Journal of Ethnopharmacology 103 (2006) 66–70

Antisecretory activity of plants used to treat gastrointestinal

disorders in Mexico夽
Claudia Velázquez a,d,∗ , Fernando Calzada a,∗ , Javier Torres b ,
Felipe González c , Guillermo Ceballos d
aUnidad de Investigación Médica en Farmacologı́a de Productos Naturales, Hospital de Pediatrı́a, 2◦ Piso, Centro Médico Nacional Siglo XXI,
IMSS, Av. Cuauhtemoc 330, Col. Doctores, CP 06725, México D.F., Mexico
b Unidad de Investigación Médica en Enfermedades Infecciosas y Parasitarias, Hospital de Pediatrı́a, 2◦ Piso, Centro Médico Nacional Siglo XXI,
IMSS, Av. Cuauhtemoc 330, Col. Doctores, CP 06725, México D.F., Mexico
c Unidad de Investigación Médica en Infectologı́a e Inmunologı́a, Hospital de Infectologı́a, 2◦ Piso, Centro Médico Nacional La Raza,

IMSS, Av. Jacarandas, S/N, CP 02990, México D.F., Mexico

d Instituto Politécnico Nacional, Escuela Superior de Medicina, Plan de San Luis y Dı́az Mirón, CP 11340, México D.F., Mexico

Received 9 August 2004; received in revised form 24 June 2005; accepted 29 June 2005
Available online 19 September 2005

Abstract

Aqueous and methanolic extracts from 26 medicinal plants used in Mexico to treat gastrointestinal disorders were screened to evaluate their
antisecretory activity on cholera toxin-induced intestinal secretion in rat jejunal loops model. Extracts were tested at a dose of 300 mg/kg.
From 56 samples tested, both extracts from Chiranthodendron pentadactylon, Hippocratea excelsa and Ocimum basilicum were the most
potent with inhibition values ranging from 68.0 to 87.6%. On the other hand, the methanolic extract of Geranium mexicanum (aerial parts)
and the aqueous extract of Bocconia frutescens showed the highest activity with inhibition values of 93.4 and 86.0%, respectively. The results
obtained in this study give some scientific support to the use of the Mexican medicinal plants employed for the treatment of gastrointestinal
disorders such as diarrhea.
© 2005 Elsevier Ireland Ltd. All rights reserved.

Keywords: Mexican medicinal plants; Aqueous extracts; Methanolic extracts; Antisecretory activity; Cholera toxin

1. Introduction secretion of transepithelial electrolytes, thus increasing the

osmotic flux of water and ions to the intestinal lumen. Specif-
Diarrheal disease is one of the most common causes ically, heat-labile (LT) and heat-stable (ST) enterotoxins
of morbidity and mortality in many developing countries from Escherichia coli, Vibrio cholerae and Campylobacter
(Guerrant, 1985; Amstrong and Cohen, 1999). It is often jejuni increase net fluid secretion by affecting the enzymes
caused by enterotoxins which are produced by bacteria adenylyl cyclase or guanylyl cyclase in the intestinal mucose
such as enterotoxigenic Escherichia coli, Salmonella typhi, (Guerrant, 1985; Torres et al., 1993; Mutschler et al., 1995;
Salmonella typhimurium, Clostridium difficile, Clostrid- Hör et al., 1995; Torregosa et al., 1996; Raufman, 1998;
ium freundii, Aeromonas hydrophila, Yersinia enterocolitica, Amstrong and Cohen, 1999).
Campylobacter jejuni and Vibrio cholerae. Enterotoxins have To control diarrhea, the treatment of choice is oral rehy-
their effect on the enterocyte functions by stimulating the dration solution (ORS). It has reduced the levels of mortality
in children and elderly by dehydration, but not morbidity for
夽 Part of this work was taken from the Ph.D. thesis of Claudia Velázquez. diarrhea (Amstrong and Cohen, 1999; Turvill et al., 2000).
∗ Corresponding authors. Tel.: +525 627 6900x22410; fax: +525 761 0952. To treat the secretory diarrhea there are some drugs, such as
E-mail address: [email protected] (F. Calzada). racecadotril and loperamide, that decrease intestinal hyper-

