96+2.2*5.6+1.5 5.19+1.4
C. zeylanicum
8.7+0.4 7.0+0.9(5) 200 6.56+0.7 6.0+1.4 8.82+0.9 7.58+0.4
11.55+3.0*** 10.59+0.9*(10) 10.7+1.9*400 9.82+3.6**5.22+1.6 7.9+1.7***
E. camaldulen-
tis
6.4+
1.86.16+
1.5 6.9+
1.8(5) 6.1+
1.08100 7.0+
2.8 5.8+
1.211.51+ 4.4*** 13.4+5.3**(10) 200 5.18+0.5 14.3+3.8*** 17.4+6.9*** 12.8+5.7*
A. graeolens
9.5+1.98.0+2.5 10.1+2.9(5) 7.9+1.8200 5.01+1.0 6.34+1.5**
12.1+3.1**11.2+2.1*(10) 400 5.87+2.0 9.58+4.3*** 9.4+2.2* 10.6+2.3***
J. officinale
6.62+0.9 7.7+0.6(5) 200 7.02+2.5 4.92+1.5 6.9+2.9 6.98+1.2
7.8+1.9 8.08 +2.01(10) 400 4.81+2.4 6.72+2.1 6.28 +3.3 7.88 +3.9
B. ulgaris
46+ 1.2 6.12+0.6 6.9+1.3 7.42+0.9(5) 200 6.3+2.7 4.84+1.4 7
8.81+4.47.27+3.9(10) 7.71+3.2400 6.9+3.14.09+1.8 6.02 +1.3
C. molmol
6.9+0.9 8.16+0.6(5) 200 5.64+0.9 6.44+3.4 7.9+3.0 8.68+0.2
8.01+
2.66.91+
2.66.01+
0.7(10) 400 6.15+
1.56.0+
1.8 5.13+
1.3
A. era
7.5+0.87.6+0.67.3+0.5(5) 6.6+1.2200 6.4+0.6 5.6+0.9
6.64+2.1 6.33+2.6(10) 400 5.03+1.6 6.18+1.4 8.24+3.9 8.8+2.2
L. satia
6.78+2.47.3+1.0 5.8+2.7(5) 6.2+0.3200 6.38+0.4 6.44+0.6
7.7+2.66.89+1.4(10) 400 4.52+1.2 6.7+1.9 7.4+2.0 8.85+1.8
R. graeolens
6.99+2.6 8.9+1.36.1+2.5(5) 6.2+2.6100 6.95+2.3 7.9+2.7*
8.89+0.8* 7.68+1.4 6.49+0.6(10) 8.08+2.4200 4.07+1.8 7.38+2.3**
( ) Number of animals.
*P0.05, **P0.01, ***P0.001 compared to control. Mean+S.D.
400 mg/kg, respectively (Table 2). Ethanolic ex-
tract of E. camaldulentis, C. molmol, and Aloe
era also decreased the number of acetic acid-in-
duced writhes but to a less extent (P0.05 and
P0.01 for the low and high dose, respectively).
The ethanolic extract of O. syriaca, B. ulgaris,
8/14/2019 Atta 1998- Plantas Antiinflamatorias
5/8
A.H. Atta, A . Alkofahi/Journal of Ethnopharmacology 60 (1998) 117124 121
Table 4
Effect of ethanolic extracts on xylene-induced ear swelling in mice
200 mg/kg 400 mg/kg
Weight (mg) Inhibition (%) Weight (mg) Inhibition (%)
3.4+1.44.3+1.9Normal saline
1.7+0.8* 50M. piperita 2.2+1.2* 49
47 2.1+
0.8O. syriaca 2.3+
1.6 38
3.3+1.10 34.9+1.2C. zeylanicum
3.0+1.0 12E. camaldulentisa 3.9+2.7 9
63.2+1.0163.6+0.9A. graeolens
0 1.6+0.3*J. officinale 5.0+6 4.2 53
0 1.7+0.8*B. ulgaris 5.2+6.2 50
501.7+0.5*352.8 +2.1C. molmol
5.0+0.7 0 1.0+0.5** 71A. era
0 2.8+0.8L. satia 5.3+0.23 18
3.4+2.00 05.7+3.0R. graeolensa
a A dose of 100 and 200 mg/kg b. wt.
*P0.05, **P0.01. MeanS.D., n=5.
andL.satia in an oral dose of 200 or 400 mg/kg,showed no significant effect on the number of
acetic acid-induced writhes.
Ethanoilic extract ofC. zeylanicum, E. camald-
ulentis, and A. graeolens produced a highly sig-
nificant (P0.001) increase in the latency to
response of mice to hot plate thermal stimulation.
This effect starts 15 min and persisted for at least
75 min after administration of the plant extract
(Table 3). Mild or no effect was observed by the
small dose. Mild anti-nociceptive effect has also
been observed after administration ofM. piperita,
O. syriaca, and R. graeolens. However, theireffect was either temporary as in R. graeolensor
delayed in onset (45 min) as in M. piperita and O.
syriaca. On the other hand the ethanolic extracts
of J. officinale, B. ulgaris, C. molmol, A. era,
and L. satia showed no anti-nociceptive effect.
