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Acta Tropica journal homepage: www.elsevier.com/locate/actatropica
Anti-Trypanosoma cruzi activity of 10 medicinal plants used in northeast Mexico
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Zinnia Judith Molina-Garza a , Aldo Fabio Bazaldúa-Rodríguez b , Ramiro Quintanilla-Licea b , Lucio Galaviz-Silva a,∗ a Universidad Autónoma de Nuevo León, UANL, Facultad de Ciencias Biológicas, Laboratorio de Patología Molecular, Ave. Universidad S/N, Cd. Universitaria, 66451 San Nicolás de los Garza, NL, Mexico b Universidad Autónoma de Nuevo León, UANL, Facultad de Ciencias Biológicas, Laboratorio de Fitoquímica, Ave. Universidad S/N, Cd. Universitaria, 66451 San Nicolás de los Garza, NL, Mexico
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Article history: Received 20 November 2013 Received in revised form 3 April 2014 Accepted 5 April 2014 Available online xxx
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Keywords: Trypanocidal activity Chagas disease Medicinal plants
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1. Introduction
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The aim of this study was to screen the trypanocidal activity of plants used in traditional Mexican medicine for the treatment of various diseases related to parasitic infections. Cultured Trypanosoma cruzi epimastigotes were incubated for 96 h with different concentrations of methanolic extracts obtained from Artemisia mexicana, Castela texana, Cymbopogon citratus, Eryngium heterophyllum, Haematoxylum brasiletto, Lippia graveolens, Marrubium vulgare, Persea americana, Ruta chalepensis and Schinus molle. The inhibitory concentration (IC50 ) was determined for each extract via a colorimetric method. Among the evaluated species, the methanolic extracts of E. heterophyllum, H. brasiletto, M. vulgare and S. molle exhibited the highest trypanocidal activity, showing percentages of growth inhibition between 88 and 100% at a concentration of 150 g/ml. These medicinal plants may represent a valuable source of new bioactive compounds for the therapeutic treatment of trypanosomiasis. © 2014 Published by Elsevier B.V.
Chagas disease is caused by the protozoan parasite Trypanosoma cruzi, and over 100 years after its discovery by Carlos Chagas (Chagas, 1909), this disease continues to represent a major health issue in Latin America. Initially, this disease primarily occurred in rural areas, where the causative agent was transmitted from blood-sucking insects of the Reduviidae family to humans (Cardenas-Sánchez et al., 2003); however, currently, the accidental oral transmission of T. cruzi is becoming increasingly common (Bastos et al., 2010), whereas transmission by vectors and by blood transfusion has substantially decreased throughout Latin America (WHO, 2014). In addition, migration has brought infected individuals to urbanized areas of Latin America and to Europe, Japan, Australia (Schmunis, 2007) and the United States (Carod-Artal et al., 2005; Reisenman et al., 2010; Bern et al., 2011), where infections through non-vectorial routes can occur via blood
∗ Corresponding author. Tel.: +52 81 83524425/+52 83 524425. E-mail addresses: [email protected], [email protected] (L. Galaviz-Silva).
transfusion (Kirchhoff et al., 2006; Galaviz-Silva et al., 2009), organ transplantation, and congenital transmission (Gürtler et al., 2003; Schmunis, 2007). This disease is also known as American trypanosomiasis, and approximately 7–8 million people are currently infected (WHO, 2014). Furthermore, the estimated number of people infected worldwide has declined to 8 million, with an annual incidence rate of 56,000 cases and an estimated 12,000 deaths occurring every year (PAHO, 2013). Mexico is a country with high climatic variety and great biodiversity; this environment provides excellent habitats for the geographic distribution of Triatominae species, which can be found in most Mexican states (Cruz-Reyes and Pickering-Lopez, 2006). In the state of Nuevo León in northeast Mexico, the estimated population at risk of T. cruzi infection is approximately 90,277 individuals (Carabarin-Lima et al., 2013). Only Triatoma gerstaeckeri (Stål) has been characterized as a domiciliary and peridomestic vector, whereas T. neotomae (Neiva), T. lecticularia (Stål) and T. protracta (Uhler) are involved in sylvatic cycles (Martínez-Ibarra et al., 1992; Molina-Garza et al., 2007). Nifurtimox and benznidazole have been used for over 40 years to treat Chagas disease; however, these drugs are only effective during the acute phase of infection, and both pharmaceuticals induce
http://dx.doi.org/10.1016/j.actatropica.2014.04.006 0001-706X/© 2014 Published by Elsevier B.V.
Please cite this article in press as: Molina-Garza, Z.J., et al., Anti-Trypanosoma cruzi activity of 10 medicinal plants used in northeast Mexico. Acta Trop. (2014), http://dx.doi.org/10.1016/j.actatropica.2014.04.006
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significant side effects after long periods of medication usage. Furthermore, certain protozoan strains have developed resistance to treatment with these drugs (Sülsen et al., 2006; Rojas et al., 2010). The development of new, safer and more effective trypanocidal compounds remains a challenge because these drugs are not given high priority by the R&D-based pharmaceutical industry (Troullier et al., 2002; Sülsen et al., 2006). Phytotherapy represents the oldest form of therapeutic treatment worldwide, and more than 21,000 plant species are used as herbal medicines according to the World Health Organization (Efferth, 2010). In particular, phytotherapy is practiced by the majority of the Mexican population for the treatment of many diseases. To promote the proper use of herbal medicines and to determine their potential use as a source of new drugs, it is essential to study medicinal plants and to scientifically validate their usage (Alonso-Castro et al., 2011). Natural products have proven to be an important source of lead compounds in the development of new drugs. Artemisinin, quinine and licochalcone A are examples of plant-derived products with antiparasitic activity. Screening natural products provides the chance to discover new molecules of unique structure with high activity and selectivity (Kayser et al., 2003). New therapeutic approaches have been developed for the treatment of Chagas disease that are based on natural plant products as an alternative source of drugs to combat T. cruzi infection, some of which exhibit trypanocidal activity and lower toxicity (Luize et al., 2005; Sülsen et al., 2006; Rojas et al., 2010). Despite the enormous variety of higher plant species, their potential as new drug sources has not been fully explored. Only 15–17% of this plant group has been systematically studied in the discovery of biologically active substances (Mafezoli et al., 2000; Adams et al., 2013). However, the larger portion of existing drugs has been derived from natural compounds, including semi-synthetic and synthetic derivatives based on natural product models (Newman and Cragg, 2012). As part of our ongoing study of trypanocidal constituents in plants, we have proceeded with the screening of the trypanocidal activity of some medicinal plants. The aim of the present work was to assess the in vitro trypanocidal activity of 10 plants used in Mexican folk medicine against T. cruzi via testing methanolic extracts of Artemisia mexicana, Castela texana, Cymbopogon citratus, Eryngium heterophyllum, Haematoxylum brasiletto, Lippia graveolens, Marrubium vulgare, Persea americana, Ruta chalepensis and Schinus molle. The selection of these plants was based on ethnomedicinal reports describing their use in the northeastern region of Mexico for the treatment of several diseases related to parasitic infections. Extracts of these plants have been commonly used as traditional medicines to treat bacterial, fungal and protozoan infections: A. mexicana is used to treat stomachache, diarrhea, parasitism and intestinal infections (Navarro et al., 1996); C. texana is used to treat amebic dysentery (Calzado-Flores et al., 2002); C. citratus is used to treat gastrointestinal diseases (Monzote et al., 2012); E. heterophyllum is used to treat diarrhea, stomachache, fever and cholelithiasis (BDMTM, 2013; Camou-Guerrero et al., 2008); H. brasiletto is used ˜ et al., 2012), L. graveto treat bacterial infections (Rosas-Pinón olens is used to treat gastrointestinal disorders, dysentery and giardiasis (Monzote et al., 2012); M. vulgare is used as hypotensive agent (Bardai et al., 2001); P. americana is used to treat fungal infections (Wang et al., 2004) and as a nematicidal agent (Dang et al., 2010); R. chalepensis is used as an antihelmintic and spasmolytic agent (Günaydin and Savci, 2005); and S. molle is used as a hypotensive and antispasmodic agent (Yueqin et al., 2003).
2. Materials and methods
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2.1. Plant materials
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The plants used in this study were collected in the municipalities of Aramberri and Sabinas Hidalgo, Nuevo León, México. Voucher specimens were deposited at the herbarium of the Universidad Autónoma de Nuevo León (UANL): A. mexicana (025533), C. texana (025538), C. citratus (025542), E. heterophyllum (025544), H. brasiletto (025548), L. graveolens (025554), M. vulgare (025555), P. americana (025563), R. chalepensis (025579) and S. molle (025567).
2.2. Preparation of plant extracts After drying at room temperature, leaves and aerial parts of the plants (20 g) were crushed into a powder and extracted with methanol in a Soxhlet extractor apparatus for 40 h. The solvent was subsequently removed using a rotary evaporator, and the concentrated substance was dried and stored in sealed glass vials at 4 ◦ C for further analysis (Pérez-Castorena et al., 2006; QuintanillaLicea et al., 2012). The following extract yields were obtained (g of extract/100 g plant): A. mexicana, 15.3%; C. texana, 14.0%; C. citratus, 17.6%; E. heterophyllum, 15.5%; H. brasiletto, 18.8%; L. graveolens, 41.0%; M. vulgare, 15.6%; P. americana, 21.2%; R. chalepensis, 12.7%; and S. molle, 15.9%.
2.3. Trypanocidal activity of plant extracts The trypanocidal activity of the plant extracts was assayed in the epimastigote form of T. cruzi (CL Brener strain). In view of the different responses to crude plant extracts, depending on the stages of the life cycle of T. cruzi, we decided to work only with the epimastigote stage because the positive correlation between activity against epimastigotes in vitro and activity against tripomastigotes in vivo has already been reported (Castro et al., 1992; Abe et al., 2002; Dantas et al., 2006). The parasites were cultivated in liver infusion tryptose (LIT) medium supplemented with 10% fetal bovine serum (FBS) and harvested during the exponential growth phase, when the parasites had reached a cell density of 2 × 106 epimastigotes/ml (Valencia et al., 2011). Stock solutions of each extract were prepared in 1% dimethyl sulfoxide (DMSO), which was a concentration that did not affect the growth of the parasites (Luize et al., 2005; Rojas et al., 2010). The bioassays were performed in duplicate in 96-well microtiter plates containing 200 l of the parasite suspension/well (Pizzolatti et al., 2002) and different concentrations (4.68–150 g/ml) of the extracts. Negative and positive controls containing epimastigotes in either DMSO (1%) or 10 g/ml of nifurtimox (Sigma-Aldrich, St. Louis, MO) were simultaneously performed (Muelas-Serrano et al., 2000). The assay plates were subsequently incubated for 96 h at 27 ◦ C (Pizzolatti et al., 2002; Luize et al., 2005), and the activity of each extract was categorized as low, moderate or high. The inhibitory effect on cell growth was estimated via the colorimetric tetrazolium dye (MTT) method (Muelas-Serrano et al., 2000; Machado et al., 2010). The concentrations inhibiting culture growth were determined through a dose–response regression analysis to obtain IC50 values (the concentration required for 50% inhibition, Santos et al., 2012), as plotted by the Prism 6 program (GraphPad Software, Inc., San Diego, CA). Trypanocidal activity was expressed as the percentage of T. cruzi epimastigotes (±SD) that were lysed (Pizzolatti et al., 2002).