0378-8741/$ – see front matter © 2005 Elsevier Ireland Ltd. All rights reserved.
doi:10.1016/j.jep.2005.06.046
 C. Velázquez et al. / Journal of Ethnopharmacology 103 (2006) 66–70 67

secretion. Both drugs have side effects; racecadotril cause pended in 1 mL of sterile water. For the study, aliquots of the
brochospasm, fever and vomiting, and loperamide should not toxin solution were dissolved in a 1× PBS (NaCl 8 g, KCl
be administrated to children younger than 6 years of age and 0.2 g, Na2 HPO4 ·7H2 O 0.115 g, KH2 PO4 0.2 g/L) solution
patients with constipation, and intestinal obstruction, and in with 1% bovine serum albumin (Sigma) to obtain a concen-
acute dysentery caused by bacterial infections because it can tration of 3 ␮g/mL.
mask the symptoms and cause intestinal perforation (Brown,
1979; Rogé et al., 1993; Salazar et al., 2000). 2.4. Antisecretory assay
On the other hand, there are some compounds that showed
inhibitory properties on the intestinal secretion, such berber- The antisecretory activity of the extracts was tested using
ine, chlorpromazine, clonidine, nicotinic acid, indometacin, a method previously described (Torres et al., 1993). Male
somatostatin and ethycrynic acid, but they were not devel- Sprague–Dawley rats (200–250 g) were obtained from the
oped as antidiarrheal drugs (Rabbani et al., 1982; Sack and animal house of the IMSS. The experimental protocols were
Froehlich, 1982; Fedorak and Field, 1987). approved by the Animal Care and Use Committee of Hospital
In Mexico, the use of medicinal plants to treat gas- de Pediatria del Centro Medico Nacional Siglo XXI, IMSS, in
trointestinal disorders such as diarrhea and dysentery is accordance with the guidelines for care and use of laboratory
widespread (Campos, 1991; Aguilar et al., 1994). However, animals. The effect of the extracts was studied on intestinal
most of these plant species have not been investigated from secretion indirectly by measuring the fluid accumulation in
a pharmacological point of view to demonstrate their antise- the intestine following cholera toxin administration to rats.
cretory properties, which could lead to support their use as Two jejunal loops were prepared in the rats and inoculated
antidiarrheal and antidysenteric drugs in traditional medicine. with 3 ␮g/mL of cholera toxin dissolved in 1× PBS with 1%
In this work, we screened 56 aqueous and methanolic bovine albumin. Rats (n = 4 per group by duplicated) were
extracts from 26 Mexican medicinal plants to assess their treated orally with each extract (300 mg/kg in 1 mL of a 2%
antisecretory activity using the cholera toxin-induced intesti- DMSO solution in water), or vehicle (2% DMSO solution
nal secretion in rat jejunal loops model. It is important to in water). Loperamide (10 mg/kg) was used as antidiarrheal
