Content uploaded by Vakhtang Mshvildadze
Author content
All content in this area was uploaded by Vakhtang Mshvildadze on Nov 14, 2019
Content may be subject to copyright.
Phytomedicine 12 (2005) 440–444
Acute anti-inflammatory activity of four saponins isolated from ivy:
alpha-hederin, hederasaponin-C, hederacolchiside-E and
hederacolchiside-F in carrageenan-induced rat paw edema
A. Gepdiremen
a,
, V. Mshvildadze
b
,H.Su
¨leyman
a
, R. Elias
c
a
Department of Pharmacology, Medical Faculty, Ondokuz Mayis University, TR-55050 Kurupelit, Samsun-Turkey
b
Institute of Pharmacochemistry, 36, St.P.Sarajishvili, 380059 Tbilisi, Georgia
c
Laboratory of Pharmacognosy and Homeopathy, Pharmacy Faculty of Mediterranean, University of Marseille, 27 Bd.,
J.Moulin, 13385 Marseille, France
Received 28 October 2003; accepted 1 April 2004
Abstract
The anti-inflammatory potential of a-hederin (monodesmoside) and hederasaponin-C from Hedera helix,and
hederacolchisides-E and -F (bidesmosides) from H. colchica was investigated in carrageenan-induced acute paw edema
in rats. Saponins and indomethacin were given orally in concentrations of 0.02 and 20 mg/kg body wt. For the first
phase of acute inflammation, indomethacin was found as the most potent drug. a-hederin and hederasaponin-C were
found ineffective, while hederacolchisides-E and -F showed slight anti-inflammatory effects on the first phase. For the
second phase of acute inflammation, indomethacin and hederacolchiside-F were determined as very potent
compounds. a-hederin was found ineffective for the second phase, either. Despite hederasaponin-C and -E were
found effective in the second phase of inflammation, they were not found as effective as indomethacin and
hederacolchiside-F. As a conclusion, hederasaponin-C, -E and -F, may exert their anti-inflammatory effects by
blocking bradykinin or other inflammation mediators. The latter affect may occur via affecting prostaglandin
pathways. Regarding the structure activity relationship, it is likely that sugars at C
3
position and Rha7-Glc1-6Glc
moiety at C
28
position are essential for the acute anti-inflammatory effect.
r2005 Elsevier GmbH. All rights reserved.
Keywords: Saponins of ivy, Hedera helix; Anti-inflammatory activity; Carrageenan; a-hederin; Hederasaponin; Hederacolchiside
Introduction
Hedera helix (Hedera helix L.) is a well-known plant
as ivy or English ivy, and is a member of Araliaceae
family. Especially the fresh form of leaves and fruits are
toxic, cause gastrointestinal irritation, bloody diarrhea
and death (Baytop, 1984). The best known effect of this
plant is to cause contact dermatitis (Garcia et al., 1995).
Additionally, antibacterial (Cioaca et al., 1978), anti-
helmintic (Julien et al., 1985), leishmanicidic (Majester-
Savornin et al., 1991), in vitro antispasmodic (Trute et
al., 1997), antifungal (Moulin-Traffort et al., 1998)and
acute and chronic anti-inflammatory (Su
¨leyman et al.,
2003) effects of H. helix extracts were reported. On the
ARTICLE IN PRESS
www.elsevier.de/phymed
0944-7113/$ - see front matter r2005 Elsevier GmbH. All rights reserved.
doi:10.1016/j.phymed.2004.04.005
Corresponding author. Tel.: +90 5354883305;
fax: +90 3624576041.
E-mail address: akcahang@omu.edu.tr (A. Gepdiremen).
other hand, H. colchica K. Koch is a less known member
and only antifungal and antiprotozoal activities were
investigated by now (Mshvildadze et al., 2000). Also,
hederacolchiside A1 was tested against proliferation of
human carcinoma and melanoma, recently (Barthomeuf
et al., 2002). It has been demonstrated that bidesmosides
were more active in in vivo experiments than mono-
desmosides (Julien et al., 1985).
It is well known that to investigate the effects of
drugs on the acute phase of inflammation, models
induced by pro-inflammatory agents such as carragee-
nan, dextrane, formaldehyde, serotonin, histamine
and bradykinin in rat paws are employed (Campos
et al., 1995). Carrageenan is perhaps the most com-
monly used and well studied of these phlogistics
(Leme et al., 1973) producing a maximal edema in 3 h.
