Programmed cell death in Acanthamoeba castellanii Neff induced by several molecules present in olive leaf extracts

• DOI: 10.1371/journal.pone.0183795
• Publication Date: 2017-08-31T17:32:11Z
• Authors: Ines Sifaoui; Atteneri López-Ar...; Carmen Mª. Martín...; María Reyes-Batlle; Carolina Wagner; Olfa Chiboub; Mondher Mejri; Basilio Valladares; Manef Abderrabba; José E. Piñero; Jacob Lorenzo-Mor...
• Keywords: Membrane Potential, Mitochondrial; 0301 basic medicine; Acanthamoeba castellanii; Cell Membrane Permeability; Cell Death; Plant Extracts; Science; Q; R; Apoptosis; Triterpenes; 3. Good health; Plant Leaves; 03 medical and health sciences; Adenosine Triphosphate; Acanthamoeba Keratitis; Olea; Ursolic Acid; Medicine; Research Article

Therapy against Acanthamoeba infections such as Granulomatous Amoebic Encephalitis (GAE) and Acanthamoeba Keratitis (AK), remains as an issue to be solved due to the existence of a cyst stage which is highly resistant to most chemical and physical agents. Recently, the activity of Olive Leaf Extracts (OLE) was demonstrated against Acanthamoeba species. However, the molecules involved in this activity were not identified and/or evaluated. Therefore, the aim of this study was to evaluate the activity of the main molecules which are present in OLE and secondly to study their mechanism of action in Acanthamoeba. Among the tested molecules, the observed activities ranged from an IC50 of 6.59 in the case of apigenine to an IC50 > 100 μg/ml for other molecules. After that, elucidation of the mechanism of action of these molecules was evaluated by the detection of changes in the phosphatidylserine (PS) exposure, the permeability of the plasma membrane, the mitochondrial membrane potential and the ATP levels in the treated cells. Vanillic, syringic and ursolic acids induced the higher permeabilization of the plasma membrane. Nevertheless, the mitochondrial membrane was altered by all tested molecules which were also able to decrease the ATP levels to less than 50% in IC90 treated cells after 24 h. Therefore, all the molecules tested in this study could be considered as a future therapeutic alternative against Acanthamoeba spp. Further studies are needed in order to establish the true potential of these molecules against these emerging opportunistic pathogenic protozoa.