Please cite this article in press as: Molina-Garza, Z.J., et al., Anti-Trypanosoma cruzi activity of 10 medicinal plants used in northeast Mexico. Acta Trop. (2014), http://dx.doi.org/10.1016/j.actatropica.2014.04.006
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Table 1 In vitro trypanocidal activity of methanolic extracts of the selected medicinal plants. Plant species
Part studied
% Growth inhibition ± SD 150 g/ml
Artemisia mexicana Castela texana Cymbopogon citrates Eryngium heterophyllum Haematoxylum brasiletto Lippia graveolens Marrubium vulgare Persea americana Ruta chalepensis Schinus molle Control (Nifurtimox)
AP L AP AP B L AP L L AP
83.93 32.44 83.30 88.38 93.04 33.01 98.99 61.32 75.85 100 80
± ± ± ± ± ± ± ± ± ± ±
0.03 4.06 0.70 0.11 6.9E−4 0.31 0.13 0.45 0.28 0.0 0.0
IC50 (g/ml) (95% CI) 39.25 (8.56–77.07) >150 68.25 (38.36–68.25) 11.24 (6.78–17.40) 7.92 (5.51–10.72) >150 22.66 (17.72–27.06) 65.51 (26.43–294.50) 72.30 (52.38–96.81) 16.31 (12.98–20.35) 10
AP, aerial part; L, leaves; B, bark; CI, confidence intervals.
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3. Results and discussion The in vitro trypanocidal activities of 10 medicinal plants used in northeast Mexico were evaluated. The activities of their methanolic extracts regarding inhibiting the growth of T. cruzi epimastigotes are shown in Table 1. The methanolic extracts of E. heterophyllum, H. brasiletto, M. vulgare and S. molle exhibited the highest activity against T. cruzi, achieving levels of growth inhibition between 88 and 100%. C. citratus and A. mexicana also exhibited good inhibitory activity, with approximately 83% inhibition of epimastigote growth observed under the analyzed concentrations. The P. americana and R. chalepensis extracts displayed a moderate level of activity against T. cruzi epimastigotes, resulting in approximately 70% inhibition. Finally, the methanolic extracts of C. texana and L. graveolens exhibited low activity against the protozoans (ca. 33% growth inhibition). Similar to previous reports, the positive control (nifurtimox) caused 80% growth inhibition at a concentration of 10 g/ml (MuelasSerrano et al., 2000). The methanolic extracts of H. brasiletto and E. heterophyllum showed the highest inhibitory effects against T cruzi at the lowest concentration (IC50 = 7.92 and 11.24 g/ml, respectively). Previous studies have shown that the crude methanolic extract of H. brasiletto can inhibit 100% of T. cruzi growth at 2 mg/ml and 80 to 90% of T. cruzi growth at 1 mg/ml (Abe et al., 2002). For E. heterophyllum, there are no previous studies of trypanocidal activity; however, some species of Eryngium, such as E. carlinae, have previously been reported to present trypanocidal activity (Abe et al., 2005). Additionally, crude extracts of E. amoriginum and E. ternatum have been tested for activity against Leishmania donovani and have shown inhibitory effects, displaying IC50 values between 2.0 and 9.5 g/ml, respectively (Fokialakis et al., 2007). S. molle also demonstrated good activity against T. cruzi (IC50 = 16.31 g/ml) in contrast to the DL50 of 50 g/ml in trypomastigote stage reported by Pizzolatti et al. (2002). This difference may be caused by different concentrations of the bioactive compounds in plants from distinct geographical regions. Otherwise, the essential oil of S. molle possesses trypanocidal activity over T. evansi (Baldissera et al., 2013), T. cruzi, T. brucei, and L. infantum, with an IC50 between 2 and 4.4 g/ml (Abdel-Sattar et al., 2010). In addition, leaf and fruit extracts of S. molle show repellent and insecticidal properties against nymphs and eggs of Triatoma infestans, which are vectors of Chagas disease (Ferrero et al., 2006). The strong inhibitory effects of M. vulgare (presenting IC50 = 22.66 g/ml) on the growth of epimastigotes in this study were of particular interest. In previous reports, the hexane fraction of M. vulgare showed stronger activity than the methanol fraction against T. cruzi epimastigotes (Gohari et al., 2008) and trypomastigotes in vitro (Abe et al., 2005). In addition, Abdel-Sattar et al. (2010) reported high inhibitory effects on the growth of T. brucei (IC50 = 0.25 g/ml) and L. infantum (IC50 = 2.03 g/ml).
We observed an acceptable level of activity in the extract of A. mexicana, which presented an IC50 = 39.25 g/ml. Other studies have shown that A. ludoviciana ssp. mexicana, which is a botanical synonym of A. mexicana, had a growth inhibitory effects of 80–90% at 2 mg/ml on the epimastigote stage of T. cruzi (Abe et al., 2002), which is less active than our extract. In addition, A. indica exhibits an excellent anti-leishmanicidal activity in all Leishmania strains studied, with an IC50 of 0.21 to 0.58 g/ml (Ganguly et al., 2006); and A. vulgari showed a high inhibition level of promastigote culture forms (Kheiri-Manjili et al., 2012). In the present study, the methanolic extract of C. citratus inhibited the growth of epimastigote forms of T. cruzi, presenting an IC50 = 68.25 g/ml, which is slightly higher than the IC50 (63.09 g/ml) observed for the essential oil of C. citratus against epimastigote forms of T. cruzi (Rojas et al., 2010). The same results were obtained with hydroalcoholic extracts of C. citratus, which confirmed that Leishmania species appear to be more sensitive to these extracts than T. cruzi (Luize et al., 2005; Machado et al., 2010). The essential oil of C. citratus also effectively inhibits the growth of L. chagasi (Oliveira et al., 2009). P. americana and R. chalepensis showed growth inhibition at concentrations of 65.51 and 72.30 g/ml (IC50 ), respectively. Moreover, similar results have previously demonstrated the moderate activity of methanolic extracts of the seeds of P. americana against epimastigotes (Abe et al., 2005). For R. chalepensis, there are no previous studies of trypanocidal activity; however, the onset of molting is delayed in Rhodnius prolixus, which is a vector of Chagas disease, when exposed to the odor of R. chalepensis (Abramson et al., 2007). In the present study, the extracts of C. texana and L. graveolens were inactive against T. cruzi epimastigotes. In similar studies, C. texana also showed no activity against epimastigotes (Abe et al., 2005), and hydroalcoholic extracts of L. alba were found to be inactive against L. amazonensis and T. cruzi (Luize et al., 2005). There are reports concerning the role of some plant chemical compositions in the trypanocidal activity of pure compounds isolated from some of the studied species of plants in this current study, including hematoxylin, brazilin, caffeic acid, methyl gallate, gallic acid, phloroglucinol, 4-hydroxycinnamic acid and 5-methoxypsoralen, which were isolated from H. brasiletto (RiveroCruz, 2008). Commercially acquired gallic acid, with an IC50 of 67.0 g/ml (Tasdemir et al., 2006), and 4-hydroxycinnamic acid derivatives isolated from Brazilian propolis (Salomão et al., 2008), showed trypanocidal activity against T. cruzi. Neither chemical nor biological reports concerning the trypanocidal activity of E. heterophyllum could be found in the literature; however, (E)-2-dodecenal isolated from E. foetidum possesses important trypanocidal activity (Forbes and Steglich, 2007). Phytochemical studies of S. molle showed the presence of ␣phellandrene, -phellandrene, ␣-terpineol, ␣-pinene, -pinene and p-cymene (Bendaoud et al., 2010), as well as -caryophyllene, ␣-copaene and ␣-humulene (Díaz et al., 2008; Deveci et al., 2010).
Please cite this article in press as: Molina-Garza, Z.J., et al., Anti-Trypanosoma cruzi activity of 10 medicinal plants used in northeast Mexico. Acta Trop. (2014), http://dx.doi.org/10.1016/j.actatropica.2014.04.006
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The essential oils of Annona pickelii and Annona salzmannii, which contain -caryophyllene, ␣-copaene and ␣-humulene, showed an IC50 lower than 100 g/ml against the trypomastigote stage of T. cruzi (Costa et al., 2013). In a similar manner, caryophylene derivatives present in the essential oil of Hedychium coronarium showed trypanocidal activity against T. brucei (Costa et al., 2013; Rodrigues et al., 2013). From M. vulgare, numerous terpenes, such as ␥-eudesmol, citronellol, citronellyl formate and germacrene D (Kadril et al., 2011), and some flavonoids, such as apigenin, luteolin and vitexin, were isolated (Nawwar et al., 1989). Commercially acquired flavonoids and derivatives, which were found in M. vulgare, possess important trypanocidal activity. This situation is the case for luteolin, with an IC50 of 21.4 g/ml, for luteolin-7-O-glucoside, vitexin and apigenin-7-O-glucoside, with a major IC50 of 30 g/ml, and, finally, apigenin, with an IC50 = 21.8 g/ml (Tasdemir et al., 2006). From A. mexicana, caffeic acid, gallic acid, kampferol, quercetin, myricetin and -caryophyllene (Lopes-Lutz et al., 2008; Carvalho et al., 2011) have been isolated, and there are reports that the commercially acquired compounds possess important trypanocidal activity (Tasdemir et al., 2006; Rodrigues et al., 2013). Citral, which is a major compound found in C. citratus, possesses inhibitory effects in vitro over T. cruzi at a concentration of 40 g/ml at 24 h after application during the differentiation process (Cardoso and Soares, 2010) and kills 65% of L. infantum and L. major and 80% of L. tropica promastigotes at a concentration of 50 g/ml (Machado et al., 2010, 2012). 1,2,4-Trihydroxyheptadecane and 1,2,4-trihydroxynonadecane derivatives obtained from seeds of P. americana exhibit moderate activity against epimastigotes and trypomastigotes of T. cruzi (Abe et al., 2005). Several furanocoumarines and quinolone alkaloids were isolated from R. chalepensis (El-Sayed et al., 2000; Günaydin and Savci, 2005). Some commercially acquired furanocoumarins, such as umbelliferone and bergapten exhibit moderate activity against epimastigotes and trypomastigotes of T. cruzi (Tasdemir et al., 2006). In recent years, several studies have used medicinal plants for the treatment of diseases related to infections with protozoa and with other microorganisms. Cytotoxic activities of crude extracts of various plant species have been demonstrated against amastigotes, epimastigotes and trypomastigotes of T. cruzi (Mafezoli et al., 2000; Pizzolatti et al., 2002; Santoro et al., 2007). Additionally, although the reproducibility of the growth inhibition of T. cruzi culture forms in vitro and in vivo has been previously demonstrated (Schlemper et al., 1977; Abe et al., 2002), some studies have shown that plant extracts exhibit higher trypanocidal activity against some T. cruzi stages in vitro or in vivo (Abe et al., 2005; Castro et al., 1992; Dantas et al., 2006). We consider that advantages of the screening method with T. cruzi epimastigotes include establishing fast and reliable anti-T. cruzi screening programs at low costs, despite the lack of large animal facilities, and the availability of low amounts of drugs or crude extracts. In addition, the manipulation of culture forms is most likely less risky than the handling of infected animals (Schlemper et al., 1977). Several natural products, such as alkaloids, terpenes and quinones, flavonoids and saponins, with trypanocidal activity have been isolated (Wright and Phillipson, 1990; Kayser et al., 2003). Considering these findings, we can conclude that the results obtained in this study indicate that the most promising extracts may result in potential sources of lead compounds for the development of more effective drugs for the treatment of Chagas disease. The bioguided fractionation of E. heterophyllum, H. brasiletto, M. vulgare and S. molle active extracts is ongoing, and further studies of the isolated pure compounds in vivo with amastigotes and trypomastigotes stages of T. cruzi will be performed. Furthermore, these
results suggest that a variety of potentially bioactive substances exist in the Mexican flora.