While the carrageenan model is typically associated
with activation of the cyclo-oxygenase pathway and is
sensitive to glucocorticoids and prostaglandin synthesis
antagonists, the early phase of the carrageenan response
is due to the release of serotonin and histamine (DiRosa
et al., 1971).
In this study, we tested the possible anti-inflammatory
effects of the saponins named a-hederin (AH) and
hederasaponin-C (HsC) extracted from H. helix, and
hederacolchiside-E (HcE) and hederacolchiside-F (HcF)
extracted from H. colchica, in carrageenan-induced
acute paw edema in rats (see Fig. 1).
Material and methods
Plant material
The leaves of H. colchica were collected in the Bagdati
region of West Georgia, in September 2000, and the
leaves of H. helix were collected in Marseille-France, in
September 1999. The materials were identified by Riad
Elias, a staff member of the Laboratory of Pharmacog-
nosy and Homeopathy, Pharmacy Faculty of Mediter-
ranean University, Marseille, France. Vaucher specimen
of the leaves of H. colchica is kept in the Department of
Pharmacobotany, Institute of Pharmacochemistry, Tbi-
lisi, Georgia (leaves no: 70996). Vaucher specimen of the
leaves of H. helix is kept in the Laboratory of
Pharmacognosy, Mediterranean University, Marseille,
France (leaves no: 135797).
Isolation and characterization of hederasaponin C
and a-hederin have been described in previous reports
(Elias et al., 1991). The BuOH extract (2.5 g) of the
leaves of H. helix L., was subjected to low-pressure
chromatography (LPC) on Rp 18, with gradient of
MeOH in H
2
O to yield pure hederasaponin C (1 g).
Alpha-hederin was obtained by alkaline hydrolysis of
hederasaponin C. Hederacolchisides E and F were
isolated from the methanolic extract of leaves of H.
colchica K. Koch, as it was described by other authors
(Dekanosidze et al., 1984). The structures of isolated
ARTICLE IN PRESS
30 29
27
CH2R2
23
R1O
24
25 CO2R3
22
20
12
1
3
17
28
Fig. 1. The chemical structures of triterpene glycosides isolated from H. colchica K. Koch and H. helix L. (Araliaceae).
Name R
1
R
2
R
3
Hederacolchiside E (HCE) Rha 1-2 [Glcl-4]Ara H Rha 1-4 Glcl-6Glc
Hederacolchiside F (HCF) Rha 1-2 [Glcl-4]Ara OH Rha 1-4 Glcl-6Glc
Hederasaponin C (HSC) Rha 1-2 Ara OH Rha 1-4 Glcl-6Glc
Alpha-Hederin (A-H) Rha 1-2 Ara OH H
Ara: a-L-arabinopyranosyl, Rha: a-L-rhamnopyranosyl and Glc: b-D-glucopyranosyl
A. Gepdiremen et al. / Phytomedicine 12 (2005) 440–444 441
compounds were established on the base of MS and
1
H and
13
C NMR methods and the data are in
good agreement with literature values. Optical rotation
for: AH: +9.681(EtOH), HsC: +78.8 (MeOH), HcE:
22.28 (MeOH), HcF: 0 (MeOH). Purity of isolated
compounds were determined by HPLC method: AH
and HsC have 99% purity, while HcE and HcF have
98% of purity. For detailed chemical structure, please
refer to Fig. 1.
Animals
In this study, 42 adult male Wistar albino rats
weighing 180–210 g, and obtained from Atatu
¨rk Uni-
versity, Faculty of Medicine, Department of Pharma-
cology, Experimental Animal Laboratory, were used.
The rats were fed with standard laboratory chow and
tap water before the experiment. The animal laboratory
was equipped with automatic temperature (22711C)
and lighting controls (14 h light/10 h dark). The rats
were divided into groups, each containing 7 individuals.
The investigation conforms with the Guide for the Care
and Use of Laboratory Animals published by the US
National Institutes of Health (NIH Publication No. 85-
23, revised 1996) and the procedures were approved by
the University ethics committee.
