Journal of Ethnopharmacology 76 (2001) 1 – 9
www.elsevier.com/locate/jethpharm
Antiulcer activity of Opuntia ficus indica (L.) Mill. (Cactaceae):
ultrastructural study
E.M. Galati a,*, M.T. Monforte a, M.M. Tripodo b, A. d’Aquino a, M.R. Mondello a
b
a
Pharmaco-Biological Department, Vill. SS.Annunziata, 98168, Messina, Italy
Department of Organic and Biological Chemistry, Vill. S.Agata, 98166 Messina, Italy
Received 14 August 2000; received in revised form 2 January 2001; accepted 12 January 2001
Abstract
In Sicily folk medicine, Opuntia ficus indica (L.) Mill. cladodes are used for the treatment of gastric ulcer. We studied the effect
of administration of lyophilized cladodes on experimental ethanol-induced ulcer in rat. In this paper, we report the ultrastructural
observations of gastric mucosa. The ultrastructural changes were observed by trasmission electronic microscopy (TEM)
confirming the protective effect exercised by administration of lyophilized cladodes. Pre-treatment test in rats revealed a protective
action against ethanol-induced ulcer. Probably, the mucilage of Opuntia ficus indica is involved. © 2001 Elsevier Science Ireland
Ltd. All rights reserved.
Keywords: Ethanol-induced ulcer; Folk medicine; Gastric protection; Opuntia ficus indica (L.) Mill.; Transmission electronic microscopy
1. Introduction
Opuntia ficus indica (L.) Mill. is a plant indigenous to
the dry region of the new world whose cultivation has
been suggested for arid land because of its special
adaptative mechanism and capacity to produce
biomass. Moreover O. ficus indica fruits (prickly pears)
are considered of great economic importance in Sicily.
Within the project, supported by Regione Siciliana,
tending to exploit O. ficus indica raw material, we
studied some biological activities (Trovato et al., 2000)
of waste matter (unmarketable fruits and cladodes),
which constitutes a problem for environmental pollution due to fermentation phenomena. In particular, we
assayed diuretic, hypocholesterolemic, anti-inflammatory and cicatrizant activity (Galati et al., 2000; Galati
et al., in press a, Galati et al., in press b, Mondello et
al., 2000).
The cladodes of O. ficus indica are used in folk
medicine in the treatment of gastric mucosal diseases
(Meyer and Mc Laughlin, 1981; Barbera and Inglese,
* Corresponding author.
1993). Therefore, the aim of the present work is to
study the antiulcer activity of lyophilized cladodes of O.
ficus indica on an experimental ethanol-induced ulcer in
rat. In this paper, we also report the results of ultrastructural observations of the gastric mucosa of a rat
treated with lyophilized cladodes.
2. Materials and methods
2.1. Plant material
Opuntia ficus indica lopping cladodes (modified stems
in cacti) were obtained from a cultivation located in
San Cono (CT, Sicily) in May 1999. The identity of the
plant was confirmed by bibliographic data (Tutin et al.,
1968; Pignatti, 1982). Voucher specimens are deposited
at the Pharmaco-Biological Department at the University of Messina.
The fresh tissue, deprived from epidermis and
glochides, was homogenized in a blender and
lyophilized at once. The lyophilized fraction suspended
in water (20 g/100 ml), was given to rats, by gavage, at
a dose of 1 g/kg, in a volume of 1.5 ml/100 g.
0378-8741/01/$ - see front matter © 2001 Elsevier Science Ireland Ltd. All rights reserved.
PII: S 0 3 7 8 - 8 7 4 1 ( 0 1 ) 0 0 1 9 6 - 9
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E.M. Galati et al. / Journal of Ethnopharmacology 76 (2001) 1–9
Table 1
Ulcer scoring scale (Magistretti et al., 1988)
0=no lesion
1=1–3 small lesions (]10 mm length)
2= 1–3 large lesions (]10 mm length)
3 =1–3 thickened lesions
4=more than 3 small lesions
5=more than 3 large lesions
6=more than 3 thickened lesions
Fig. 1. Stomach of a control rat showing hyperemia and lesions
caused by administration of 90% ethanol.
2.2. Animals
Male Wistar rats, weighing 180–200 g, kept in controlled stabulation (temperature 229 2°C; humidity
6094%; natural light), maintained on a standard diet
(S. Morini Mill rat GLP) and water ad libitum, were
used.
2.3. Ulcer induction
The rats were divided into four groups of six animals
each and treated by gavage in the morning.
The first group of rats (control) received orally the
ulcerogenic agent, EtOH 90% (Del Soldato et al., 1984),
at a dose of 0.5 ml/rat.
The second group of rats received O. ficus indica
lyophilized cladodes.
