Caracterización de la actividad tripanocida y citotóxica de aceites esenciales de lippia alba y sus terpenos bioactivos sobre trypanosoma cruzi

dc.contributorGarcía Sánchez, Liliana Torcoroma
dc.contributorLeal Pinto, Sandra-Milena
dc.creatorMoreno Moreno, Erika Marcela
dc.date.accessioned2018-11-23T16:34:43Z
dc.date.available2018-11-23T16:34:43Z
dc.date.created2018-11-23T16:34:43Z
dc.date.issued2017-06-09
dc.identifierT 86.17 M672c
dc.identifierhttps://repositorio.udes.edu.co/handle/001/637
dc.description.abstractLa Enfermedad de Chagas es una infección causada por el protozoario Trypanosoma cruzi, la cual es caracterizada por un prolongado curso silencioso y complejas manifestaciones clínicas. Actualmente, no existe evidencia contundente de quimioterapéuticas eficaces, lo que hace obligatoria la búsqueda de nuevos agentes tripanocidas más efectivos. Bajo este escenario, fármacos extraídos de plantas resultan prometedores, teniendo en cuenta su baja toxicidad y amplio espectro farmacológico. De acuerdo con esto, en este trabajo se evaluó la actividad tripanocida in vitro de aceites esenciales (AEs) de Lippia alba y de sus terpenos bioactivos (óxido de cariofileno, citral, limoneno y carvona), caracterizándose el mecanismo de muerte celular inducido contra T.cruzi. Los resultados fueron expresados como la concentración inhibitoria o citotóxica 50 (CI50 y CC50, respectivamente), realizando regresión sigmoidal mediante el Software estadístico MsxlfitTM. El fenotipo de muerte fue rastreado por microscopía óptica y de fluorescencia, citometría de flujo y electroforesis de ADN. El análisis estadístico se realizó por ANOVA de Welch (software SPSS 15.0). En las condiciones testadas, los AEs y terpenos de L. alba ejercieron efectos inhibitorios distintos en los tres estadios parasitarios evaluados (p=0.000). Los AEs del quimiotipo Citral exhibieron concentraciones inhibitorias medias menores (Epimastigotes: CI50 13,6±1,5μg/mL; Tripomastigotes: CI50 21,9±1,4 μg/mL; Amastigotes: CI50 74,1±4,4μg/mL) que las alcanzadas con los aceites del quimiotipo Carvona (Epimastigotes: CI50 88,2±3,7μg/mL; Tripomastigotes: CI50 44,9±2,5μg/mL; Amastigotes: CI50 >150μg/mL). El limoneno representó el terpeno de mayor selectividad (Epimastigotes: IS 7,1; Tripomastigotes: IS 32,8; Amastigotes: IS 10,3) y menor toxicidad sobre células Vero (Limoneno: CC50 297,1±2,4μg/mL). Además, el limoneno evidenció una interacción sinérgica in vitro con el Benznidazol (Epimastigotes CIF=0,44±0,13; tripomastigotes CIF=0,42±0,10; amastigotes CIF=0,58±0,13μg/mL) y con el óxido de cariofileno (Epimastigotes: CIF 0,49; Tripomastigotes: CIF 0,45; Amastigotes: CIF 0,71μg/mL). Los análisis celulares sugieren que la muerte celular observada en el T. cruzi podría ser mediada por un mecanismo tipo-apoptótico.
dc.description.abstractChagas Disease is caused by infection of Trypanosoma cruzi parasite. In general, the course of the disease is characterized by a prolonged silent phase and a complex clinical presentation. Currently, there is no compelling evidence of effective chemotherapeutics, making compulsory the research on more effective and selective therapies.In this way, plants offer a promissory reserve of bioactive molecules, take into account their low toxicity and width pharmacologic spectrum. In this work, the in vitro trypanocidal activity of Lippia alba essential oils (EOs) and their bioactive terpenes (caryophyllene oxide, citral, limonene and carvone) was evaluated, following the death mechanism induced on T. cruzi cells. The results were expressed as Inhibitory Concetration 50 (IC50) or Cytotoxic Concentration (CC50) by sigmoidal regression using the statistical software MsxlfitTM. Optical and fluorescent microscopy, flow cytometry, and DNA electrophoresis were used for characterization of the cell death phenotype. The statistical analysis was made by Welch’s ANOVA (software SPSS 15.0). In the conditions tested, the L. alba EOs and their terpenes exhibed a different tripanocidal performance on the diverse parasites stages (p=0.000). Citral chemotype oils evidenced lower IC50 (Epimastigotes: IC50 13,6±1,5μg/mL; Trypomastigotes: IC50 21,9±1,4 μg/mL, and Amastigotes: IC50 74,1±4,4μg/mL) than Carvone chemotype EOs (Epimastigotes: IC50 88,2±3,7μg/mL; Trypomastigotes: IC50 44,9±2,5μg/mL, and Amastigotes: IC50 >150μg/mL). The better selectivity index (SI) was achieved by the monoterpene limonene (Epimastigotes: SI=7,1; Trypomastigotes: SI=32,8, and Amastigotes: SI=10,3), with a CC50 value on Vero cells of 297,1±2,4μg/mL. Limonene presented in vitro synergistic interaction with Benznidazole (with a Fractional Inhibitory Concentration (FIC) of 0,44±0,13, 0,42±0,10 and 0,58±0,13μg/mL for epimastigotes, trypomastigotes, and intracellular amastigotes, respectively), and with caryophyllene oxide (Epimastigotes: FIC=0,49; Tripomastigotes: FIC=0,45; Amastigotes: FIC=0,71). The cellular analysis suggested that these oils or their bioactive terpenes (caryophyllen oxide/limonene) could be inducing cell death by an apoptotic like mechanisms on T. cruzi.
dc.languagespa
dc.publisherBucaramanga : Universidad de Santander, 2017
dc.publisherFacultad Ciencias de la Salud
dc.publisherMaestría en Investigación en enfermedades Infecciosas
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dc.rightsinfo:eu-repo/semantics/openAccess
dc.rightsAtribución-NoComercial 4.0 Internacional (CC BY-NC 4.0)
dc.rightshttps://creativecommons.org/licenses/by-nc/4.0/
dc.rightsDerechos Reservados - Universidad de Santander, 2017
dc.titleCaracterización de la actividad tripanocida y citotóxica de aceites esenciales de lippia alba y sus terpenos bioactivos sobre trypanosoma cruzi
dc.typeTrabajo de grado - Maestría


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