Anti-inflammatory studies
Anti-inflammatory effects of AH, HsC, HcE and HcF
were investigated in an aseptic arthritis model, in
0.02 mg/kg (dissolved in 1 ml of water), while indo-
methacin, in 20 mg/kg (dissolved in 1 ml of water) doses
were given to the rats orally by feeding tube as positive
controls. For control animals, 1 ml of water was
administered in the same protocol. The ratio of the
anti-inflammatory effect of the extracts were calculated
by the following equation: Anti-inflammatory activity
(%) ¼(1D/C)100, where Drepresents the percentage
difference in paw volume after the compounds were
administered to the rats, and Crepresents the percen-
tage difference of the volume in the control group.
Carrageenan-induced paw edema
The compounds were given once daily for 2 days. Two
hours after the final administration of the compounds,
0.1 ml (1%, w/v) carrageenan solution in distilled water
was subcutaneously injected into the plantar surface of
the right hind paw. The paw volume was measured with
a plethysmometer three times; once before injection, and
then, 1 and 4 h following the carrageenan administra-
tion. The anti-inflammatory activities in animals that
received AH, HsC, HcE and HcF were compared
with that of indomethacin and the control groups (see
Table 1).
Statistical analysis
Values are presented as mean7SEM. Independent
samples-ttest and analysis of variance (ANOVA,
Dunnett method) were used for the evaluation of data
and po0:05 was accepted as statistically significant.
Results
HsC, HcE and HcF were found to have anti-
inflammatory effects in carrageenan-induced acute
phase of inflammation. In ANOVA test, F-ratio was
2.36 and F-probability was found as 0.053, for first hour
measurements, while F-ratio was 3.57 and F-probability
was 0.0074, for fourth hour measurements. The most
effective drug was found to be indomethacin
(22.0374.1%, po0:001 in respect to control group)
for first hour measurements. HcE (32.3374.9%,
po0:05 in respect to control group) and HcF
(32.175.4%, po0:05 in respect to control group) were
found slightly effective, and AH and HsC were found
statistically ineffective, in that period. Values represent
ARTICLE IN PRESS
Table 1. Effects of saponins on carrageenan-induced rat paw edema
Treatment Dose (mg/kg) nEdema rate percentage (mean7SEM)
1h p4h p
Control — 7 49.274.5 — 57.475.7 —
Indomethacin 20 7 22.0374.1 o0.001 29.3272.6 o0.001
Alpha hederin 0.02 7 39.674.3 40.05 42.876.7 40.05
Hederasaponin-C 0.02 7 39.275.6 40.05 36.2974.7 o0.01
Hederacolchiside-E 0.02 7 32.3374.9 o0.05 36.9175.1 o0.05
Hederacolchiside-F 0.02 7 32.175.4 o0.05 29.175.2 o0.001
Groups compared to controls, by independent samples-ttest.
A. Gepdiremen et al. / Phytomedicine 12 (2005) 440–444442
percent increase in respect to first measurements of each
group.
For fourth hour measurements, except AH, all the
groups tested has been found effective. Despite the AH
administered group results (42.876.7%) were found
better than the control (57.475.7%), it was statistically
insignificant. Hederacolchiside-F (29.175.2%, po0:001
in respect to control group) was found as effective as
indomethacin (29.3272.6%, po0:001 in respect to
control group), while HsC (36.2974.7%, po0:01 in
respect to control group) and HcE (36.9175.1%,
po0:05 in respect to control group) exerted lesser anti-
inflammatory effects within the fourth hour of inflam-
mation. Values represent percent increase in respect to
first measurements of each group (Table 1).
Discussion
Several triterpene saponins have been tested in several
applications by now. Buddlejasaponin and saikosapo-
nin’s in vivo anti-inflammatory effect on mouse ear
edema (Bermejo-Benito et al., 1998) and zanhasaponin’s
acute and chronic anti-inflammatory effects (Cuellar et
al., 1997) were shown before. It was reported that the
anti-inflammatory effects of several agents result in the
partial inhibition of inflammation mediator release
(Amadio et al., 1993). Subcutaneous injection of
carrageenan into the rat paw produces plasma extra-
vasation (Szolcsanyi et al., 1998) and inflammation,
characterized by increased tissue water and plasma
protein exudation with neutrophil extravasation and
metabolism of arachidonic acid by both cyclooxygenase
and lipoxygenase enzyme pathways (Gamache et al.,
1986).
