Back
 Health  Vol.9 No.7 , July 2017
Antiprotozoal Activity of a Thymus vulgaris Methanol Extract and Its Fractions
Abstract:
Introduction: Thymus vulgaris is used in traditional medicine to treat gastrointestinal diseases because of its antifungal, antibacterial, and antispasmodic activity. Objective: To verify whether Thymus vulgaris also has antiprotozoal activity against Trichomonas vaginalis, Giardia lamblia and Entamoeba histolytica trophozoites. Materials and methods: Conventional cultures of parasites were measured on the third day during the logarithmic growth phase. The antiprotozoal activity of the methanol extract and its fractions were evaluated comparing growth in cultures with and without extracts. Next, the extract was fractionated by polarity-based partitioning. Then, the purity of each fraction was determined by thin layer chromatography (TLC). The percentage of growth inhibition was calculated with respect to untreated controls. The 50% inhibitory concentration (IC50) of each extract was calculated by PROBIT analysis. Results: We found that a methanol extract of the aerial parts of Thymus vulgaris, at 300 μg/mL, inhibited the in vitro growth of G. lamblia and T. vaginalis, while E. histolytica growth was poorly inhibited. The methanol extract was further separated into mixtures of ursolic, oleanolic, and betulinic acids. The IC50 values of ursolic acid against G. lamblia and T. vaginalis were 8.12 μg/mL and 5.51 μg/mL, respectively. Conclusions: The methanol extract fraction containing ursolic acid obtained from Thymus vulgaris has antiprotozoal activity against G. lamblia and T. vaginalis trophozoites.
Cite this paper: Garza-González, J. , Vargas-Villarreal, J. , Verde-Star, M. , Rivas-Morales, C. , Oranday-Cárdenas, A. , Hernandez-García, M. , Garza-Salinas, L. and González-Salazar, F. (2017) Antiprotozoal Activity of a Thymus vulgaris Methanol Extract and Its Fractions. Health, 9, 1081-1094. doi: 10.4236/health.2017.97079.
References

[1]   Hernández-Cortéz, C., Aguilera, A.M.G. and Castro, E.G. (2011) Situación de las enfermedades gastrointestinales en México. Enf Infect and Microb, 31, 137-151.

[2]   Adam, R.D. (2001) Biology of Giardia lamblia. Clinical Microbiology Reviews, 14, 447-475.

[3]   Lane, S. and Lloyd, D. (2002) Current Trends in Research into the Waterborne Parasite Giardia. Critical Reviews in Microbiology, 28, 123-147.
https://doi.org/10.1080/1040-840291046713

[4]   Thompson, R.C. and Monis, P.T. (2004) Variation in Giardia: Implications for Taxonomy and Epidemiology. Advances in Parasitology, 58, 69-137.
https://doi.org/10.1016/S0065-308X(04)58002-8

[5]   (1996) The World Health Report. World Health Organization, Geneva.

[6]   Cedillo-Rivera, R., Leal, Y.A., Yépez-Mulia, L., Gómez-Delgado, A., Ortega-Pierres, G., Tapia-Conyer, R. and Munoz, O. (2009) Seroepidemiology of Giardiasis in Mexico. American Journal of Tropical Medicine and Hygiene, 80, 6-10.

[7]   Chacín-Bonilla, L. (2013) An Update on Amebiasis. Revista Médica de Chile, 41, 609-615.
https://doi.org/10.4067/S0034-98872013000500009

[8]   Petri, W.A.Jr. (1996) Recent Advances in Amebiasis. Critical Reviews in Clinical Laboratory Sciences, 33, 1-37.
https://doi.org/10.3109/10408369609101485

[9]   Caballero-Salcedo, A., Viveros-Rogel, M., Salvatierra, B., Tapia-Conyer, R., Sepúlveda-Amor, J. and Gutiérrez, G. (1994) Sero-Epidemiology of Amebiasis in México. American Journal of Tropical Medicine and Hygiene, 50, 412-419.
https://doi.org/10.4269/ajtmh.1994.50.412

[10]   Stanley, S.L.Jr. (2003) Amoebiasis. The Lancet, 361, 1025-1034.
https://doi.org/10.1016/S0140-6736(03)12830-9

[11]   World Health Organization (2001) Global Prevalence and Incidence of Selected Curable Sexually Transmitted Infections.

[12]   Petri, W.A. (2003) Therapy of Intestinal Protozoa. Trends in Parasitology, 19, 523-526.

[13]   Busatti, H.G., Santos, J.F. and Gomes, M.A. (2009) The Old and New Therapeutic Approaches to the Treatment of Giardiasis: Where Are We? Biologics, 3, 273-287.

