Journal of Ethnopharmacology 112 (2007) 415–416
Ethnopharmacological communication
Antiprotozoal activity of Senna racemosa Rosa E. Moo-Puc c,∗ , Gonzalo J. Mena-Rejon a , Leovigildo Quijano b , Roberto Cedillo-Rivera c a
b
Facultad de Qu´ımica, Universidad Aut´onoma de Yucat´an, 41 No. 42 Col. Industrial, 97150 M´erida, Yucat´an, Mexico Instituto de Qu´ımica, Universidad Nacional Aut´onoma de M´exico, Circuito Exterior, Ciudad Universitaria, Coyoacan, 04510 Mexico City, Mexico c Unidad Interinstitucional de Investigaci´ on Cl´ınica y Epidemiol´ogica, Facultad de Medicina, UADY/Instituto Mexicano del Seguro Social, M´erida, Yucat´an, Mexico Received 28 October 2006; received in revised form 20 March 2007; accepted 25 March 2007 Available online 30 March 2007
Abstract Methanol extracts of leaves, roots and bark of Senna racemosa (Mill.) H.S. Irwin & Barneby (syn. Cassia racemosa Mill.) were tested for antiprotozooal activity against Giardia intestinalis and Entamoeba histolytica. All of the tested extracts showed good activity against both protozoa species. Extracts from stem bark and leaves were most active, with an IC50 of 2.10 g/mL for Giardia intestinalis and 3.87 g/mL for Entamoeba histolytica. Of the previously isolated compounds from Senna racemosa, the piperidine alkaloid cassine had greater activity against Giardia intestinalis with an IC50 of 3.28 g/mL and chrysophanol, a 1,8-dihydroxy-anthraquinone, was the most active agent against Entamoeba histolytica, with an IC50 of 6.21 g/mL. © 2007 Elsevier Ireland Ltd. All rights reserved. Keywords: Senna racemosa; Fabaceae; Dihydroxyanthraquinones; Cassine; Antigiardial; Antiamoebic
1. Plant material
3. Previously isolated constituents
Senna racemosa (Mill.) H.S. Irwin & Barneby (syn. Cassia racemosa Mill.) (Irwing and Barneby, 1982) was collected in M´erida, Yucat´an, M´exico (21◦ 58 N, 89◦ 36 W) during June 2005. The samples were authenticated by Jos´e Salvador Flores Guido at the Botany Department, Faculty of Veterinary Medicine, Autonomous University of Yucatan (UADY). A voucher specimen (J.S. Flores No. 12,704) was deposited at UADY’s Alfredo Barrera Mar´ın Herbarium.
Previous phytochemical investigations revealed the presence of the piperidine alkaloid cassine, alditol, pinitol, the anthraquinones physcion and chrysophanol, and racemochrysone, a dihydroanthracenone derivative (Mena-Rejon et al., 2002).
2. Uses in traditional medicine Senna racemosa is a Fabaceae species known as K’aanlool, K’aan-lool-che and Ja’abin-peek in their native Yucatan Peninsula, Mexico (Arellano-Rodriguez et al., 2003). These plants have been used in Mayan folk medicine for treatment of several ailments. Leaf, root and bark infusions are recommended for treatment of diarrhea, eye infections, diabetes, earache, headache, molar, abdominal and epigastric pain (Rosado-Vallado et al., 2000; Flores, 2001). ∗
Corresponding author. Tel.: +52 999 924 0554; fax: +52 999 923 3297. E-mail address:
[email protected] (R.E. Moo-Puc).
0378-8741/$ – see front matter © 2007 Elsevier Ireland Ltd. All rights reserved. doi:10.1016/j.jep.2007.03.028
4. Materials and methods 4.1. Preparation of crude extracts and isolation of compounds After drying in the shade, the leaves, bark and roots of Senna racemosa were ground and the powdered plant material was extracted with methanol in a Soxhlet apparatus. The solvent was evaporated in a rotatory evaporator and the residue yield was calculated as 5.15 g, 20.6% (w/w) for leaves, 1.16 g, 5.8% (w/w) for bark and 1.46 g, 7.3% (w/w) for roots. Composition analysis yielded physcion (0.11%, w/w) and chrysophanol (0.30%, w/w) from the bark and cassine (0.23%, w/w) and pinitol (15%, w/w) from the leaves, as previously isolated by Mena-Rejon et al. (2002).
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R.E. Moo-Puc et al. / Journal of Ethnopharmacology 112 (2007) 415–416
4.2. Susceptibility assays All experiments were performed in triplicate. Cultures of the intestinal parasites Giardia intestinalis IMSS:0696:1 and Entamoeba histolytica HM1-IMSS, were used in this study (Cedillo-Rivera et al., 2003). Their trophozoites were grown in TYI-S-33 modified medium, supplemented with 10% calf serum and TYI-S-33 medium, supplement with 10% bovine serum, respectively. The in vitro susceptibility assays were carried out following a method previously described (Andrzejewska et al., 2004). Ten milligrams extracts were dissolved in 2 mL of dimethylsulfoxide (DMSO) and added to microtubes containing 1.5 mL of medium in order to reach concentrations of 1.6, 3.3, 6.6 and 13.3 g/mL. The solutions were inoculated with Giardia intestinalis or Entamoeba histolytica to achieve an inoculum of 5 × 104 and 6 × 103 trophozoites/mL, respectively. Metronidazole was used as the reference drug, culture medium with trophozoites and DMSO was the negative control, and culture medium was the blank. Inoculated solutions were incubated for 48 h at 37 ◦ C. After, parasites were detached by chilling and trophozoites were counted with a haemocytometer.