notice that none of these species or their isolated metabolites drug, After 4 h, the animals were sacrificed using ethyl ether.
have been previously evaluated as antisecretory agents. The antisecretory activity of the extracts was measured as
the fluid secretion in the loops and expressed in percentage
of inhibition.
2. Materials and methods
2.5. Statistical analysis
2.1. Plant materials
Values are expressed as mean ± S.E.M. Statistical signif-
The assessed plants were collected by the authors (Calzada
icance was determined using Mann–Whitney U-test. Values
and Velázquez) from different regions in Mexico: Mexico
with p < 0.05 were considered significant.
City, States of Hidalgo, Mexico, Sinaloa, Guanajuato and
Yucatan. They were selected according to their use in Mex-
ican traditional medicine to treat gastrointestinal disorders.
3. Results and discussion
Voucher herbarium specimens (Table 1) have been authenti-
cated by MS Abigail Aguilar, Herbarium IMSSM of Instituto
We tested 56 aqueous and methanolic crude extracts
Mexicano del Seguro Social. Plant species, part used, voucher
obtained from medicinal plants used in Mexican traditional
specimens and the percentage of inhibition of the intestinal
medicine for the treatment of gastrointestinal disorders.
secretion are shown in Table 1.
These were tested at oral doses of 300 mg/kg because at this
2.2. Preparation of crude extracts dose most of the extracts showed antisecretory activity. In
traditional medicine since infusions or decoctions are usually
The air-dried plant material (20 g) was extracted by mac- taken three times per day when diarrhea occurs, our results
eration with 300 mL of MeOH for 1 week. Then the macer- can be related with their traditional use because the used dose
ate was filtered and concentrated under reduced pressure at is approximately one cup of plant tea which is recommended
40 ◦ C. For aqueous extracts, 20 g of air-dried plant material by Mexican people to treat gastrointestinal disorders (Aguilar
was extracted by decoction with 100 mL of distilled water for et al., 1994).
30 min, the solution was filtered and lyophilized. The yields The antisecretory activity was tested using the cholera
are shown in Table 1. toxin-induced intestinal secretion in rat jejunal loops model.
The antisecretory activity of the extracts tested is shown in
2.3. Cholera toxin Table 1. We found that both extracts from Chiranthodendron
pentadactylon, Hippocratea excelsa and Ocimum basilicum
Lyophilized powder (1 mg) of cholera toxin (Sigma) con- were the most active with inhibition values ranging from 68.0
taining approximately 220,000 units/mg of protein was sus- to 87.6% at 300 mg/kg. Methanolic extract of Geranium mex-
68 C. Velázquez et al. / Journal of Ethnopharmacology 103 (2006) 66–70