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We gratefully acknowledge the contributions of QBP Paola Katyana Vázquez-Luévanos and QBP Diana Janeth Coronado˜ who performed the laboratory work. In addition, special Zermeno, thanks go to Dr. José Luis Rosales Encina and Biol. Lidia Baylon (Departamento de Infectómica y Patogénesis Molecular, Centro de Investigación y de Estudios Avanzados, México), who kindly provided us with the CL Brener strain. This project was partially supported by PAICYT-UANL CN835-11. Q2 Q3
References Abdel-Sattar, E., Maes, L., Salama, M.M., 2010. In vitro activities of plant extracts from Saudi Arabia against malaria, leishmaniasis, sleeping sickness and Chagas disease. Phytother. Res. 24, 1322–1328. Adams, M., Chammartin, M., Hamburger, M., Potterat, O., 2013. Case study of the Swiss flora for prior phytochemical and biological investigations. J. Nat. Prod. 76, 209–215. Abe, F., Nagafuji, S., Yamauchi, T., Okabe, H., Maki, J., Higo, H., Akahane, H., Aguilar, A., Jiménez-Estrada, M., Reyes-Chilpa, R., 2002. Trypanocidal constituents in plants. 1. Evaluation of some Mexican plants for their trypanocidal activity and active constituents in Guaco, roots of Aristolochia taliscana. Biol. Pharm. Bull. 25, 1118–1191. Abe, F., Nagafuji, S., Okawa, M., Kinjo, J., Akahane, H., Ogura, T., Martinez-Alfaro, M.A., Reyes-Chilpa, R., 2005. Trypanocidal constituents in plants. 5. Evaluation of some Mexican plants their trypanocidal activity and active constituents in the seeds of Persea americana. Biol. Pharm. Bull. 28, 1314–1317. Abramson, C.H.I., Aldana, E., Sulbaran, E., 2007. Exposure to citral, cinnamon and ruda disrupts the life cycle of a vector of Chagas disease. Am. J. Environ. Sci. 3, 7–8. Alonso-Castro, A.J., Villarreal, M.L., Salazar-Olivo, L.A., Gomez-Sanchez, M., Dominguez, F., Garcia-Carranca, A., 2011. Mexican medicinal plants used for cancer treatment: pharmacological, phytochemical and ethnobotanical studies. J. Ethnopharmacol. 133, 945–972. Baldissera, M.D., Da Silva, A.S., Oliveira, C.B., Zimmermann, C.E.P., Vaucher, R.A., Santos, R.C.V., Rech, V.C., Tonin, A.A., Giongo, J.L., Mattos, C.B., Koester, L., Santuario, J.M., Monteiro, S.G., 2013. Trypanocidal activity of the essential oils in their conventional and nanoemulsion forms: in vitro tests. Exp. Parasitol. 134, 356–361. Bardai, S.E., Lyoussi, B., Wibo, M., Morel, N., 2001. Pharmacological evidence of hypotensive activity of Marrubium vulgare and Foeniculum vulgare in spontaneously hypertensive rat. Clin. Exp. Hypertens. 23, 329–343. Bastos, C.J.C., Aras, R., Mota, G., Reis, F., Pereira Dias, J., Silva de Jesus, R., Silva Freire, M., de Araújo, E.G., Prazeres, J., Rios Grassi, M.F., 2010. Clinical outcomes of thirteen patients with acute Chagas disease acquired through oral transmission from two urban outbreaks in northeastern Brazil. PLoS Negl.Trop. Dis. 4, e711. BDMTM (Biblioteca Digital de la Medicina Tradicional Mexicana), 2013. Eryngium heterophyllum. BDMTM, . Bendaoud, H., Romdhone, M., Souchard, J.P., Cazaux, S., Bouajila, J., 2010. Chemical composition and anticancer and antioxidant activities of Schinus molle L. and Schinus terebinthifolius raddi berries essential oil. J. Food Sci. 75, 466–472. Bern, C., Kjos, S., Yabsley, M.J., Montgomery, S.P., 2011. Trypanosoma cruzi and Chagas’ disease in the United States. Clin. Microbiol. Rev. 24, 655–681. Calzado-Flores, C., Hurtado Ramírez, M.B., Flores Villanueva, Z., Verde Star, M.J., Segura Luna, J.J., 2002. Preliminary morphological study of the effects produced by a methanolic extract of Castela texana. Proc. West. Pharmacol. Soc. 45, 161. Camou-Guerrero, A., Reyes-García, V., Martínez-Ramos, M., Casas, A., 2008. Knowledge and use value of plant species in a Rarámuri community: a gender perspective for conservation. Hum. Ecol. 36, 259–272. Carabarin-Lima, A., González-Vázquez, M.C., Rodríguez-Moralesa, O., BaylónPacheco, L., Rosales-Encina, J.L., Reyes-López, P.A., Arce-Fonseca, M., 2013. Chagas disease (American trypanosomiasis) in Mexico: an update. Acta Trop. 127, 126–135. Cardenas-Sánchez, J.D., Mazariego-Arana, M.A., Utrilla-Pascacio, F.J., MonteónPadilla, V.M., Altúzar-González, M., 2003. Anticuerpos anti-Trypanosoma cruzi en pacientes con cardiomiopatía dilatada. Rev. Med. Mex. 41, 111–114. Cardoso, J., Soares, M.J., 2010. In vitro effects of citral on Trypanosoma cruzi metacyclogenesis. Mem. Inst. Oswaldo Cruz 105, 1026–1032. Carod-Artal, F.J., Vargas, A.P., Horan, T.A., Nadal-Nunes, L.G., 2005. Chagasic cardiomyopathy is independently associated with ischemic stroke in Chagas disease. Stroke 36, 965–970. Carvalho, I.S., Cavaco, T., Brodelius, M., 2011. Phenolic composition and antioxidant capacity of six artemisia species. Ind. Crops Prod. 33, 382–388. Castro, S.L., Soeiro, M.N.C., Meirelles, M.N.L., 1992. Trypanosoma cruzi: effect of phenothiazines on the parasite and its interactions with host cells. Mem. Inst. Oswaldo Cruz 87, 209–215.
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427 428 429 430 431 432 433 434 435 436 437 438 439 440 441 442 443 444 445 446 447 448 449 450 451 452 453 454 455 456 457 458 459 460 461 462 463 464 465 466 467 468 469 470 471 472 473 474 475 476 477 478 479 480 481 482 483 484 485 486 487 488 489 490 491 492 493 494 495 496 497 498 499 500 501 502 503 504 505 506 507 508 509 510
Chagas, C., 1909. Nova tripanozomiase humana: estudos sobre a morfolojia e o ciclo evolutivo do Schizotrypanum cruzi n. gen., n. sp., ajente etiolojico de nova entidade mórbida do homem. Mem. Inst. Oswaldo Cruz 1, 159–218. Costa, E.U., Dutra, L.M., Salvador, M.J., Gunzaga, R.L.H., Gadelha, R.F., Carvalho, J.E., 2013. Chemical composition of the essential oils of Annona pickelii and Annona salzmannii (Annonaceae) and their antitumour and trypanocidal activities. Nat. Prod. Res. 27, 997–1001. Cruz-Reyes, A., Pickering-Lopez, J.M., 2006. Chagas disease in Mexico: an analysis of geographical distribution during the past 76 years—a review. Mem. Inst. Oswaldo Cruz 101, 345. Dang, Q.L., Kwon, H.R., Choi, Y.H., Choi, G.J., Jang, K.S., Park, M.S., Lim, C.H., Ngoc, L.H., Kim, J.C., 2010. Nematicidal activity against Bursaphelenchus xylophilus of isoobtusilactone A isolated from Persea americana. Nematology 12, 247–253. Dantas, A.P., Salomão, K., Barbosa, H.S., De Castro, S.L., 2006. The effect of Bulgarian propolis against Trypanosoma cruzi and during its interaction with host cells. Mem. Inst. Oswaldo Cruz 101, 207–211. Deveci, O., Sukan, A., Tuzun, N., Kocabas, E.E.H., 2010. Chemical composition, repellent and antimicrobial activity of Schinus molle L. J. Med. Plants Res. 4, 2211–2216. Díaz, C., Quesada, S., Brenes, O., Aguilar, G., Cicció, F.J., 2008. Chemical composition of Schinus molle essential oil and its citotoxic activity on tumour cell lines. Nat. Prod. Res. 22, 1521–1534. El-Sayed, K., Al-Said, M.S., El-Feraly, F.S., Ross, S.A., 2000. New quinoline alkaloids from Ruta chalepensis. J. Nat. Prod. 63, 995–997. Efferth, T., 2010. Cancer therapy with natural products and medicinal plants. Planta Med. 76, 1035–1036. Ferrero, A.A., Werdin-González, J.O., Sánchez-Chopa, C., 2006. Biological activity of Schinus molle on Triatoma infestans. Fitoterapia 77, 381–383. Fokialakis, N., Kalpoutzakis, E., Tekwani, B.L., Khan, S.I., Kobaisy, M., Skaltsounis, A.L., Duke, S.O., 2007. Evaluation of the antimalarial and antileishmanial activity of plants from the Greek island of Crete. J. Nat. Med. 61, 38–45. Forbes, W.M., Steglich, C., 2007. Methods of treating infectious diseases. In: United States Patent Application 20090047342. Slippery Rock University (Assignee), Slippery Rock, PA) (filed 08/15). Galaviz-Silva, L., Molina-Garza, D.P., González-Santos, M.A., Mercado-Hernández, R., González-Galaviz, J.R., Rosales-Encina, J.L., Molina-Garza, Z.J., 2009. Update on seroprevalence of anti-Trypanosoma cruzi antibodies among blood donors in northeast Mexico. Am. J. Trop. Med. Hyg. 81, 404–406. Ganguly, S., Bandyopadhyay, S., Bera, A., Chatterjee, M., 2006. Antipromastigote activity of an ethanolic extract of leaves Artemisa indica. Indian J. Pharmacol. 