the case of Entamoeba histolytica the leaves extract was the most active (3.87 g/mL), but also the activity of roots and bark extracts was important (4.86 and 7.15 g/mL, respectively). Among the isolated compounds, cassine showed greater activity against Giardia intestinalis. Since cassine itself is almost as active as the leaves and bark extracts, suggesting that their activity is due in part by the presence of the piperidinic alkaloid. In the case of Entamoeba histolytica, only chrysophanol showed only moderate activity, suggesting that the antiamoebic activity of the extracts is due to other compounds. Although chrysophanol and physcion are structurally similar anthraquinones, the observed antiprotozoal activity reveals that small structural differences such as the presence or absence of a methoxy group could determine the selectivity of the antiprotozoal activity, thus, physcion was more active against Giardia intestinalis, while chrysophanol was more active against E. histolitytica. The results obtained in the present investigation, together with the reported antimicrobial activity of the leaves extracts (Rosado-Vallado et al., 2000) and that of the piperidinic alkaloid cassine (Sansores-Peraza et al., 2000), support the use of this plant for the treatment of diarrhea in traditional Mayan medicine.
4.3. Statistical analysis Acknowledgements The data were analyzed using Probit analysis. The percentage of trophozoite survivors was calculated by comparison with growth in the control group. The IC50 and the 95% confidence limit were computed from a plot of probit against the drug concentration.
We thank the Mexican IMSS-FOFOI and the Consejo Nacional de Ciencia y Tecnolog´ıa (CONACyT) by their financial support of projects 2005/6/1/471 and 0276PB, and Dr. Manuel Flores-Arce for English revision of the manuscript.
5. Results and discussion
References
All tested extracts showed good in vitro activity against Giardia intestinalis and Entamoeba histolytica trophozoites (Table 1). The bark extract was the most active against Giardia intestinalis with a IC50 of 2.10 g/mL. The activity of the leaves and roots, however, was also important, with an IC50 of 3.58 g/mL for leaves and 5.19 g/mL for root extracts. In
Andrzejewska, M., Yepez-Mulia, L., Tapia, A., Cedillo-Rivera, R., Laudy, A.E., Starosciak, B.J., Kazimierczuk, Z., 2004. Synthesis, and antiprotozoal and antibacterial activities of S-substituted 4,6-dibromo- and 4,6-dichloro-2mercaptobenzimidazoles. European Journal of Pharmaceutical Science 21, 323–329. Arellano-Rodriguez, J.A., Flores-Guido, J.S., Tun-Garrido, J., Cruz-Bojorquez, M.M., 2003. Nomenclatura, forma de vida, uso, manejo y distribuci´on de las especies vegetales de la pen´ınsula de Yucat´an. Etnoflora Yucatense. Universidad Aut´onoma de Yucat´an, p. 381. Cedillo-Rivera, R., Darby, J.M., Enciso-Moreno, J.A., Ortega-Pierres, G., 2003. Genetic homogeneity of axenic isolates of Giardia intestinalis derived from acute and chronically infected individuals in Mexico. Parasitology Research 90, 119–123. Flores, J.S., 2001. Leguminosae:Flor´ıstica. Etnobot´anica y Ecolog´ıa. Etnoflora Yucatanense. Universidad Aut´onoma de Yucat´an, p. 291. Irwing, H.S., Barneby, R.C., 1982. The american Cassinae a sinoptical revision of leguminosae tribe Cassiae subtripe Cassinae in the new world. Memoirs of the New York Botanical Garden 35, 454. Mena-Rejon, G.J., Perez-Rivas, K., Sansorez-Peraza, P., Rios, T., Quijano, L., 2002. Racemochrysone, a dihydroanthracenone from Senna racemosa. Zeitschrift f¨ur Naturforschung C 57, 777–779. Rosado-Vallado, M., Brito-Loeza, W., Mena-Rejon, G.J., Quintero-Marmol, E., Flores-Guido, J.S., 2000. Antimicrobial activity of Fabaceae species used in Yucatan traditional medicine. Fitoterapia 71, 570–573. Sansores-Peraza, P., Rosado-Vallado, M., Brito-Loeza, W., Mena-Rejon, G.J., Quijano, L., 2000. Cassine, an antimicrobial alkaloid from Senna racemosa. Fitoterapia 71, 690–692.
Table 1 Antiprotozoal activity of methanol extracts and compounds from Senna racemosa Plant part/compound
Leaves Bark Roots Chrysophanol Physcion Pinitol Cassine Metronidazoleb a b
IC50 g/mL (CI)a Giardia intestinalis
Entamoeba histolytica
3.58 (3.57–3.60) 2.10 (2.09–2.11) 5.19 (5.18–5.20) 12.70 (12.62–12.76) 5.27 (5.25–5.28) 11.70 (11.66–11.72) 3.28 (3.25–3.31) 0.160 (0.159–0.163)
3.87 (3.86–3.89) 7.15 (7.13–7.17) 4.86 (4.83–4.87) 6.21 (6.19–6.22) 12.70 (12.66–12.74) 16.59 (16.46–16.72) 11.96 (11.93–12.00) 0.030 (0.029–0.034)
CI = 95% confidence interval. Positive control.
