title: The sesquiterpene (-)-α-bisabolol is active against the causative agents of Old World cutaneous leishmaniasis through the induction of mitochondrial-dependent apoptosis.
abstract: Cutaneous leishmaniasis treatment remains challenging due to the absence of a satisfactory treatment. The screening of natural compounds is a valuable strategy in the search of new drugs against leishmaniasis. The sesquiterpene (-)-α-bisabolol is effective in vivo against visceral leishmaniasis due to Leishmania infantum, but its mechanism of action remains elusive. The aim of this study is to validate this promising compound against the causative species of Old World cutaneous leishmaniasis and to get an insight into its antileishmanial mode of action. The compound was evaluated on L. tropica promastigotes and intracellular amastigotes using bone marrow-derived macrophages and its cytotoxicity was evaluated on L929 fibroblasts. The reactive oxygen species generation was evaluated using a sensitive probe. Mitochondrial depolarization was assessed evaluating the fluorescence due to rhodamine 123 in a flow cytometer. Apoptosis was investigated by measuring the fluorescence due to annexin V and propidium iodide in a flow cytometer. The ultrastructure of treated promastigotes and intracellular amastigotes was analysed through transmission electron microscopy. (-)-α-Bisabolol was active against L. tropica intracellular amastigotes displaying an inhibitory concentration 50 % of 25.2 µM and showing low cytotoxicity. This compound induced time and dose-dependent oxidative stress, mitochondrial depolarization and phosphatidilserine externalization (a marker of apoptosis). These effects were noticed at a low concentration and short exposure time. In the ultrastructural analyses, the treated parasites showed mitochondrial disruption, presence of electron-dense structures and chromatin condensation. These results suggest that this natural compound induces oxidative stress and mitochondrial-dependent apoptosis on Leishmania without disturbing the plasma membrane.
authors.affiliation: Departamento de Parasitología, Facultad de Farmacia, Universidad de Granada, Campus de Cartuja s/n, 18071, Granada, Spain. victorianocl@gmail.com.
authors.forename: V
authors.identifier: 0000-0001-9152-6486
authors.initials: V
authors.lastname: Corpas-López
authors.affiliation: Departamento de Parasitología, Facultad de Farmacia, Universidad de Granada, Campus de Cartuja s/n, 18071, Granada, Spain.
authors.forename: G
authors.initials: G
authors.lastname: Merino-Espinosa
authors.affiliation: Departamento de Parasitología, Facultad de Farmacia, Universidad de Granada, Campus de Cartuja s/n, 18071, Granada, Spain.
authors.forename: V
authors.initials: V
authors.lastname: Díaz-Sáez
authors.affiliation: Departamento de Parasitología, Facultad de Farmacia, Universidad de Granada, Campus de Cartuja s/n, 18071, Granada, Spain.
authors.forename: F
authors.initials: F
authors.lastname: Morillas-Márquez
authors.affiliation: Departamento de Farmacología, Facultad de Farmacia, Universidad de Granada, Campus de Cartuja s/n, 18071, Granada, Spain.
authors.forename: M C
authors.initials: MC
authors.lastname: Navarro-Moll
authors.affiliation: Departamento de Parasitología, Facultad de Farmacia, Universidad de Granada, Campus de Cartuja s/n, 18071, Granada, Spain.
authors.forename: J
authors.initials: J
authors.lastname: Martín-Sánchez
mesh_terms: D000818:Animals; D000981:Antiprotozoal Agents / Q000737:chemistry / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D002460:Cell Line; D006801:Humans; D018314:Leishmania infantum / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D016773:Leishmaniasis, Cutaneous / Q000469:parasitology*; D008264:Macrophages / Q000187:drug effects / Q000378:metabolism; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D051379:Mice; D008928:Mitochondria / Q000187:drug effects* / Q000378:metabolism; D000081222:Monocyclic Sesquiterpenes; D018384:Oxidative Stress / Q000187:drug effects; D017382:Reactive Oxygen Species / Q000378:metabolism; D012717:Sesquiterpenes / Q000737:chemistry / Q000494:pharmacology*
chemical_list: D000981:Antiprotozoal Agents; D000081222:Monocyclic Sesquiterpenes; D017382:Reactive Oxygen Species; D012717:Sesquiterpenes; C004497:bisabolol
keywords: Apoptosis; Cutaneous leishmaniasis; Mitochondrial damage; Oxidative stress; Sesquiterpene