38, 64–65. Gohari, A.R., Saeidnia, S., Hadjiakhoondi, A., Naghinejad, A., Yagura, T., 2008. Trypanocidal activity of some medicinal plants against the epimastigotes of Trypanosoma cruzi. J. Med. Plants 7, 44–48. Günaydin, K., Savci, S., 2005. Phytochemical studies on Ruta chalepensis (Lam.) Lamark. Nat. Prod. Res. 19, 203–210. Gürtler, R.E., Segura, E.L., Cohen, J.E., 2003. Congenital transmission of Trypanosoma cruzi infection in Argentina. Emerg. Infect. Dis. 9, 29–32. Kadril, A., Zarai, Z., Békir, A., Gharsallah, N., Damak, M., Gdoura, R., 2011. Chemical composition and antioxidant activity of Marrubium vugare L. essential oil from Tunisia. Afr. J. Biotech. 10, 3908–3914. Kayser, O., Kiderlen, A.F., Croft, S.I., 2003. Natural products as antiparasitic drugs. Parasitol. Res. 90, S55–S62. Kheiri-Manjili, H., Jafari, H., Ramazani, A., Davoudi, N., 2012. Anti-leishmanial and toxicity activities of some selected Iranian medicinal plants. Parasitol. Res. 111, 2115–2121. Kirchhoff, L.V., Paredes, P., Lomelí-Guerrero, A., Paredes-Espinoza, M., Ron-Guerrero, ˜ ˜ noz, J.G., 2006. Transfusion-associated ChaC.S., Delgado-Mejía, M., Pena-Mu gas disease (American tripanosomiasis) in Mexico: implications for transfusion medicine in the United States. Transfusion 46, 298–304. Lopes-Lutz, D., Alviano, D.S., Alviano, C.S., Kolodziejczyk, P.P., 2008. Screening of chemical composition, antimicrobial and antioxidant activities of artemisia essential oils. Phytochemistry 69, 1732–1738. Luize, P.S., Shioji-Tiuman, T., Morello, L.G., Korehiza Maza, P., 2005. Effects of medicinal plant extracts on growth of Leishmania (L.) amazonensis and Trypanosoma cruzi. Braz. J. Pharm. Sci. 41, 85–94. Machado, M., Santoro, G., Sousa, M.C., Salgueiro, L., Cavaleiro, C., 2010. Activity of essential oils on the growth of Leishmania infantum promastigotes. Flavour Fragance J. 25, 156–160. Machado, M., Pires, P., Dinis, A.M., Santos-Rosa, M., Alves, V., Salgueiro, L., Cavaleiro, C., Sousa, M.C., 2012. Aldheides monoterpenes as potential anti-Leishmania agents: activity of Cymbopongon citratus and Citral on L. infantum, L. tropica and L. major. Exp. Parasitol. 130, 223–231. Mafezoli, J., Vieira, P.C., Fernandes, J.B., da Silva, M.F.G.F., de Albuquerque, S., 2000. In vitro activity of Rutaceae species against the trypomastigote form of Trypanonosma cruzi. J. Ethnopharmacol. 73, 335–340. Martínez-Ibarra, A., Galaviz-Silva, L., Trujilla, C., 1992. Distribución de los triatominos asociados al domicilio humano en el municipio de General Terán, Nuevo León, México. Southwest. Entomol. 17, 261–265. Molina-Garza, Z.J., Galaviz-Silva, L., Molina-Garza, D., Rosales-Encina, J.L., 2007. Prevalencia de Trypanosoma cruzi en triatominos de Nuevo León, México. Salud Pública Mex. 49, 37–44. Monzote, L., Alarcón, O., Setzer, W.N., 2012. Antiprotozoal activity of essential oils review. Agric. Conspec. Sci. 77, 167–175.
5
Muelas-Serrano, S., Nogal-Ruiz, J., Gómez-Barrio, A., 2000. Setting of a colorimetric method to determine the viability of Trypanosoma cruzi epimastigotes. Parasitol. Res. 86, 999–1002. Navarro, V., Villarreal, M.L., Rojas, G., Lozoya, X., 1996. Antimicrobial evaluation of some plants used in Mexican traditional medicine for the treatment of infectious diseases. J. Ethnopharmacol. 53, 143–147. Nawwar, M.A.M., El-monsallamy, A.M.D., Barakat, H.H., Buddrus, J., Linscheid, M., 1989. Flavonoid lactates from leaves of Marrubium vulgare. Phytochemistry 28, 3201–3206. Newman, D.J., Cragg, G.M., 2012. Natural products as sources of new drugs over the 30 years from 1981 to 2010. J. Org. Chem. 75, 311–335. PAHO (Pan American Health Organization), 2013. Infographic: Chagas in the Americas. Factsheet-chagas-eng-2013. PAHO, . Pérez-Castorena, A.L., Oropeza, R.F., Vázquez, A.R., Martínez, M., Maldonado, E., 2006. Labdanes and withanolides from physalis coztomatl. J. Nat. Prod. 69, 1029–1033. Pizzolatti, M.G., Koga, A.H., Grisard, E.C., Steindel, M., 2002. Trypanocidal activity of extracts from Brazilian Atlantic rain forest plant species. Phytomedicine 9, 422–426. Quintanilla-Licea, R., Morado-Castillo, R., Gomez-Flores, R., Laatsch, H., Verde-Star, M.J., Hernández-Martínez, H., Tamez-Guerra, P., Tamez-Guerra, R., RodríguezPadilla, C., 2012. Bioassay-guided isolation and identification of cytotoxic compounds from Gymnosperma glutinosum leaves. Molecules 17, 11229–11241. Oliveira, V.C.S., Moura, D.M.S., Lopes, J.A.D., de Andrade, P.P., da Silva, N.H., Figueiredo, R.C.B.Q., 2009. Effects of essential oils from Cymbopogon citratus (DC) Stapf., Lippia sidoides Cham., and Ocimum gratissimum L. on growth and ultrastructure of Leishmania chagasi promastigotes. Parasitol. Res. 104, 1053–1059. Reisenman, C.E., Lawrence, G., Guerenstein, P.G., Gregory, T., Dotson, E., Hildebrand, J.G., 2010. Infection of kissing bugs with Trypanosoma cruzi, Tucson Arizona, USA. Emerg. Infect. Dis. 16, 400–405. Rivero-Cruz, J.F., 2008. Antimicrobial compounds isolated from Haematoxylon brasiletto. J. Ethnopharmacol. 119, 99–103. Rodrigues, D.F., Arenas, V.A.M., Cavaleiro, C., Salgueiro, L., Zimmermann, M.G., Oliveira, M.N., Guncalves, M.M.B., Barretto, C.R.M., Duarte, M.R.R., 2013. Chemical composition and trypanocidal activity of the essential oils from Hedychium coronarium J. Koening (Zingiberaceae). ISRN Infect. Dis. 2013, 1–6. Rojas, J., Solís, H., Palacios, O., 2010. Evaluación in vitro de la actividad anti Trypanosoma cruzi de aceites esenciales de diez plantas medicinales. Anal. Facult. Med. 71, 161–165. ˜ Y., Mejía, A., Díaz-Ruiz, G., Aguilar, M.S., Sánchez-Nieto, S., RiveroRosas-Pinón, Cruz, J.F., 2012. Ethnobotanical survey and antibacterial activity of plants used in the Altiplane region of Mexico for the treatment of oral cavity infections. J. Ethnopharmacol. 141, 860–865. Salomão, K., Pereira, S.P.R., Campos, L.C., Borba, C.M., Cabello, P.H., Marcucci, M.C., de Castro, S.L., 2008. Brazilian Propolis: correlation between chemical composition and antimicrobial anctivity. eCAM. , 317–324. Santoro, G.F., Cardoso, M.G., Guimarães, L.G., Freire, J.M., Soares, M.J., 2007. Anti-proliferative effect of essential oil of Cymbopogon citratus (DC) Stapf (Lemongrass) on intracellular amastigotes, bloodstream trypomastigotes and culture epimastigotes of Trypanosoma cruzi (Protozoa:Kinetoplastida). Parasitology 134, 1649–1656. Santos, K,K,A., Matias, E.F.F., Tintino, S.R., Souza, C.E.S., Braga, M.F.B., Guedes, G.M.M., Rolón, M., Vega, C., de Arias, A.R., Costa, J.G.M., Menezes, I.R.A., Coutinho, H.D.M., 2012. Anti-Trypanosoma cruzi and cytotoxic activities of Eugenia uniflora L. Exp. Parasitol. 131, 130–132. Schlemper, B.R., Chiari, E., Brener, Z., 1977. Growth-inhibition drug test with Trypanosoma cruzi culture forms. J. Protozool. 24, 544–547. Schmunis, G.A., 2007. Epidemiology of Chagas disease in non-endemic countries: the role of international migration. Mem. Inst. Oswaldo Cruz. 102, 75–85. Sülsen, V., Güida, C., Coussio, J., Paveto, C., Muschietti, L., Martino, V., 2006. In vitro evaluation of trypanocidal activity in plants used in Argentine traditional medicine. Parasitol. Res. 98, 370–374. Tasdemir, D., Kaiser, M., Brun, R., Yardley, V., Schmidt, T.J., Tosun, F., Rüedi, P., 2006. Antitrypanosomal and antileishmanial activities of flavonoids and their analogues: in vitro, in vivo, structure activity relationship studies. Antimicrob. Agents Chemother. 50, 1352–1364. Troullier, P., Olliaro, P., Torreele, E., Orbinski, J., Laing, R., Ford, N., 2002. Drug development for neglected diseases: a deficient market and a public health policy failure. Lancet 359, 2188–2194. ˜ D.L., Robledo, S.M., Echeverri, F., Arango, G.J., Vélez, I.D., Triana, Valencia, L., Munoz, O., 2011. Actividad tripanocida y citotóxica de extractos de plantas colombianas. Biomédica 31, 552–559. Wang, X., Beno-Movalem, D., Kobiler, I., Leikin-Frenkel, A., Lichter, A., Prusky, D., 2004. Expression of 12 fatty acid desaturase during the induced accumulation of the antifungal diene in avocado fruits. Mol. Plant Pathol. 5, 575–585. WHO (World Health Organization), 2014. Chagas disease (American trypanosomiasis). In: Fact Sheet No 340, , (updated March, 2013). Wright, C.W., Phillipson, J.D., 1990. Natural products and the development of selective antiprotozal drugs. Phytother. Res. 4, 127–139. ˜ S., Giner, R.M., Cerdá-Nicolás, M., Ríos, J.L., 2003. IsoYueqin, Z., Recio, M.C., Mánez, lation of two triterpenoids and a bioflavanone with anti-inflammatory activity from Schinus molle fruits. Planta Med. 69, 893–898.
