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Argemone Mexicana on Trophozoites of Entamoeba Histolytica HM1-IMSS

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Argemone Mexicana on Trophozoites of Entamoeba Histolytica HM1-IMSS In vitro study of antiamoebic activity of methanol extracts of Argemone mexicana on trophozoites of Entamoeba histolytica HM1-IMSS Joel H. Elizondo-Luevano, 1 Magda E. Hernández-García, 2 Rocío Castro-Ríos, 1 Azucena del C. González-Horta, 1 Eduardo Sánchez García, 1Abelardo Chavez-Montes,1 1. Facultad de Ciencias Biológicas, UANL. Av. Pedro de Alba S/N, Cd. Universitaria, 66455 San Nicolás de los Garza. 2. Laboratorio de Biología Celular del Centro de Investigación Biomédica del Noreste, IMSS. Nuevo León, México Institution. Email: [email protected] , [email protected] Abstract Amoebiasis, a parasitic infection caused by Entamoeba histolytica, is considered endemic in México, where Argemone mexicana has been used in traditional medicine to treat intestinal parasitic diseases. The objective of this work was to evaluate the potential biological activity of A. mexicana on E. histolytica. For this purpose, a methanolic extract was prepared from A. mexicana leaves, and a differential fractionation was carried out with solvents of different polarities. The inhibitory capacities of the extract were evaluated in vitro using HM1-IMSS, a strain of Entamoeba histolytica. A. mexicana extract was found to have a growth-inhibiting activity for E. histolytica, showing an IC50 = 78.39 μg/mL. The extract was characterized phytochemically and the methanolic extract fractions were analyzed by liquid chromatography (HPLC) and mass spectrometry (MS). Berberine and jatrorrhizine were present in the active fractions, and these compounds may be responsible for the antiparasitic activity. The identification of amoebicidal activity of A. mexicana on E. histolytica gives support to the traditional use. Introduction Statistical analysis E. histolytica is one of the most common parasites and is the causative agent of human amoebiasis [1]. Approximately 50 million people are affected by E. histolytica each year, and of these, approximately 100,000 die [2]. In fact, intestinal infections by protozoa, such as amoebiasis, are the third leading cause of death in the world [3]. Materials and Methods Leaves of A. mexicana were collected in the city of Monterrey, Nuevo HPLC analysis and mass spectrometry. León, Mexico, in February 2017. Argemone mexicana Scientific classification Kingdom Plantae Clade Angiosperms Clade Eudicots Order Ranunculales Family Papaveraceae Genus Argemone Species A. mexicana Binomial name Argemone mexicana L. Nuclear Magnetic Resonance Spectroscopy (NMR) Extraction of Dried plant ( Leaves ) solvent • A.mexicana • TLC (Thin-layer chromatography) Fractions • Phytochemical tests •Hexane z •Chlorophorm •MeOH Maceration •Hot MeOH •Aqueous MeOH Thin-layer chromatography of the methanolic extract of A. mexicana Chemical shift values of 1H and 13C NMR for berberine 1. 2. 3. 1. 2. 3. 1. MeOH EXTRACT 2. BERBERINE 3. JATRORRHIZINE Assignment of hydrogen atoms Assignment of carbon atoms z Conclusions: The methanolic extract of A. mexicana showed growth inhibition activity against trophozoites of the E. histolytica strain HM1:IMSS. The analysis of the fractions by HPLC-MS showed the presence of berberine alkaloids and jatrorrhizine. References Financial support and acknowledgments [1] Pineda, E, and Doranda P. 2017. “Entamoeba Histolytica under Oxidative Stress: What We thank the financial support of the National Council of Science and Countermeasure Mechanisms Are in Place?” Cells 6 (4): 44. [2] FAO/WHO. 2014. Multicriteria-Based Ranking for Risk Management of Food-Borne Parasites. Technology (CONACYT) through project CB176853 and the subsidy received http://www.fao.org/3/a-i3649e.pdf%5Cnfiles/540/a-i3649e.pdf. for Joel H. Elizondo Luevano. Authors thank M.C. Sergio García and Dr. [3] B. Mata-Cárdenas, J. Vargas-Villarreal, F. González-Salazar and S. Said-Fernández. 2008. “A Roberto Mercado for their invaluable technical assistance. New Vial Microassay to Screen Antiprotozoal Drugs.”Pharmacologyonline 1: 529 – 37..
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