[14]   Sobel, J.D., Nyirjesy, P. and Brown, W. (2001) Tinidazole Therapy for Metronidazole-Resistant Vaginal Trichomoniasis. Clinical Infectious Diseases, 33, 1341-1346.
https://doi.org/10.1086/323034

[15]   Lofmark, S., Edlund, C. and Nord, C.E. (2010) Metronidazole Is Still the Drug of Choice for Treatment of Anaerobic Infections. Clinical Infectious Diseases, 50, S16-S23.
https://doi.org/10.1086/647939

[16]   Upcroft, J.A., Campbell, R.W., Benakli, K., Upcroft, P. and Vanelle, P. (1999) The Efficacy of New 5-Nitroimidazoles against Metronidazole-Susceptible and -Resistant Anaerobic Protozoa. Antimicrobial Agents and Chemotherapy, 43, 73-76.

[17]   Canigueral, S. and Vanaclocha, B. (2000) Usos terapéuticos del tomillo. Fitoterapia, 1, 5-13.

[18]   Jiménez-Arellanes, A., Martínez, R., García, R., León-Díaz, R., Luna-Herrea, J., Molina-Salinas, G. and Said-Fernandez, S. (2006) Thymus vulgaris as Potential Source of Antituberculous Compounds. Pharmacologyonline, 3, 569-574.

[19]   Nilforoushzadeh, M.A., Shirani-Bidabadi, L., Zolfaghari-Baghbaderani, A., Saberi, S., Siadat, A.H. and Mahmoudi, M. (2008) Comparison of Thymus vulgaris (Thyme), Achilleamillefolium (Yarrow) and Propolishydroalcoholic Extracts versus Systemic Glucantime in Treatment of Cutaneous leishmaniasis in Balb/C Mice. Journal of Vector Borne Diseases, 45, 301-306.

[20]   Al Maqtari, M.A., Alghalibi, S.M. and Alhamzy, E.H. (2011) Chemical Composition and Antimicrobial Activity of Essential Oil of Thymus vulgaris from Yemen. Turkish Journal of Biochemistry, 36, 342-349.

[21]   Valizadegan, O. (2013) Study on the Influence of Thyme (Thymus vulgaris) Extract on Fungal Control of Some Crop Seeds during Germination Stage. Advances in Environmental Biology, 7, 109-112.

[22]   Said-Fernández, S., Vargas-Villarreal, J., Castro-Garza, J., Mata-Cárdenas, B.D., Navarro-Marmolejo, L., Lozano-Garza, G. and Martínez-Rodríguez, H. (1988) PEHPS Medium: An Alternative for Axenic Cultivation of Entamoeba histolytica and E. invadens. Transactions of the Royal Society of Tropical Medicine and Hygiene, 82, 249-253.

[23]   Molina-Salinas, G.M., Pena-Rodríguez, L.M., Mata-Cárdenas, B.D., Escalante-Erosa, F., González-Hernández, S., Torres de la Cruz, V.M., Martínez-Rodríguez, H.G. and Said-Fernández, S. (2011) Flourensia cernua: Hexane Extracts a Very Active Mycobactericidal Fraction from an Inactive Leaf Decoction against Pan-Sensitive and Pan-Resistant Mycobacterium tuberculosis. Evidence-Based Complementary and Alternative Medicine, 2011, Article ID: 782503.
https://doi.org/10.1155/2011/782503

[24]   Mata-Cárdenas, B.D., Vargas-Villarreal, J., González-Salazar, F., Palacios-Corona, R. and Said-Fernández, S. (2008) A New Vial Microassay to Screen Antiprotozoal Drugs. Pharmacologyonline, 1, 529-537.

[25]   Calzada, F., Yépez-Mulia, L. and Aguilar, A. (2006) In Vitro Susceptibility of Entamoeba histolytica and Giardia lamblia to Plants Used in Mexican Traditional Medicine for the Treatment of Gastrointestinal Disorders. Journal of Ethnopharmacology, 108, 367-370.

[26]   Cedillo-Rivera, R. and Munoz, O. (1992) In Vitro Susceptibility of Giardia lamblia to Albendazole, Mebendazole and Other Chemotherapeutic Agents. Journal of Medical Microbiology, 37, 221-224.
https://doi.org/10.1099/00222615-37-3-221

[27]   Finney, D.J. (1971) Probit Analysis. 3rd Edition, Cambridge University Press, London.

[28]   Reynolds, C.P., Black, A.T. and Woody, J.N. (1986) Sensitive Method for Detecting Viable Cells Seeded into Bone Marrow. Cancer Research, 46, 5878-5881.