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Anti-Trypanosoma cruzi activity of 10 medicinal plants used in northeast Mexico
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Zinnia Judith Molina-Garza a , Aldo Fabio Bazaldúa-Rodríguez b , Ramiro Quintanilla-Licea b , Lucio Galaviz-Silva a,∗ a Universidad Autónoma de Nuevo León, UANL, Facultad de Ciencias Biológicas, Laboratorio de Patología Molecular, Ave. Universidad S/N, Cd. Universitaria, 66451 San Nicolás de los Garza, NL, Mexico b Universidad Autónoma de Nuevo León, UANL, Facultad de Ciencias Biológicas, Laboratorio de Fitoquímica, Ave. Universidad S/N, Cd. Universitaria, 66451 San Nicolás de los Garza, NL, Mexico
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Article history: Received 20 November 2013 Received in revised form 3 April 2014 Accepted 5 April 2014 Available online xxx
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Keywords: Trypanocidal activity Chagas disease Medicinal plants
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1. Introduction
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The aim of this study was to screen the trypanocidal activity of plants used in traditional Mexican medicine for the treatment of various diseases related to parasitic infections. Cultured Trypanosoma cruzi epimastigotes were incubated for 96 h with different concentrations of methanolic extracts obtained from Artemisia mexicana, Castela texana, Cymbopogon citratus, Eryngium heterophyllum, Haematoxylum brasiletto, Lippia graveolens, Marrubium vulgare, Persea americana, Ruta chalepensis and Schinus molle. The inhibitory concentration (IC50 ) was determined for each extract via a colorimetric method. Among the evaluated species, the methanolic extracts of E. heterophyllum, H. brasiletto, M. vulgare and S. molle exhibited the highest trypanocidal activity, showing percentages of growth inhibition between 88 and 100% at a concentration of 150 g/ml. These medicinal plants may represent a valuable source of new bioactive compounds for the therapeutic treatment of trypanosomiasis. © 2014 Published by Elsevier B.V.
Chagas disease is caused by the protozoan parasite Trypanosoma cruzi, and over 100 years after its discovery by Carlos Chagas (Chagas, 1909), this disease continues to represent a major health issue in Latin America. Initially, this disease primarily occurred in rural areas, where the causative agent was transmitted from blood-sucking insects of the Reduviidae family to humans (Cardenas-Sánchez et al., 2003); however, currently, the accidental oral transmission of T. cruzi is becoming increasingly common (Bastos et al., 2010), whereas transmission by vectors and by blood transfusion has substantially decreased throughout Latin America (WHO, 2014). In addition, migration has brought infected individuals to urbanized areas of Latin America and to Europe, Japan, Australia (Schmunis, 2007) and the United States (Carod-Artal et al., 2005; Reisenman et al., 2010; Bern et al., 2011), where infections through non-vectorial routes can occur via blood
∗ Corresponding author. Tel.: +52 81 83524425/+52 83 524425. E-mail addresses: [email protected], [email protected] (L. Galaviz-Silva).
transfusion (Kirchhoff et al., 2006; Galaviz-Silva et al., 2009), organ transplantation, and congenital transmission (Gürtler et al., 2003; Schmunis, 2007). This disease is also known as American trypanosomiasis, and approximately 7–8 million people are currently infected (WHO, 2014). Furthermore, the estimated number of people infected worldwide has declined to 8 million, with an annual incidence rate of 56,000 cases and an estimated 12,000 deaths occurring every year (PAHO, 2013). Mexico is a country with high climatic variety and great biodiversity; this environment provides excellent habitats for the geographic distribution of Triatominae species, which can be found in most Mexican states (Cruz-Reyes and Pickering-Lopez, 2006). In the state of Nuevo León in northeast Mexico, the estimated population at risk of T. cruzi infection is approximately 90,277 individuals (Carabarin-Lima et al., 2013). Only Triatoma gerstaeckeri (Stål) has been characterized as a domiciliary and peridomestic vector, whereas T. neotomae (Neiva), T. lecticularia (Stål) and T. protracta (Uhler) are involved in sylvatic cycles (Martínez-Ibarra et al., 1992; Molina-Garza et al., 2007). Nifurtimox and benznidazole have been used for over 40 years to treat Chagas disease; however, these drugs are only effective during the acute phase of infection, and both pharmaceuticals induce
http://dx.doi.org/10.1016/j.actatropica.2014.04.006 0001-706X/© 2014 Published by Elsevier B.V.
Please cite this article in press as: Molina-Garza, Z.J., et al., Anti-Trypanosoma cruzi activity of 10 medicinal plants used in northeast Mexico. Acta Trop. (2014), http://dx.doi.org/10.1016/j.actatropica.2014.04.006
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significant side effects after long periods of medication usage. Furthermore, certain protozoan strains have developed resistance to treatment with these drugs (Sülsen et al., 2006; Rojas et al., 2010). The development of new, safer and more effective trypanocidal compounds remains a challenge because these drugs are not given high priority by the R&D-based pharmaceutical industry (Troullier et al., 2002; Sülsen et al., 2006). Phytotherapy represents the oldest form of therapeutic treatment worldwide, and more than 21,000 plant species are used as herbal medicines according to the World Health Organization (Efferth, 2010). In particular, phytotherapy is practiced by the majority of the Mexican population for the treatment of many diseases. To promote the proper use of herbal medicines and to determine their potential use as a source of new drugs, it is essential to study medicinal plants and to scientifically validate their usage (Alonso-Castro et al., 2011). Natural products have proven to be an important source of lead compounds in the development of new drugs. Artemisinin, quinine and licochalcone A are examples of plant-derived products with antiparasitic activity. Screening natural products provides the chance to discover new molecules of unique structure with high activity and selectivity (Kayser et al., 2003). New therapeutic approaches have been developed for the treatment of Chagas disease that are based on natural plant products as an alternative source of drugs to combat T. cruzi infection, some of which exhibit trypanocidal activity and lower toxicity (Luize et al., 2005; Sülsen et al., 2006; Rojas et al., 2010). Despite the enormous variety of higher plant species, their potential as new drug sources has not been fully explored. Only 15–17% of this plant group has been systematically studied in the discovery of biologically active substances (Mafezoli et al., 2000; Adams et al., 2013). However, the larger portion of existing drugs has been derived from natural compounds, including semi-synthetic and synthetic derivatives based on natural product models (Newman and Cragg, 2012). As part of our ongoing study of trypanocidal constituents in plants, we have proceeded with the screening of the trypanocidal activity of some medicinal plants. The aim of the present work was to assess the in vitro trypanocidal activity of 10 plants used in Mexican folk medicine against T. cruzi via testing methanolic extracts of Artemisia mexicana, Castela texana, Cymbopogon citratus, Eryngium heterophyllum, Haematoxylum brasiletto, Lippia graveolens, Marrubium vulgare, Persea americana, Ruta chalepensis and Schinus molle. The selection of these plants was based on ethnomedicinal reports describing their use in the northeastern region of Mexico for the treatment of several diseases related to parasitic infections. Extracts of these plants have been commonly used as traditional medicines to treat bacterial, fungal and protozoan infections: A. mexicana is used to treat stomachache, diarrhea, parasitism and intestinal infections (Navarro et al., 1996); C. texana is used to treat amebic dysentery (Calzado-Flores et al., 2002); C. citratus is used to treat gastrointestinal diseases (Monzote et al., 2012); E. heterophyllum is used to treat diarrhea, stomachache, fever and cholelithiasis (BDMTM, 2013; Camou-Guerrero et al., 2008); H. brasiletto is used ˜ et al., 2012), L. graveto treat bacterial infections (Rosas-Pinón olens is used to treat gastrointestinal disorders, dysentery and giardiasis (Monzote et al., 2012); M. vulgare is used as hypotensive agent (Bardai et al., 2001); P. americana is used to treat fungal infections (Wang et al., 2004) and as a nematicidal agent (Dang et al., 2010); R. chalepensis is used as an antihelmintic and spasmolytic agent (Günaydin and Savci, 2005); and S. molle is used as a hypotensive and antispasmodic agent (Yueqin et al., 2003).
2. Materials and methods
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2.1. Plant materials
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The plants used in this study were collected in the municipalities of Aramberri and Sabinas Hidalgo, Nuevo León, México. Voucher specimens were deposited at the herbarium of the Universidad Autónoma de Nuevo León (UANL): A. mexicana (025533), C. texana (025538), C. citratus (025542), E. heterophyllum (025544), H. brasiletto (025548), L. graveolens (025554), M. vulgare (025555), P. americana (025563), R. chalepensis (025579) and S. molle (025567).
2.2. Preparation of plant extracts After drying at room temperature, leaves and aerial parts of the plants (20 g) were crushed into a powder and extracted with methanol in a Soxhlet extractor apparatus for 40 h. The solvent was subsequently removed using a rotary evaporator, and the concentrated substance was dried and stored in sealed glass vials at 4 ◦ C for further analysis (Pérez-Castorena et al., 2006; QuintanillaLicea et al., 2012). The following extract yields were obtained (g of extract/100 g plant): A. mexicana, 15.3%; C. texana, 14.0%; C. citratus, 17.6%; E. heterophyllum, 15.5%; H. brasiletto, 18.8%; L. graveolens, 41.0%; M. vulgare, 15.6%; P. americana, 21.2%; R. chalepensis, 12.7%; and S. molle, 15.9%.
2.3. Trypanocidal activity of plant extracts The trypanocidal activity of the plant extracts was assayed in the epimastigote form of T. cruzi (CL Brener strain). In view of the different responses to crude plant extracts, depending on the stages of the life cycle of T. cruzi, we decided to work only with the epimastigote stage because the positive correlation between activity against epimastigotes in vitro and activity against tripomastigotes in vivo has already been reported (Castro et al., 1992; Abe et al., 2002; Dantas et al., 2006). The parasites were cultivated in liver infusion tryptose (LIT) medium supplemented with 10% fetal bovine serum (FBS) and harvested during the exponential growth phase, when the parasites had reached a cell density of 2 × 106 epimastigotes/ml (Valencia et al., 2011). Stock solutions of each extract were prepared in 1% dimethyl sulfoxide (DMSO), which was a concentration that did not affect the growth of the parasites (Luize et al., 2005; Rojas et al., 2010). The bioassays were performed in duplicate in 96-well microtiter plates containing 200 l of the parasite suspension/well (Pizzolatti et al., 2002) and different concentrations (4.68–150 g/ml) of the extracts. Negative and positive controls containing epimastigotes in either DMSO (1%) or 10 g/ml of nifurtimox (Sigma-Aldrich, St. Louis, MO) were simultaneously performed (Muelas-Serrano et al., 2000). The assay plates were subsequently incubated for 96 h at 27 ◦ C (Pizzolatti et al., 2002; Luize et al., 2005), and the activity of each extract was categorized as low, moderate or high. The inhibitory effect on cell growth was estimated via the colorimetric tetrazolium dye (MTT) method (Muelas-Serrano et al., 2000; Machado et al., 2010). The concentrations inhibiting culture growth were determined through a dose–response regression analysis to obtain IC50 values (the concentration required for 50% inhibition, Santos et al., 2012), as plotted by the Prism 6 program (GraphPad Software, Inc., San Diego, CA). Trypanocidal activity was expressed as the percentage of T. cruzi epimastigotes (±SD) that were lysed (Pizzolatti et al., 2002).