Ethanolic extract of A. era in a dose of 400
mg/kg significantly (P0.01) reduced the weight
of xylene-induced ear oedema in mice with a
calculated inhibition of 71%, while the smaller
dose produced no significant effect (Table 4). M.
piperite (both doses), J. officinale, B. ulgaris, and
C.molmolalso significantly (P0.05) reduced thesize of ear oedema with an inhibition percent
ranging from 50 to 53 when used at the high dose
only. O. syriaca, C. zeylanicum, E. camaldulentis,
A.graeolens, L. satia, andR. graeolensshowedno significant effect on xylene-induced oedema in
mice ears.
Ethanolic extract of A. graeolens, B. ulgaris,O.syriaca,C. zeylanicum, J. officinale,C. molmol,
and L. satia in a dose of 400 mg/kg and R.graeolens in a dose of 200 mg/kg significantly
reduced the weight of cotton granuloma in rats
(Table 5). Non of these extracts except A. graeo-lens (P0.05) was effective by the smaller dose.
The most effective extracts was that ofA. graeo-lens and B. ulgaris (P0.001). E. camaldulentis
and A. era have no effect at any of the useddoses. None of the tested plant extracts in the
used doses affect the weight of adrenal glands in
rats.
4. Discussion and conclusion
In the present study, two animal models for
investigation of the anti-nociceptive and anti-infl-
ammatory effects of the selected plants were used.
The hot plate thermal stimulation and the acetic
acid induced writhes in mice were selected toinvestigate the central and peripheral anti-noci-
ceptive effects. Xylene-induced ear swelling in
mice and the cotton pellet granuloma in rats were
8/14/2019 Atta 1998- Plantas Antiinflamatorias
6/8
A.H. Atta, A . Alkofahi/Journal of Ethnopharmacology 60 (1998) 117124122
Table 5
Effect of ethanolic extracts on the weight of granuloma and adrenal glands in rats
Doses (mg/kg) Weight of granuloma (mg)Treatment Weight of adrenal (mg)
Normal control
0.107+0.030 0.046+0.030(8)
0.015+0.011(5) 0.059+0.017
M. piperita0.086+0.017 0.010+0.003(5) 200
400 0.045+0.009**(6) 0.066+0.009
O. syriaca
200 0.076+0.038 0.006+0.004(6)
0.036+0.002(6) 0.054+0.006**400
C. zeylanicum
200 0.057+0.012(6) 0.013+0.007
(6) 0.037+0.0050.075+0.007*400
E. camaldulentis
0.009+0.0040.056+0.021(5) 100
0.060+0.001 0.032+0.020(5) 200
A. graeolens
200 0.037+
0.012*(5) 0.020+
0.0160.046+0.0130.061+0.001***(9) 400
Jasminum officinale
0.017+0.0060.076+0.017(5) 200
400 0.069+0.014*(6) 0.087+0.010
Beta ulgaris
0.009+0.0030.054+0.001(6) 200
400 0.059+0.011***(8) 0.027+0.006
Commiphora molmol
0.060+0.011 0.012+0.003(5) 200
0.071+0.016* 0.024+0.008(5) 400
Aloe era
200 0.071+0.032(5) 0.010+0.003
400 0.083+0.005 0.015+0.001(5)
L. satia
200 0.073+0.024(5) 0.001+0.003
400 0.060+0.014*(5) 0.037+0.002
R. graeolens
0.016+0.0010.080+0.034(6) 100
200 0.047+0.001*(5) 0.029+0.004
( )Number of animals.
*P0.05, **P0.01, ***P0.001. MeanS.D.
selected to represent models of acute (exudative
phase) and chronic (the poliferative phase) inflam-mation respectively.
The present results show that M. piperita, C.
zeylanicum, E. camaldulentis, A. graeolens, and
R. graeolens induced a dose-dependent analgesic
protective effect against both thermal stimuli andthe writhing syndrome indicating central and pe-
ripheral effects. The increased latency to response
of mice to thermal stimuli only after administra-
8/14/2019 Atta 1998- Plantas Antiinflamatorias
7/8
A.H. Atta, A . Alkofahi/Journal of Ethnopharmacology 60 (1998) 117124 123
tion of the ethanolic extract of O. syriaca indi-
cates central analgesic effect only. On the other
hand, J. officinale, A. era, and C. molmol in-
duced an anti-nociceptive effect against acetic
acid-induced writhing only indicating the lack of
central anti-nociceptive effect of these three ex-
tracts.
The present results demonstrate that ethanolicextracts of M. piperita (dose dependent), J.officinalis, B. ulgaris, and C. molmol(large dose
only) possess a dose-dependent anti-inflamma-
tory effect against both acute (exudative) and
chronic (proliferative) inflammation. The activity
of ethanolic extract of A. era against acute infl-
ammation (xylene-induced ear oedema) only in-
dicates possible anti-phlogestic but not
anti-proliferative effect. O. syriaca, C. zey-lanicum, A. graeolens, L. satia, and R. graeo-lens showed an anti-inflammatory effect only
against chronic inflammation induced by cottonpellet granuloma indicating anti-proliferative ef-
fect. E. camaldulentis ethanolic extract in the
used dosage and route of administration showed
no anti-inflammatory effect.