Table 1
Antisecretory activity of methanolic and aqueous extracts of selected Mexican medicinal plants at 300 mg/kg on intestinal secretion response to cholera toxin
Family Plant specie Part used Voucher number Extract % (w/w) yield % Inhibition
Liliaceae Allium sativum L. EB 14396 Methanolic 1.2 18.4 ± 10.8
Aqueous 10.7 14.9 ± 5.5
Verbenaceae Aloysia triphylla (L’Hér) Britton AP 126110 Methanolic 7.1 7.8 ± 4.7
Aqueous 17.4 80.4 ± 22.8
Annonaceae Annona cherimola Miller S 14401 Methanolic 8.7 79.5 ± 14.6
Aqueous 3.4 50.2 ± 12.9
Asteraceae Artemisa absinthium L. AP 12609 Methanolic 10.8 9.1 ± 2.3
Aqueous 20.7 20.7 ± 11.1
Asteraceae Artemisa ludoviciana Nutt AP 14409 Methanolic 12.7 5.7 ± 2.9
Aqueous 15.9 15.0 ± 6.8
Papaveraceae Bocconia frutescens L. AP 12618 Methanolic 9.7 24.1 ± 15.4
Aqueous 14.3 86.0 ± 9.8
Leguminosae Caesalpinia pulcherrima (L.) Sw. AP 13591 Methanolic 23.0 16.3 ± 6.6
Aqueous 23.8 12.8 ± 3.8
Caricaceae Carica papaya L. S 14397 Methanolic 6.7 20.9 ± 5.9
Aqueous 23.3 13.6 ± 11.7
Chenopodiaceae Chenopodium ambrosioides L., green variety AP 14402 Methanolic 10.7 43.4 ± 6.5
Aqueous 13.8 48.7 ± 11.6
Chenopodiaceae Chenopodium ambrosioides L., red variety AP 14395 Methanolic 13.9 19.7 ± 7.5
Aqueous 21.1 16.7 ± 6.7
Chenopodiaceae Chenopodium murale L. AP 13592 Methanolic 8.3 31.9 ± 5
Aqueous 15.9 32.8 ± 9.9
Sterculiaceae Chiranthodendron pentadactylon Larreat F 14104 Methanolic 17.7 87.6 ± 15.3
Aqueous 12.6 84.8 ± 17.4
Asteraceae Chrysactinia mexicana A. Gray AP 14407 Methanolic 8.4 0
Aqueous 23.1 0
Palmae Cocos nucifera L. EF 14398 Methanolic 29.1 23.4 ± 11.9
Aqueous 19.4 12.5 ± 5.4
Convolvulaceae Dichondra argentea Humb & Bonpl AP 0103m Methanolic 12.1 22.3 ± 13.2
Aqueous 16.5 36.7 ± 12.1
Moraceae Dorstenia contrajerva L. AP 14406 Methanolic 8.6 24.4 ± 16.4
Aqueous 0.3 44.8 ± 5.9
Geraniaceae Geranium mexicanum H. B. & K. AP 14405 Methanolic 15.5 93.4 ± 6.7
Aqueous 31.0 42.1 ± 15.2
Geraniaceae Geranium mexicanum H. B. & K. R 14405 Methanolic 14.6 0
Aqueous 15.6 0
Hippocrateaceae Hippocratea excelsa H. B. & K. R 14394 Methanolic 7.2 80.3 ± 21.3
Aqueous 15.5 75.0 ± 24.9
Verbenaceae Lippia alba (Mill.) N.E.Br. AP 14146 Methanolic 8.5 32.0 ± 9.8
Aqueous 20.4 26.0 ± 9.1
Schizaeaceae Lygodium venustum Sw. AP 13270 Methanolic 11.2 51.6 ± 15.6
Aqueous 12.3 0
Compositae Matricaria recutita L. AP 14399 Methanolic 19.9 25.4 ± 7.6
Aqueous 30.5 24.3 ± 3.6
Labiatae Ocimum basilicum L. AP 14393 Methanolic 10.5 68.7 ± 19.7
Aqueous 23.1 68.0 ± 20.8
Punicaceae Punica granatum L. EF 14403 Methanolic 48.2 55.9 ± 3.6
Aqueous 28.5 19.1 ± 6.9
 C. Velázquez et al. / Journal of Ethnopharmacology 103 (2006) 66–70 69