In the present study, all the saponin groups and
indomethacin, except AH, exerted anti-inflammatory
effects, in respect to control. Despite HcF and indo-
methacin found as effective as each other, regarding the
doses of 20 mg/kg for indomethacin and 0.02mg/kg for
HcF, it may be speculated that HcF is a more potent
compound than indomethacin for the second phase of
inflammation. Contrary the results of AH and HsC
application, HcE and HcF, in very low doses, were
found quite effective to prevent acute phase of
inflammation for the same period. For the first phase
of acute inflammation, we found indomethacin as the
most potent compound, tested in the present study.
There are biphasic effects in carrageenan-induced
edema. The first phase begins immediatelly after
injection and diminishes within 1 h. The second phase
begins at 1 h and remains through 3 h (Garcia-Pastor et
al., 1999). It is suggested that the early hyperemia of
carrageenan-induced edema results from the release of
histamine and serotonin (Kulkarni et al., 1986). The
delayed phase of carrageenan-induced edema results
mainly from the potentiating effects of prostaglandins
on mediator release, especially of bradykinin. Hydro-
cortisone and some NSAIDs strongly inhibit the second
phase of carrageenan-induced edema, but some others
are effective against both phases (Kulkarni et al., 1986).
According to that, indomethacin seems to block both
phases, that means blocking histamin and serotonin
release within the first hour, and it also prevents
releasing some of the inflammatory mediators, via
blocking the prostaglandin’s action within the second
phase. On the other hand, saponin derivatives (except
AH), seem to affect mainly the second phase of acute
inflammation. Despite the fact that both, hederacolchi-
side-E and -C, found slightly effective on that period,
this effect was not as obvious as indomethacin or
hederacolchiside-F. So, it is very likely that, HsC and
HcE exert their anti-inflammatory effects by blocking
bradykinin or other inflammation mediators. The latter
effect of saponins may occur via affecting prostaglandin
pathways. HcF may affect some additional pathways,
similar to indomethacin.
Regarding their chemical structures (Fig. 1), HcE and
HcF with an additional glucose molecule in the sugar
chain at R1 position, are more active than AH and HsC.
So it may be speculated that, the glucose moiety in the
sugar residue connected to the C
3
position of the genin is
crucial for acute anti-inflammatory effect, especially for
the late phase of it. AH, monodesmoside with free
carboxyl group at C-17 position of the aglycone was
determined as a unique ineffective compound in the
present study. It could be supposed that, Rha1-4Glc1-
6Glc residue at R3 position may be responsible of the
anti-inflammatory effect.
The present study shows that HsC and HcE are slight
and HcF is a potent inhibitor of acute inflammation,
especially on the second phase. The mechanism of the
effect may depend on the inhibition of the formation of
some inflammatory mediators. In conclusion, detailed
studies are needed to clarify the mechanism(s) of anti-
inflammatory effects of saponins.
References
Amadio Jr., P., Cummings, D.M., Amadio, P., 1993.
Nonsteroidal anti-inflammatory drugs. Tailoring therapy
to achieve results and avoid toxicity. Postgrad. Med. 93,
73–76.
Barthomeuf, C., Debiton, E., Mshvildadze, V., Kemertelidze,
E., Balansard, G., 2002. In vitro activity of hederacolchisid
A1 compared with other saponins from Hedera colchica
against proliferation of human carcinoma and melanoma
cells. Planta Med. 68, 672–675.
Baytop, T., 1984. Therapy with medicinal plants in Turkey
(past and present). Publications of the Istanbul University,
Istanbul, p. 225.
ARTICLE IN PRESS
A. Gepdiremen et al. / Phytomedicine 12 (2005) 440–444 443
Bermejo-Benito, P., Abad-Martinez, M.J., Silvan-Sen, A.M.,
Sanz-Gomez, A., Fernandez-Matellano, L., Sanchez-Con-
treras, S., Diaz-Lanza, A.M., 1998. In vivo and in vitro
anti-inflammatory activity of saikosaponins. Life Sci. 63,
1147–1156.
Campos, M.M., Mata, L.V., Calixto, J.B., 1995. Expression of
B1 kinin receptors mediating paw edema and formalin-
induced nociception. Modulation by glucocorticoids. Can.
J. Physiol. Pharmacol. 73, 812–819.
Cioaca, C., Margineanu, C., Cucu, V., 1978. The saponins of
Hedera helix with antibacterial activity. Pharmazie 33,
609–610.