The third group received lyophilized cladodes. The
ulcers were induced by administration of oral ethanol
(0.5 ml) 1 h after lyophilized administration (preventive
treatment).
The fourth group received the ulcerogenic agent
orally (0.5 ml). After 15 min, the rats were administered
O. ficus indica lyophilized cladodes (curative treatment).
One hour later, all the rats were anesthetized using
ether anesthesia, and the stomachs were removed,
opened along the great curvature and delicately washed
with saline solution so as not to remove the mucus
layer from the mucosa surface.
2.4. Macroscopic obser6ations
Fig. 2. Stomach of rat treated with lyophilized cladodes of O. ficus
indica (1 g/kg), before 90% ethanol administration: Preventive treatment.
For the macroscopic observations, the number,
lengths and severity of ulcers were noted and scored on
an arbitrary 0–6 point scale (Magistretti et al., 1988)
(Table 1).
The ulcer index (U.I.) of each stomach was the sum
of its scores.
The ulcer index was reported as arithmetic means9
S.E. The significance of differences between means was
evaluated by Student’s t-test for unpaired data. PB
0.05, versus control, was taken as significant.
2.5. Ultrastructural obser6ations
Fig. 3. Stomach of treated rat with lyophilized cladodes of O. ficus
indica (1 g/kg), after 90% ethanol administration: Curative treatment.
After macroscopic examination of the stomach, fragments were taken along the small curve, not in the
lesion site, in order to study histological modifications.
The fragments were fixed in 2.5% glutaraldehyde and
sodium 0.1 M cacodylate 0.1 M buffer for 2 h at 4°C.
After washing in the same buffer and dehydration by
gradual ethanol (30–100°), the samples were included
in EPON 812.
The ultrathin sections, obtained by the ultramicrotom Ultratom V of the LKB, were stained with uranyl
acetate and lead citrate, according to Reynolds (1963)
and observed using a TEM Philips CM10.
E.M. Galati et al. / Journal of Ethnopharmacology 76 (2001) 1–9
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Fig. 4. Ultrastructural observations of rat gastric mucosa sections observed by transmission electronic microscopy (TEM). O. ficus indica
lyophilized cladodes-treated rats (a, b); ethanol-treated rats (c, d).
3. Results
3.1. Macroscopic obser6ations
In the control rats that received only ethanol, intense
and widespread gastric hyperemia and a thickened lesion were evident (Fig. 1), and the ulcer index (U.I.
8.249 1.5) was calculated.
The pretreatment test, with lyophilized cladodes of
O. ficus indica, revealed a protective action against
ethanol-induced ulcer. The stomach showed an aspect
close to normality. In fact, a significant reduction in
gastric hyperemia in both number and severity of the
lesion was observed (Fig. 2). The ulcer index significantly decreased to 1.489 1.3 (P B 0.05) with respect to
the control (U.I. 8.249 1.5).
O. ficus indica cladodes produced a reduction in the
ulcer index (U.I. 3.259 2.0) (P B 0.05), also when it
was administered as curative, after ethanol administration, but there was no time sufficient to heal the gastric
mucosa (Fig. 3)
3.2. Ultrastructural obser6ations
The ultrastructural changes of gastric mucosa were
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E.M. Galati et al. / Journal of Ethnopharmacology 76 (2001) 1–9
Fig. 4. (Continued)
observed by trasmission electronic microscopy (TEM)
to confirm the antiulcer effect exercised by administration of lyophilized cladodes.
The results revealed some ultrastructural differences
between the groups of rats that were treated respectively with:
1. only O. ficus indica lyophilized cladodes;
2. only ethanol (EtOH 90%);
3. O. ficus indica lyophilized cladodes+ EtOH (preventive treatment);
4. EtOH + O. ficus indica lyophilized cladodes (curative treatment).
E.M. Galati et al. / Journal of Ethnopharmacology 76 (2001) 1–9
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Fig. 5. Ultrastructural observations of rat gastric mucosa sections observed by transmission electronic microscopy (TEM). Preventive O. ficus
indica treatment (a, b, c); curative O. ficus indica treatment (d, e, f).
A. The mucosa of the rats treated only with O. ficus
indica cladodes shows a normal morphology of the
gastric mucosa. Ultrastructurally, it shows characteristic functional aspects of the normal mucosa: the parietal cells highlight a turnover of the canaliculi with an
initial and final stage of secretion; the chief cells appear
to be rich in zymogen granules, with typical fusion of
membrane and endoplasmic reticulum normally represented (Fig. 4a and b).