Table 1 (Continued )
Family Plant specie Part used Voucher number Extract % (w/w) yield % Inhibition
Rutaceae Ruta chalepensis L. AP 14400 Methanolic 23.0 73.7 ± 7.01
Aqueous 23.9 23.6 ± 9.27
Anacardiaceae Schinus molle L. AP 14408 Methanolic 18.4 0
Aqueous 12.8 79.5 ± 17.7
Leguminosae Senna villosa Mills L 14410 Methanolic 9.4 11.9 ± 3.4
Aqueous 35.7 29.3 ± 17.9
Labiatae Thymus vulgaris L. AP 13594 Methanolic 9.4 0
Aqueous 12.0 18.9 ± 10.3
Loperamide – – – – – 43.3 ± 13.1
The antisecretory activity was expressed as the percentage inhibition (mean ± S.D., n = 8) compared to the blank. Loperamide was used as a positive control
(10 mg/kg). AP, aerial parts; EB, bulb epidermis; EF, fruit exocarp; F, flowers; L, leaves; R, roots; S, seed. p < 0.05 is considered to be significant.

icanum (aerial parts) and the aqueous extract of Bocconia frutescens alkaloids have been isolated and the methano-
frutescens were the most active with inhibition values of 93.4 lic and chlorophorm extracts showed antibacterial activity
and 86.0%, respectively. against Vibrio cholera, a causal agent of diarrhea (Perez,
On the other hand, the methanolic extract of Chenopodium 2000; Caballero et al., 2002, 2003). The antiprotozoal or
ambrosioides green variety (aerial parts), Lygodium venus- antibacterial activity seen in the most active extracts of this
tum, Punica granatum and Ruta chalepensis, the aqueous study together with the antisecretory effect are evidence that
extracts of Aloysia triphylla, Chenopodium ambrosioides support the use of Chiranthodendron pentadactylon, Gera-
green variety (aerial parts), Dorstenia contrajerva and Schi- nium mexicanum and Bocconia frutescens in Mexican tra-
nus molle showed inhibitory activity with values rang- ditional medicine to treat diarrhea. It is important to notice
ing from 43.4 to 79.5%. The 87% of the extracts tested that no extract or isolated compound of these species has
showed inhibitory activity of the intestinal secretion; only been previously evaluated to demonstrate their antisecretory
seven extracts did not show activity. In general, among the activity.
researched extracts, the methanolic extracts exhibited the Some studies have been performed in order to find anti-
highest antisecretory activity. secretory compounds from plants used in the traditional
medicine to treat various kinds of diarrheas, in this sense
the extracts of Croton urucurana, Croton lechleri, Berberis
4. Conclusion aristata and Guazuma ulmifolia were studied against the
intestinal secretion caused by Vibrio cholerae toxin. In the
Some of the medicinal plants tested showed interest- cases of Berberis aristata, Guazuma ulmifolia and Cro-
ing antidiarrheal activity in vivo model. Both extracts of ton lechleri their antisecretory compounds were berberine
Annona cherimola, Chiranthodendron pentadactylon, Hip- and oligomeric proantocyanidins, respectively. From Cro-
pocratea excelsa, Ocimum basilicum, Geranium mexicanum ton urucurana saponins, steroids, alkaloids, antocianidins
green variety (aerial parts), methanolic extracts from Ruta and catechins have been isolated. In Guazuma ulmifolia
chalepensis, Lygodium venustum, Punica granatum, aqueous (Hör et al., 1995) and Croton urucurana, proanthocyani-
extracts of Bocconia frutescens, Aloysia triphylla, Dorstenia dins and catechins can probably be associated with their
contrajerva and Schinus molle, showed better antisecretory antisecretory activity (Dutta and Panse, 1962; Sack and
activity than loperamide. These results allows to propose Froehlich, 1982; Fedorak and Field, 1987; Hör et al., 1995;
these species as a potential sources of antisecretory com- Gabriel et al., 1999; Gurgel et al., 2001; Fischer et al.,
pounds and should be therefore subjected to further bioassay- 2004).
guided phytochemicals studies to obtain their active princi- It is important to notice that our active extracts showed
ples. antisecretory activity to 300 mg/kg whereas that in the case of
From the most active species, Chiranthodendron Croton urucurana it showed antisecretory activity at an oral
pentadactylon, several compounds including flavonoids, dose of 600 mg/kg (Gurgel et al., 2001). The active extracts
steroids, hydrocarbons, sugars and gallic acid, which showed found in this work will be an option to develop novel phyto-
moderate activity against the protozoa Giardia lamblia, have drugs useful to treat fluid loss in diarrhea.
been isolated (Lara and Márquez, 1996; Alanis et al., 2003). A The antisecretory compounds isolated from medicinal
bioassay-guided study on the antiprotozoal activity of Gera- plants combined with ORS might be useful in decreasing
nium mexicanum against Entamoeba histolytica and Giardia the mortality caused by dehydration. The results obtained in
lamblia led to the isolation of a steroid, an alkaloid, a sugar this study give some scientific support to the use of selected
and two flavan-3-ols, and (−) epicatechin was found to be medicinal plants in Mexican tradition for the treatment of
the active compound (Calzada et al., 2005). From Bocconia gastrointestinal disorders such as diarrhea.
70 C. Velázquez et al. / Journal of Ethnopharmacology 103 (2006) 66–70

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