Please cite this article in press as: Molina-Garza, Z.J., et al., Anti-Trypanosoma cruzi activity of 10 medicinal plants used in northeast Mexico. Acta Trop. (2014), http://dx.doi.org/10.1016/j.actatropica.2014.04.006
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Table 1 In vitro trypanocidal activity of methanolic extracts of the selected medicinal plants. Plant species
Part studied
% Growth inhibition ± SD 150 g/ml
Artemisia mexicana Castela texana Cymbopogon citrates Eryngium heterophyllum Haematoxylum brasiletto Lippia graveolens Marrubium vulgare Persea americana Ruta chalepensis Schinus molle Control (Nifurtimox)
AP L AP AP B L AP L L AP
83.93 32.44 83.30 88.38 93.04 33.01 98.99 61.32 75.85 100 80
± ± ± ± ± ± ± ± ± ± ±
0.03 4.06 0.70 0.11 6.9E−4 0.31 0.13 0.45 0.28 0.0 0.0
IC50 (g/ml) (95% CI) 39.25 (8.56–77.07) >150 68.25 (38.36–68.25) 11.24 (6.78–17.40) 7.92 (5.51–10.72) >150 22.66 (17.72–27.06) 65.51 (26.43–294.50) 72.30 (52.38–96.81) 16.31 (12.98–20.35) 10
AP, aerial part; L, leaves; B, bark; CI, confidence intervals.
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3. Results and discussion The in vitro trypanocidal activities of 10 medicinal plants used in northeast Mexico were evaluated. The activities of their methanolic extracts regarding inhibiting the growth of T. cruzi epimastigotes are shown in Table 1. The methanolic extracts of E. heterophyllum, H. brasiletto, M. vulgare and S. molle exhibited the highest activity against T. cruzi, achieving levels of growth inhibition between 88 and 100%. C. citratus and A. mexicana also exhibited good inhibitory activity, with approximately 83% inhibition of epimastigote growth observed under the analyzed concentrations. The P. americana and R. chalepensis extracts displayed a moderate level of activity against T. cruzi epimastigotes, resulting in approximately 70% inhibition. Finally, the methanolic extracts of C. texana and L. graveolens exhibited low activity against the protozoans (ca. 33% growth inhibition). Similar to previous reports, the positive control (nifurtimox) caused 80% growth inhibition at a concentration of 10 g/ml (MuelasSerrano et al., 2000). The methanolic extracts of H. brasiletto and E. heterophyllum showed the highest inhibitory effects against T cruzi at the lowest concentration (IC50 = 7.92 and 11.24 g/ml, respectively). Previous studies have shown that the crude methanolic extract of H. brasiletto can inhibit 100% of T. cruzi growth at 2 mg/ml and 80 to 90% of T. cruzi growth at 1 mg/ml (Abe et al., 2002). For E. heterophyllum, there are no previous studies of trypanocidal activity; however, some species of Eryngium, such as E. carlinae, have previously been reported to present trypanocidal activity (Abe et al., 2005). Additionally, crude extracts of E. amoriginum and E. ternatum have been tested for activity against Leishmania donovani and have shown inhibitory effects, displaying IC50 values between 2.0 and 9.5 g/ml, respectively (Fokialakis et al., 2007). S. molle also demonstrated good activity against T. cruzi (IC50 = 16.31 g/ml) in contrast to the DL50 of 50 g/ml in trypomastigote stage reported by Pizzolatti et al. (2002). This difference may be caused by different concentrations of the bioactive compounds in plants from distinct geographical regions. Otherwise, the essential oil of S. molle possesses trypanocidal activity over T. evansi (Baldissera et al., 2013), T. cruzi, T. brucei, and L. infantum, with an IC50 between 2 and 4.4 g/ml (Abdel-Sattar et al., 2010). In addition, leaf and fruit extracts of S. molle show repellent and insecticidal properties against nymphs and eggs of Triatoma infestans, which are vectors of Chagas disease (Ferrero et al., 2006). The strong inhibitory effects of M. vulgare (presenting IC50 = 22.66 g/ml) on the growth of epimastigotes in this study were of particular interest. In previous reports, the hexane fraction of M. vulgare showed stronger activity than the methanol fraction against T. cruzi epimastigotes (Gohari et al., 2008) and trypomastigotes in vitro (Abe et al., 2005). In addition, Abdel-Sattar et al. (2010) reported high inhibitory effects on the growth of T. brucei (IC50 = 0.25 g/ml) and L. infantum (IC50 = 2.03 g/ml).
We observed an acceptable level of activity in the extract of A. mexicana, which presented an IC50 = 39.25 g/ml. Other studies have shown that A. ludoviciana ssp. mexicana, which is a botanical synonym of A. mexicana, had a growth inhibitory effects of 80–90% at 2 mg/ml on the epimastigote stage of T. cruzi (Abe et al., 2002), which is less active than our extract. In addition, A. indica exhibits an excellent anti-leishmanicidal activity in all Leishmania strains studied, with an IC50 of 0.21 to 0.58 g/ml (Ganguly et al., 2006); and A. vulgari showed a high inhibition level of promastigote culture forms (Kheiri-Manjili et al., 2012). In the present study, the methanolic extract of C. citratus inhibited the growth of epimastigote forms of T. cruzi, presenting an IC50 = 68.25 g/ml, which is slightly higher than the IC50 (63.09 g/ml) observed for the essential oil of C. citratus against epimastigote forms of T. cruzi (Rojas et al., 2010). The same results were obtained with hydroalcoholic extracts of C. citratus, which confirmed that Leishmania species appear to be more sensitive to these extracts than T. cruzi (Luize et al., 2005; Machado et al., 2010). The essential oil of C. citratus also effectively inhibits the growth of L. chagasi (Oliveira et al., 2009). P. americana and R. chalepensis showed growth inhibition at concentrations of 65.51 and 72.30 g/ml (IC50 ), respectively. Moreover, similar results have previously demonstrated the moderate activity of methanolic extracts of the seeds of P. americana against epimastigotes (Abe et al., 2005). For R. chalepensis, there are no previous studies of trypanocidal activity; however, the onset of molting is delayed in Rhodnius prolixus, which is a vector of Chagas disease, when exposed to the odor of R. chalepensis (Abramson et al., 2007). In the present study, the extracts of C. texana and L. graveolens were inactive against T. cruzi epimastigotes. In similar studies, C. texana also showed no activity against epimastigotes (Abe et al., 2005), and hydroalcoholic extracts of L. alba were found to be inactive against L. amazonensis and T. cruzi (Luize et al., 2005). There are reports concerning the role of some plant chemical compositions in the trypanocidal activity of pure compounds isolated from some of the studied species of plants in this current study, including hematoxylin, brazilin, caffeic acid, methyl gallate, gallic acid, phloroglucinol, 4-hydroxycinnamic acid and 5-methoxypsoralen, which were isolated from H. brasiletto (RiveroCruz, 2008). Commercially acquired gallic acid, with an IC50 of 67.0 g/ml (Tasdemir et al., 2006), and 4-hydroxycinnamic acid derivatives isolated from Brazilian propolis (Salomão et al., 2008), showed trypanocidal activity against T. cruzi. Neither chemical nor biological reports concerning the trypanocidal activity of E. heterophyllum could be found in the literature; however, (E)-2-dodecenal isolated from E. foetidum possesses important trypanocidal activity (Forbes and Steglich, 2007). Phytochemical studies of S. molle showed the presence of ␣phellandrene, -phellandrene, ␣-terpineol, ␣-pinene, -pinene and p-cymene (Bendaoud et al., 2010), as well as -caryophyllene, ␣-copaene and ␣-humulene (Díaz et al., 2008; Deveci et al., 2010).
Please cite this article in press as: Molina-Garza, Z.J., et al., Anti-Trypanosoma cruzi activity of 10 medicinal plants used in northeast Mexico. Acta Trop. (2014), http://dx.doi.org/10.1016/j.actatropica.2014.04.006
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The essential oils of Annona pickelii and Annona salzmannii, which contain -caryophyllene, ␣-copaene and ␣-humulene, showed an IC50 lower than 100 g/ml against the trypomastigote stage of T. cruzi (Costa et al., 2013). In a similar manner, caryophylene derivatives present in the essential oil of Hedychium coronarium showed trypanocidal activity against T. brucei (Costa et al., 2013; Rodrigues et al., 2013). From M. vulgare, numerous terpenes, such as ␥-eudesmol, citronellol, citronellyl formate and germacrene D (Kadril et al., 2011), and some flavonoids, such as apigenin, luteolin and vitexin, were isolated (Nawwar et al., 1989). Commercially acquired flavonoids and derivatives, which were found in M. vulgare, possess important trypanocidal activity. This situation is the case for luteolin, with an IC50 of 21.4 g/ml, for luteolin-7-O-glucoside, vitexin and apigenin-7-O-glucoside, with a major IC50 of 30 g/ml, and, finally, apigenin, with an IC50 = 21.8 g/ml (Tasdemir et al., 2006). From A. mexicana, caffeic acid, gallic acid, kampferol, quercetin, myricetin and -caryophyllene (Lopes-Lutz et al., 2008; Carvalho et al., 2011) have been isolated, and there are reports that the commercially acquired compounds possess important trypanocidal activity (Tasdemir et al., 2006; Rodrigues et al., 2013). Citral, which is a major compound found in C. citratus, possesses inhibitory effects in vitro over T. cruzi at a concentration of 40 g/ml at 24 h after application during the differentiation process (Cardoso and Soares, 2010) and kills 65% of L. infantum and L. major and 80% of L. tropica promastigotes at a concentration of 50 g/ml (Machado et al., 2010, 2012). 1,2,4-Trihydroxyheptadecane and 1,2,4-trihydroxynonadecane derivatives obtained from seeds of P. americana exhibit moderate activity against epimastigotes and trypomastigotes of T. cruzi (Abe et al., 2005). Several furanocoumarines and quinolone alkaloids were isolated from R. chalepensis (El-Sayed et al., 2000; Günaydin and Savci, 2005). Some commercially acquired furanocoumarins, such as umbelliferone and bergapten exhibit moderate activity against epimastigotes and trypomastigotes of T. cruzi (Tasdemir et al., 2006). In recent years, several studies have used medicinal plants for the treatment of diseases related to infections with protozoa and with other microorganisms. Cytotoxic activities of crude extracts of various plant species have been demonstrated against amastigotes, epimastigotes and trypomastigotes of T. cruzi (Mafezoli et al., 2000; Pizzolatti et al., 2002; Santoro et al., 2007). Additionally, although the reproducibility of the growth inhibition of T. cruzi culture forms in vitro and in vivo has been previously demonstrated (Schlemper et al., 1977; Abe et al., 2002), some studies have shown that plant extracts exhibit higher trypanocidal activity against some T. cruzi stages in vitro or in vivo (Abe et al., 2005; Castro et al., 1992; Dantas et al., 2006). We consider that advantages of the screening method with T. cruzi epimastigotes include establishing fast and reliable anti-T. cruzi screening programs at low costs, despite the lack of large animal facilities, and the availability of low amounts of drugs or crude extracts. In addition, the manipulation of culture forms is most likely less risky than the handling of infected animals (Schlemper et al., 1977). Several natural products, such as alkaloids, terpenes and quinones, flavonoids and saponins, with trypanocidal activity have been isolated (Wright and Phillipson, 1990; Kayser et al., 2003). Considering these findings, we can conclude that the results obtained in this study indicate that the most promising extracts may result in potential sources of lead compounds for the development of more effective drugs for the treatment of Chagas disease. The bioguided fractionation of E. heterophyllum, H. brasiletto, M. vulgare and S. molle active extracts is ongoing, and further studies of the isolated pure compounds in vivo with amastigotes and trypomastigotes stages of T. cruzi will be performed. Furthermore, these
results suggest that a variety of potentially bioactive substances exist in the Mexican flora.