All the tested extracts, except E. camaldulentis,
have variable degrees of both anti-nociceptive
and anti-inflammatory effect. Such an association
is well-known for various non-stroidal autin-infl-
ammatory (NSAI) compounds especially salicy-
late by-products (Reuse, 1978; Beuoist and
Misse, 1979; Famaey, 1983; Gyires et al., 1985).M. piperita, C. zeylanicum, A. graeolens, E.camaldulentis, and R. graeolens have both anti-
nociceptive and anti-inflammatory effects. These
five plant extracts contain common active princi-
ples; volatile oils, resins, and flavenoids (Kotb,
1985). Volatile oils, resins and flavenoids isolated
from other plant extracts have been proved to
posses analgesic and/or anti-inflammatory effect
(Duke, 1992). Therefore, it could be suggested
that the anti-nociceptive and anti-inflammatory
effects of the tested plant extracts may be due to
their content of volatile oils, flavenoids and
resins. From these results, it can be affirmed that
the traditional indication of some of these plantextracts for inflammation and pain associated
with sprains, bruises, wounds, spasmodic colics,
and rheumatic arthritis (Al-Khalil, 1995; Bajpai
and Sant, 1995; Sudarsanam et al., 1995). Fur-
ther investigations are necessary to elucidate the
exact mechanism of the anti-nociceptive and
anti-inflammatory activity of these extracts.
Acknowledgements
The authors would like to thank Jihan Abu-
Zaher for her technical assistance and the Dean-
ship of the Scientific Research in the University
of Science and Technology for financial support
(Grant No. 44/95).
References
Al-Jarwani, M., Khalifeh, O., 1936. Tathkirat Daoud Al-An-
taki. Iben Shakroun library, Egypt (1858), pp. 126, 128,
137, 171.
Al-Khalil, S., 1995. A survey of plants used in Jordanian
traditional medicine. International Journal of Pharmacog-
nosy 33 (4), 317323.
Bajpai, A.J.K., Sant, H.R., 1995. Medicobotany of the
Varansi district, Utter Pardesh, India. International Jour-
nal of Pharmacognosy 33 (2), 172176.
Beuoist, J.M., Misse, J.L., 1979. In: Giroud, J.P., Mathe, G.,
Meyniel, G. (Eds.), Pharmacologic clinique: bases de la
therapeutique 2. Expansion Scientifique Francaise, Paris,
10491091.
Duke, J.A., 1992. Handbook of Biologically Active Phyto-
chemicals and their Activities. CRC Press, Boca Raton,
FL.
Eddy, N.B., Leimback, D., 1953. Synthetic analgesics: II.Dithyienylbutenylamines and dithyienylbutylamines. Jour-
nal of Pharmacology and Experimental Therapeutics 3,
544547.
Famaey, J.P., 1983. In: Blotman, F., Crastes de Paulet, A.,
Simon, L. (Eds.), Biochimie de Inflmmation: Acide Arachi-
donique et ses Derives. Masson, Paris, 174188.
Gyires, K., Furts, S., Miklya, I., Budavari, I., Knoll, J., 1985.
Drugs Under Experimental and Clinical Research 2 (8),
493500.
Jabour, J., 1983. Al-Kanoon in Medicine of Avicenna. Al-
Maaref Library. Beirut, Lebanon.
Karim, F.M., Quraan, S.A., 1986. Medicinal Plants of Jordan.
Yarmouk University, Irbid, Jordan.
Koster, R., Anderson, M., De Beer, E.J., 1959. Acetic acid for
analgesic screening. Federation Proceedings 18, 412.Kotb, F.T., 1985. Medicinal Plants in Lybia. Arab Encyclope-
dia House, Beirut, Lebanon.
Reuse, J., 1978. In: Fabre, J. (Ed.), Therapeutique Medicale,
Flammarion Medicine-Sciences, Paris, pp. 207 219.
8/14/2019 Atta 1998- Plantas Antiinflamatorias
8/8
A.H. Atta, A . Alkofahi/Journal of Ethnopharmacology 60 (1998) 117124124
Schiatti, P., Selva, D., Galliani, G., Baldol, E., Diena, A.,
Glasser, A., Leali, M., Toja, E., 1986. Highly selective
anti-inflammatory and analgesic activity of 3-(1-methyl-
ethyl)-2-(4-methoxyphenyl)-3H-maphth(1,2-d) imidazole, a
new man acidic molecule. Arzneimittele-Forschung 36,
102109.
Sudarsanam, G., Reddy, M.B., Nagaraju, N., 1995. Veterinary
crude drugs in Rayalaseema, Andhra, Pradesh, India. In-
ternational Journal of Pharmacognosy 33 (1), 5260.
Tang, Xican, Lin, Zhigong, Cai, Wen, Chen, Nian, Shen, Li,
1984. Anti-inflammatory effect of 3-acetylaconitine. Acta
Pharmacologica Sinica 5, 8589.
.