[29]   González-Garza, M.T., Mata-Cárdenas, B.D. and Said-Fernández, S. (1989) High Susceptibility of Five Axenic Entamoeba Histolytica Strains to Gossypol. Transactions of the Royal Society of Tropical Medicine and Hygiene, 83, 522-524.

[30]   Oluwatosin, K.S., Habibat, U. and Usman, Y.U. (2013) Effect of Methanolic Leaf Extract of Thymus vulgaris on Some Biomarker Enzymes in Trypanosoma brucei Infected Rats. International Journal of Pharmaceutical and Biomedical Research, 4, 83-87.

[31]   Ríos, J.L. and Recio, M.C. (2005) Medicinal Plants and Antimicrobial Activity. Journal of Ethnopharmacology, 100, 80-84.

[32]   Chun, H., Shin, D.H., Hong, B.S., Cho, H.Y. and Yang, H.C. (2001) Purification and Biological Activity of Acidic Polysaccharide from Leaves of Thymus vulgaris L. Biological and Pharmaceutical Bulletin, 24, 941-946.
https://doi.org/10.1248/bpb.24.941

[33]   Morimitsu, Y., Yoshida, K., Esaki, S. and Hirota, A. (1995) Protein Glycation Inhibitors from Thyme (Thymus vulgaris). Bioscience, Biotechnology, and Biochemistry, 59, 2018-2021.
https://doi.org/10.1271/bbb.59.2018

[34]   Mahboubi, A., Kamalinejad, M., Ayatollahi, A.M. and Babaeian, M. (2014) Total Phenolic Content and Antibacterial Activity of Five Plants of Labiatae against Four Foodborne and Some Other Bacteria. Iranian Journal of Pharmaceutical Research, 13, 559-566.

[35]   Calzada, F., Yépez-Mulia, L. and Tapia-Contreras, A. (2007) Effect of Mexican Medicinal Plant Used to Treat Trichomoniasis on Trichomonas vaginalis Trophozoites. Journal of Ethnopharmacology, 113, 248-251.

[36]   Touchstone, J.C. (1992) Practice of Thin Layer Chromatography. 3rd Edition, John Wiley & Sons, INC., Hoboken.

[37]   Cannell, R.J.P. (1998) Natural Products Isolation. Humana Press Inc., Clifton, 165-208.
https://doi.org/10.1007/978-1-59259-256-2

[38]   Sáenz-Esqueda, M.A., álvarez-Roman, R., Castro-Ríos, R., Gómez-Flores, R., Nunez-Rodríguez, M.A., Galindo-Rodríguez, S.A. and Chávez-Montes, A. (2012) Actividad Antituberculosa del extracto de Carya illinoensis. Revista Mexicana de Ciencias Farmacéuticas, 43, 12-21.

[39]   Machado, M., Dinis, A.M., Salgueiro, L., Cavaleiro, C., Custódio, J.B.A. and Sousa, M.C. (2010) Anti-Giardia Activity of Phenolic-Rich Essential Oils: Effects of Thymbra capitata, Origanum virens, Thymus zigis subsp. Sylvestris and Lippia graveolens on Trophozoites Growth, Viability, Adherence and Ultrastructure. Parasitology Research, 106, 1205-1215.
https://doi.org/10.1007/s00436-010-1800-7

[40]   Al Musayeib, N.M., Mothana, R.A., Gamal, A.A., Al-Massarani, S.M. and Maes, L. (2013) In Vitro Antiprotozoal Activity of Triterpenoid Constituents of Kleinia odora Growing in Saudi Arabia. Molecules, 18, 9207-9218.
https://doi.org/10.3390/molecules18089207

[41]   Behnia, M., Haghighi, A., Komeylizadeh, H., Javad, S., Tabael, S. and Abadi, A. (2008) Inhibitory Effects of Iranian Thymus vulgaris Extracts on in Vitro Growth of Entamoeba histolytica. The Korean Journal of Parasitology, 46, 153-156.
https://doi.org/10.3347/kjp.2008.46.3.153

[42]   Sonboli, A., Mirjalili, M.H., Hadian, J. and Yousefzadi, M. (2014) The Biological Activity and Composition of the Essential Oil of Sclerorhachis leptoclada (Asteraceae-Anthemideae) from Iran. Iranian Journal of Pharmaceutical Research, 13, 1097-1104.

[43]   Alderete, J.F. and Pearlman, E. (1984) Pathogenic Trichomonas vaginalis Cytotoxicity to Cell Culture Monolayers. The British Journal of Venereal Diseases, 60, 99-105.

 
 
Top