B. In the rats treated only with the ulcerogenic agent,
we observe numerous alterations in the different cytotypes, with an injury that can be defined as type III,
according to the Lacy and Ito (1982) classification. The
ultrastructural observation shows the degeneration of
the superficial epithelium (Fig. 4c). Few cells seem in
fact very lengthened and full of secretion granules,
others are completely empty, and in a few cases, they
dissolve in the organ lumen. Moreover, it is possible to
observe a wide intra-cellular space that is characteristic
in cases of exfoliation. The most evident morphologic
data are from the parietal cells with dilated secretion
canaliculi and microvilli of various lengths (Fig. 4d);
even the junction complexes are reduced, and on the
cytoplasmic membrane, many introflections are present.
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Fig. 5. (Continued)
In the chief cells, the distribution of secretion granules
seems normal, and the reticulum is well developed, but
we do not observe the typical omega figures present in
the excretion phase.
C. The gastric mucosa of rats treated with lyophilized
cladodes followed by the ulcerogenic agent (EtOH 90%)
shows, in general, an aspect close to normality. The
ultrastructural observation shows a tendency to normalization of different cytotypes, even if the intercellular relationship remains damaged. In the parietal cells,
the nucleus at the center is surrounded by mitochondria
with well-defined crests, and the secretion canaliculi are
E.M. Galati et al. / Journal of Ethnopharmacology 76 (2001) 1–9
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Fig. 5. (Continued)
full of microvilli of varying lengths (Fig. 5a and b). The
chief cells show granules of zymogen partially fused
among themselves and with the membrane, where they
form the omega figure that is characteristic of the
normal cell physiology (Fig. 5c).
D. In the rats that underwent treatment with ulcerogenic agent (EtOH 90%) followed by lyophilized
cladodes, ultrastructural observation of the gastric mucosa reveals degenerating cells in the superficial epithelium (Fig. 5d). In the gland, the chief cells are occupied
by numerous secretion granules in the apex, surrounded
by tubules and cisterns of the rough endoplasmic
reticulum. These formations are even more evident in
the basal portion, where the spherical nucleus is present
(Fig. 5e).
The parietal cells have a central nucleus surrounded
by numerous tightly packed mitochondria with numerous crests, and the canaliculi have numerous microvilli
(Fig. 5f).
The intraglandular tunica is dilated because of the
presence of fibroblasts and collagene fibres. Even the
intercellular space is wide.
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4. Discussion
Acknowledgements
From the results of this work, it is evident that acute
administration of O. ficus indica lyophilized cladodes
generally maintains the cytoarchitecture of the gastric
mucosa in the normal arrangement of its components,
although it is possible to observe an induction to exfoliation of the superficial epithelium near the glandular
apex, which is closed normally.
Ethanol treatment induces a direct damage of gastric
mucosal cells, probably by the development of free
radicals and hyperoxidation of lipid (Puurunen et al.,
1980; Pihan et al., 1987; Ito et al., 1993). Moreover, in
the stomach, ethanol causes solubilization of mucus
constituents and depresses tissue levels of proteins,
leading to flow stasis in gastric blood (Szabo et al.,
1986). The preventive treatment with O. ficus indica
lyophylized cladodes blocks the ulcerative effect of
ethanol.
It is possible to suppose an involvement of O. ficus
indica mucilages in the observed effects. The mucilage
may prevent penetration of the necrotizing agent into
the gastric mucosa. Perhaps, it forms a protective layer
and averts the deep necrotic lesions and the extensive
exfoliation of surface epithelium induced by ethanol.
Probably, the mucilage, a high MW acid polysaccharide
(Trachtenberg and Mayer, 1981) mainly formed by
arabinogalactan and galacturonic acid (Saag et al.,
1975), can act synergetically with defense factors of
gastric mucosa. The polymer present in O. ficus indica
is probably a glycoprotein (Paulsen and Lund, 1979).
This arabinogalactan protein has the potential to interact with macromolecules or small ligand of gastric
mucosa (Clarke et al., 1979). O. ficus indica mucilage is
a negatively charged polyelectrolyte, strongly viscous
because of the negative charges causing strong intermolecular repulsion, resulting in expansion of the
molecules. The viscosity is influenced by pH and Ca++
concentration (Trachtenberg and Mayer, 1982), and in
the gastric lumen, the gelation properties change because of the changes in the conformation of the
molecule. This effect on molecular shape and conformation could be related to protective activity on the
gastric mucosa.
The results are different in preventive and curative
treatment. In the curative treatment, epithelial cells of
gastric mucosa look injured since, after administration
of the ulcerogenous agent, acute treatment with
lyophilized cladodes probably had no time to restore
mucosal defensive factors.
In conclusion, the O. ficus indica lyophylized
cladodes show a significant antiulcer activity in our
experimental model. Our previous study indicates that
lyophylized cladodes have a significant anti-inflammatory activity, too (Galati et al., 2000). New experiments
are under way to correlate the two effects.
Financial support for this research was provided by
Regione Siciliana (Italy).
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