Cuellar, M.J., Giner, R.M., Recio, M.C., Just, M.J., Manez,
S., Cerda, M., Hostettmann, K., Rios, J.L., 1997.
Zanhasaponins A and B, antiphospholipase A2 saponins
from an anti-inflammatory extract of Zanha africana root
bark. J. Nat. Prod. 60, 1158–1160.
Dekanosidze, G.E., Djikia, O.D., Vulgalter, M.M., Kemerte-
lidze, E.P., 1984. Triterpene glycosides of Hedera colchica:
structure of hederacolchisides E and F. Khim. Prir. Soedin.
6, 747–749.
DiRosa, M., Giroud, J.P., Willoughby, D.A., 1971. Studies of
the mediators of the acute inflammatory response induced
in rats in different sites by carrageenan and turpentine.
J. Pathol. 104, 15–29.
Elias, R., Diaz-Lanza, A.M., Vidal-Ollivier, E., Balansard, G.,
Faure, R., Babadjamian, A., 1991. Triterpene saponins
from leaves of Hedera helix. J. Nat. Prod. 54, 98–103.
Garcia, M., Fernandez, E., Navarro, J.A., del Pozo, M.D.,
Fernandez de Correz, L., 1995. Allergic contact
dermatitis from Hedera helix L. Contact Dermatitis 33,
133–134.
Garcia-Pastor, P., Randazzo, A., Gomez-Paloma, L.,
Alcaraz, M.J., Paya, M., 1999. Effects of petrosaspongio-
lide M, a novel phospholipase A2 inhibitor, on acute and
chronic inflammation. J. Pharmacol. Exp. Ther. 289,
166–172.
Gamache, D.A., Povlishock, J.T., Ellis, E.F., 1986. Carragee-
nan-induced brain inflammation. Characterization of the
model. J. Neurosurg. 65, 679–685.
Julien, J., Gasquet, M., Maillard, C., Balansard, G., Timon-
David, P., 1985. Extracts of the ivy plant, Hedera helix, and
their anthelminthic activity on liver flukes. Planta Med. 17,
205–208.
Kulkarni, S.K., Mehta, A.K., Kunchandy, J., 1986. Anti-
inflammatory actions of clonidine, guanfacine and B-HT 920
against various inflammagen-induced acute paw oedema in
rats. Arch. Int. Pharmacodyn. Ther. 279, 324–334.
Leme, J.G., Hamamura, L., Leite, M.P., Silva, M.R., 1973.
Pharmacological analysis of the acute inflammatory process
induced in the rats paw by local injection of carrageenan
and by heating. Br. J. Pharmacol. 48, 88–96.
Majester-Savornin, B., Elias, R., Diaz-Lanza, A.M., Balan-
sard, G., Gasquet, M., Delmas, F., 1991. Saponins of the
ivy plant, Hedera helix, and their leishmanicidic activity.
Planta Med. 57, 260–262.
Moulin-Traffort, J., Favel, A., Elias, R., Regli, P., 1998. Study
of the action of alpha-hederin on the ultrastructure of
Candida albicans. Mycoses 41, 411–416.
Mshvildadze, V., Favel, A., Delmas, F., Elias, R., Faure, R.,
Kemertelidze, E., Balansard, G., 2000. Antifungal and
antiprotozoal activities of saponins from Hedera colchica.
Pharmazie 55, 325–326.
Su
¨leyman, H., Mshvildadze, V., Gepdiremen, A., Elias, R.,
2003. Acute and chronic anti-inflammatory profile of the
ivy plant, Hedera helix, in rats. Phytomedicine 10, 370–374.
Szolcsanyi, J., Helyes, Z., Oroszi, G., Nemeth, J., Pinter, E.,
1998. Release of somatostatin and its role in the mediation
of the anti-inflammatory effect induced by antidromic
stimulation of sensory fibres of rat sciatic nerve. Br.
J. Pharmacol. 123, 936–942.
Trute, A., Gross, J., Mutschler, E., Nahrstedt, A., 1997. In
vitro antispasmodic compounds of the dry extract obtained
from Hedera helix. Planta Med. 63, 125–129.
ARTICLE IN PRESS
A. Gepdiremen et al. / Phytomedicine 12 (2005) 440–444444