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We gratefully acknowledge the contributions of QBP Paola Katyana Vázquez-Luévanos and QBP Diana Janeth Coronado˜ who performed the laboratory work. In addition, special Zermeno, thanks go to Dr. José Luis Rosales Encina and Biol. Lidia Baylon (Departamento de Infectómica y Patogénesis Molecular, Centro de Investigación y de Estudios Avanzados, México), who kindly provided us with the CL Brener strain. This project was partially supported by PAICYT-UANL CN835-11. Q2 Q3
References Abdel-Sattar, E., Maes, L., Salama, M.M., 2010. In vitro activities of plant extracts from Saudi Arabia against malaria, leishmaniasis, sleeping sickness and Chagas disease. Phytother. Res. 24, 1322–1328. Adams, M., Chammartin, M., Hamburger, M., Potterat, O., 2013. Case study of the Swiss flora for prior phytochemical and biological investigations. J. Nat. Prod. 76, 209–215. Abe, F., Nagafuji, S., Yamauchi, T., Okabe, H., Maki, J., Higo, H., Akahane, H., Aguilar, A., Jiménez-Estrada, M., Reyes-Chilpa, R., 2002. Trypanocidal constituents in plants. 1. Evaluation of some Mexican plants for their trypanocidal activity and active constituents in Guaco, roots of Aristolochia taliscana. Biol. Pharm. Bull. 25, 1118–1191. Abe, F., Nagafuji, S., Okawa, M., Kinjo, J., Akahane, H., Ogura, T., Martinez-Alfaro, M.A., Reyes-Chilpa, R., 2005. Trypanocidal constituents in plants. 5. Evaluation of some Mexican plants their trypanocidal activity and active constituents in the seeds of Persea americana. Biol. Pharm. Bull. 28, 1314–1317. Abramson, C.H.I., Aldana, E., Sulbaran, E., 2007. Exposure to citral, cinnamon and ruda disrupts the life cycle of a vector of Chagas disease. Am. J. Environ. Sci. 3, 7–8. Alonso-Castro, A.J., Villarreal, M.L., Salazar-Olivo, L.A., Gomez-Sanchez, M., Dominguez, F., Garcia-Carranca, A., 2011. Mexican medicinal plants used for cancer treatment: pharmacological, phytochemical and ethnobotanical studies. J. Ethnopharmacol. 133, 945–972. Baldissera, M.D., Da Silva, A.S., Oliveira, C.B., Zimmermann, C.E.P., Vaucher, R.A., Santos, R.C.V., Rech, V.C., Tonin, A.A., Giongo, J.L., Mattos, C.B., Koester, L., Santuario, J.M., Monteiro, S.G., 2013. Trypanocidal activity of the essential oils in their conventional and nanoemulsion forms: in vitro tests. Exp. Parasitol. 134, 356–361. Bardai, S.E., Lyoussi, B., Wibo, M., Morel, N., 2001. Pharmacological evidence of hypotensive activity of Marrubium vulgare and Foeniculum vulgare in spontaneously hypertensive rat. Clin. Exp. Hypertens. 23, 329–343. Bastos, C.J.C., Aras, R., Mota, G., Reis, F., Pereira Dias, J., Silva de Jesus, R., Silva Freire, M., de Araújo, E.G., Prazeres, J., Rios Grassi, M.F., 2010. Clinical outcomes of thirteen patients with acute Chagas disease acquired through oral transmission from two urban outbreaks in northeastern Brazil. PLoS Negl.Trop. Dis. 4, e711. BDMTM (Biblioteca Digital de la Medicina Tradicional Mexicana), 2013. Eryngium heterophyllum. BDMTM, . Bendaoud, H., Romdhone, M., Souchard, J.P., Cazaux, S., Bouajila, J., 2010. Chemical composition and anticancer and antioxidant activities of Schinus molle L. and Schinus terebinthifolius raddi berries essential oil. J. Food Sci. 75, 466–472. Bern, C., Kjos, S., Yabsley, M.J., Montgomery, S.P., 2011. Trypanosoma cruzi and Chagas’ disease in the United States. Clin. Microbiol. Rev. 24, 655–681. Calzado-Flores, C., Hurtado Ramírez, M.B., Flores Villanueva, Z., Verde Star, M.J., Segura Luna, J.J., 2002. Preliminary morphological study of the effects produced by a methanolic extract of Castela texana. Proc. West. Pharmacol. Soc. 45, 161. Camou-Guerrero, A., Reyes-García, V., Martínez-Ramos, M., Casas, A., 2008. Knowledge and use value of plant species in a Rarámuri community: a gender perspective for conservation. Hum. Ecol. 36, 259–272. Carabarin-Lima, A., González-Vázquez, M.C., Rodríguez-Moralesa, O., BaylónPacheco, L., Rosales-Encina, J.L., Reyes-López, P.A., Arce-Fonseca, M., 2013. Chagas disease (American trypanosomiasis) in Mexico: an update. Acta Trop. 127, 126–135. Cardenas-Sánchez, J.D., Mazariego-Arana, M.A., Utrilla-Pascacio, F.J., MonteónPadilla, V.M., Altúzar-González, M., 2003. Anticuerpos anti-Trypanosoma cruzi en pacientes con cardiomiopatía dilatada. Rev. Med. Mex. 41, 111–114. Cardoso, J., Soares, M.J., 2010. In vitro effects of citral on Trypanosoma cruzi metacyclogenesis. Mem. Inst. Oswaldo Cruz 105, 1026–1032. Carod-Artal, F.J., Vargas, A.P., Horan, T.A., Nadal-Nunes, L.G., 2005. Chagasic cardiomyopathy is independently associated with ischemic stroke in Chagas disease. Stroke 36, 965–970. Carvalho, I.S., Cavaco, T., Brodelius, M., 2011. Phenolic composition and antioxidant capacity of six artemisia species. Ind. Crops Prod. 33, 382–388. Castro, S.L., Soeiro, M.N.C., Meirelles, M.N.L., 1992. Trypanosoma cruzi: effect of phenothiazines on the parasite and its interactions with host cells. Mem. Inst. Oswaldo Cruz 87, 209–215.
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Chagas, C., 1909. Nova tripanozomiase humana: estudos sobre a morfolojia e o ciclo evolutivo do Schizotrypanum cruzi n. gen., n. sp., ajente etiolojico de nova entidade mórbida do homem. Mem. Inst. Oswaldo Cruz 1, 159–218. Costa, E.U., Dutra, L.M., Salvador, M.J., Gunzaga, R.L.H., Gadelha, R.F., Carvalho, J.E., 2013. Chemical composition of the essential oils of Annona pickelii and Annona salzmannii (Annonaceae) and their antitumour and trypanocidal activities. Nat. Prod. Res. 27, 997–1001. Cruz-Reyes, A., Pickering-Lopez, J.M., 2006. Chagas disease in Mexico: an analysis of geographical distribution during the past 76 years—a review. Mem. Inst. Oswaldo Cruz 101, 345. Dang, Q.L., Kwon, H.R., Choi, Y.H., Choi, G.J., Jang, K.S., Park, M.S., Lim, C.H., Ngoc, L.H., Kim, J.C., 2010. Nematicidal activity against Bursaphelenchus xylophilus of isoobtusilactone A isolated from Persea americana. Nematology 12, 247–253. Dantas, A.P., Salomão, K., Barbosa, H.S., De Castro, S.L., 2006. The effect of Bulgarian propolis against Trypanosoma cruzi and during its interaction with host cells. Mem. Inst. Oswaldo Cruz 101, 207–211. Deveci, O., Sukan, A., Tuzun, N., Kocabas, E.E.H., 2010. Chemical composition, repellent and antimicrobial activity of Schinus molle L. J. Med. Plants Res. 4, 2211–2216. Díaz, C., Quesada, S., Brenes, O., Aguilar, G., Cicció, F.J., 2008. Chemical composition of Schinus molle essential oil and its citotoxic activity on tumour cell lines. Nat. Prod. Res. 22, 1521–1534. El-Sayed, K., Al-Said, M.S., El-Feraly, F.S., Ross, S.A., 2000. New quinoline alkaloids from Ruta chalepensis. J. Nat. Prod. 63, 995–997. Efferth, T., 2010. Cancer therapy with natural products and medicinal plants. Planta Med. 76, 1035–1036. Ferrero, A.A., Werdin-González, J.O., Sánchez-Chopa, C., 2006. Biological activity of Schinus molle on Triatoma infestans. Fitoterapia 77, 381–383. Fokialakis, N., Kalpoutzakis, E., Tekwani, B.L., Khan, S.I., Kobaisy, M., Skaltsounis, A.L., Duke, S.O., 2007. Evaluation of the antimalarial and antileishmanial activity of plants from the Greek island of Crete. J. Nat. Med. 61, 38–45. Forbes, W.M., Steglich, C., 2007. Methods of treating infectious diseases. In: United States Patent Application 20090047342. Slippery Rock University (Assignee), Slippery Rock, PA) (filed 08/15). Galaviz-Silva, L., Molina-Garza, D.P., González-Santos, M.A., Mercado-Hernández, R., González-Galaviz, J.R., Rosales-Encina, J.L., Molina-Garza, Z.J., 2009. Update on seroprevalence of anti-Trypanosoma cruzi antibodies among blood donors in northeast Mexico. Am. J. Trop. Med. Hyg. 81, 404–406. Ganguly, S., Bandyopadhyay, S., Bera, A., Chatterjee, M., 2006. Antipromastigote activity of an ethanolic extract of leaves Artemisa indica. Indian J. Pharmacol. 38, 64–65. Gohari, A.R., Saeidnia, S., Hadjiakhoondi, A., Naghinejad, A., Yagura, T., 2008. Trypanocidal activity of some medicinal plants against the epimastigotes of Trypanosoma cruzi. J. Med. Plants 7, 44–48. Günaydin, K., Savci, S., 2005. Phytochemical studies on Ruta chalepensis (Lam.) Lamark. Nat. Prod. Res. 19, 203–210. Gürtler, R.E., Segura, E.L., Cohen, J.E., 2003. Congenital transmission of Trypanosoma cruzi infection in Argentina. Emerg. Infect. Dis. 9, 29–32. Kadril, A., Zarai, Z., Békir, A., Gharsallah, N., Damak, M., Gdoura, R., 2011. Chemical composition and antioxidant activity of Marrubium vugare L. essential oil from Tunisia. Afr. J. Biotech. 10, 3908–3914. Kayser, O., Kiderlen, A.F., Croft, S.I., 2003. Natural products as antiparasitic drugs. Parasitol. Res. 90, S55–S62. Kheiri-Manjili, H., Jafari, H., Ramazani, A., Davoudi, N., 2012. Anti-leishmanial and toxicity activities of some selected Iranian medicinal plants. Parasitol. Res. 111, 2115–2121. Kirchhoff, L.V., Paredes, P., Lomelí-Guerrero, A., Paredes-Espinoza, M., Ron-Guerrero, ˜ ˜ noz, J.G., 2006. Transfusion-associated ChaC.S., Delgado-Mejía, M., Pena-Mu gas disease (American tripanosomiasis) in Mexico: implications for transfusion medicine in the United States. Transfusion 46, 298–304. Lopes-Lutz, D., Alviano, D.S., Alviano, C.S., Kolodziejczyk, P.P., 2008. Screening of chemical composition, antimicrobial and antioxidant activities of artemisia essential oils. Phytochemistry 69, 1732–1738. Luize, P.S., Shioji-Tiuman, T., Morello, L.G., Korehiza Maza, P., 2005. Effects of medicinal plant extracts on growth of Leishmania (L.) amazonensis and Trypanosoma cruzi. Braz. J. Pharm. Sci. 41, 85–94. Machado, M., Santoro, G., Sousa, M.C., Salgueiro, L., Cavaleiro, C., 2010. Activity of essential oils on the growth of Leishmania infantum promastigotes. Flavour Fragance J. 25, 156–160. Machado, M., Pires, P., Dinis, A.M., Santos-Rosa, M., Alves, V., Salgueiro, L., Cavaleiro, C., Sousa, M.C., 2012. Aldheides monoterpenes as potential anti-Leishmania agents: activity of Cymbopongon citratus and Citral on L. infantum, L. tropica and L. major. Exp. Parasitol. 130, 223–231. Mafezoli, J., Vieira, P.C., Fernandes, J.B., da Silva, M.F.G.F., de Albuquerque, S., 2000. In vitro activity of Rutaceae species against the trypomastigote form of Trypanonosma cruzi. J. Ethnopharmacol. 73, 335–340. Martínez-Ibarra, A., Galaviz-Silva, L., Trujilla, C., 1992. Distribución de los triatominos asociados al domicilio humano en el municipio de General Terán, Nuevo León, México. Southwest. Entomol. 17, 261–265. Molina-Garza, Z.J., Galaviz-Silva, L., Molina-Garza, D., Rosales-Encina, J.L., 2007. Prevalencia de Trypanosoma cruzi en triatominos de Nuevo León, México. Salud Pública Mex. 49, 37–44. Monzote, L., Alarcón, O., Setzer, W.N., 2012. Antiprotozoal activity of essential oils review. Agric. Conspec. Sci. 77, 167–175.
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Muelas-Serrano, S., Nogal-Ruiz, J., Gómez-Barrio, A., 2000. Setting of a colorimetric method to determine the viability of Trypanosoma cruzi epimastigotes. Parasitol. Res. 86, 999–1002. Navarro, V., Villarreal, M.L., Rojas, G., Lozoya, X., 1996. Antimicrobial evaluation of some plants used in Mexican traditional medicine for the treatment of infectious diseases. J. Ethnopharmacol. 53, 143–147. Nawwar, M.A.M., El-monsallamy, A.M.D., Barakat, H.H., Buddrus, J., Linscheid, M., 1989. Flavonoid lactates from leaves of Marrubium vulgare. Phytochemistry 28, 3201–3206. Newman, D.J., Cragg, G.M., 2012. Natural products as sources of new drugs over the 30 years from 1981 to 2010. J. Org. Chem. 75, 311–335. PAHO (Pan American Health Organization), 2013. Infographic: Chagas in the Americas. Factsheet-chagas-eng-2013. PAHO, . Pérez-Castorena, A.L., Oropeza, R.F., Vázquez, A.R., Martínez, M., Maldonado, E., 2006. Labdanes and withanolides from physalis coztomatl. J. Nat. Prod. 69, 1029–1033. Pizzolatti, M.G., Koga, A.H., Grisard, E.C., Steindel, M., 2002. Trypanocidal activity of extracts from Brazilian Atlantic rain forest plant species. Phytomedicine 9, 422–426. Quintanilla-Licea, R., Morado-Castillo, R., Gomez-Flores, R., Laatsch, H., Verde-Star, M.J., Hernández-Martínez, H., Tamez-Guerra, P., Tamez-Guerra, R., RodríguezPadilla, C., 2012. Bioassay-guided isolation and identification of cytotoxic compounds from Gymnosperma glutinosum leaves. Molecules 17, 11229–11241. Oliveira, V.C.S., Moura, D.M.S., Lopes, J.A.D., de Andrade, P.P., da Silva, N.H., Figueiredo, R.C.B.Q., 2009. Effects of essential oils from Cymbopogon citratus (DC) Stapf., Lippia sidoides Cham., and Ocimum gratissimum L. on growth and ultrastructure of Leishmania chagasi promastigotes. Parasitol. Res. 104, 1053–1059. Reisenman, C.E., Lawrence, G., Guerenstein, P.G., Gregory, T., Dotson, E., Hildebrand, J.G., 2010. Infection of kissing bugs with Trypanosoma cruzi, Tucson Arizona, USA. Emerg. Infect. Dis. 16, 400–405. Rivero-Cruz, J.F., 2008. Antimicrobial compounds isolated from Haematoxylon brasiletto. J. Ethnopharmacol. 119, 99–103. Rodrigues, D.F., Arenas, V.A.M., Cavaleiro, C., Salgueiro, L., Zimmermann, M.G., Oliveira, M.N., Guncalves, M.M.B., Barretto, C.R.M., Duarte, M.R.R., 2013. Chemical composition and trypanocidal activity of the essential oils from Hedychium coronarium J. Koening (Zingiberaceae). ISRN Infect. Dis. 2013, 1–6. Rojas, J., Solís, H., Palacios, O., 2010. Evaluación in vitro de la actividad anti Trypanosoma cruzi de aceites esenciales de diez plantas medicinales. Anal. Facult. Med. 71, 161–165. ˜ Y., Mejía, A., Díaz-Ruiz, G., Aguilar, M.S., Sánchez-Nieto, S., RiveroRosas-Pinón, Cruz, J.F., 2012. Ethnobotanical survey and antibacterial activity of plants used in the Altiplane region of Mexico for the treatment of oral cavity infections. J. Ethnopharmacol. 141, 860–865. Salomão, K., Pereira, S.P.R., Campos, L.C., Borba, C.M., Cabello, P.H., Marcucci, M.C., de Castro, S.L., 2008. Brazilian Propolis: correlation between chemical composition and antimicrobial anctivity. eCAM. , 317–324. Santoro, G.F., Cardoso, M.G., Guimarães, L.G., Freire, J.M., Soares, M.J., 2007. Anti-proliferative effect of essential oil of Cymbopogon citratus (DC) Stapf (Lemongrass) on intracellular amastigotes, bloodstream trypomastigotes and culture epimastigotes of Trypanosoma cruzi (Protozoa:Kinetoplastida). Parasitology 134, 1649–1656. Santos, K,K,A., Matias, E.F.F., Tintino, S.R., Souza, C.E.S., Braga, M.F.B., Guedes, G.M.M., Rolón, M., Vega, C., de Arias, A.R., Costa, J.G.M., Menezes, I.R.A., Coutinho, H.D.M., 2012. Anti-Trypanosoma cruzi and cytotoxic activities of Eugenia uniflora L. Exp. Parasitol. 131, 130–132. Schlemper, B.R., Chiari, E., Brener, Z., 1977. Growth-inhibition drug test with Trypanosoma cruzi culture forms. J. Protozool. 24, 544–547. Schmunis, G.A., 2007. Epidemiology of Chagas disease in non-endemic countries: the role of international migration. Mem. Inst. Oswaldo Cruz. 102, 75–85. Sülsen, V., Güida, C., Coussio, J., Paveto, C., Muschietti, L., Martino, V., 2006. In vitro evaluation of trypanocidal activity in plants used in Argentine traditional medicine. Parasitol. Res. 98, 370–374. Tasdemir, D., Kaiser, M., Brun, R., Yardley, V., Schmidt, T.J., Tosun, F., Rüedi, P., 2006. Antitrypanosomal and antileishmanial activities of flavonoids and their analogues: in vitro, in vivo, structure activity relationship studies. Antimicrob. Agents Chemother. 50, 1352–1364. Troullier, P., Olliaro, P., Torreele, E., Orbinski, J., Laing, R., Ford, N., 2002. Drug development for neglected diseases: a deficient market and a public health policy failure. Lancet 359, 2188–2194. ˜ D.L., Robledo, S.M., Echeverri, F., Arango, G.J., Vélez, I.D., Triana, Valencia, L., Munoz, O., 2011. Actividad tripanocida y citotóxica de extractos de plantas colombianas. Biomédica 31, 552–559. Wang, X., Beno-Movalem, D., Kobiler, I., Leikin-Frenkel, A., Lichter, A., Prusky, D., 2004. Expression of 12 fatty acid desaturase during the induced accumulation of the antifungal diene in avocado fruits. Mol. Plant Pathol. 5, 575–585. WHO (World Health Organization), 2014. Chagas disease (American trypanosomiasis). In: Fact Sheet No 340, , (updated March, 2013). Wright, C.W., Phillipson, J.D., 1990. Natural products and the development of selective antiprotozal drugs. Phytother. Res. 4, 127–139. ˜ S., Giner, R.M., Cerdá-Nicolás, M., Ríos, J.L., 2003. IsoYueqin, Z., Recio, M.C., Mánez, lation of two triterpenoids and a bioflavanone with anti-inflammatory activity from Schinus molle fruits. Planta Med. 69, 893–898.
Please cite this article in press as: Molina-Garza, Z.J., et al., Anti-Trypanosoma cruzi activity of 10 medicinal plants used in northeast Mexico. Acta Trop. (2014), http://dx.doi.org/10.1016/j.actatropica.2014.04.006
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