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Biological approaches to characterize the mode of action of two 5-nitroindazolinone prototypes on Trypanosoma cruzi bloodstream trypomastigotes

Published online by Cambridge University Press:  17 June 2016

CRISTINA FONSECA-BERZAL ,
CRISTIANE FRANÇA DA SILVA ,
RUBEM F. S. MENNA-BARRETO ,
MARCOS MEUSER BATISTA ,
JOSÉ A. ESCARIO ,
VICENTE J. ARÁN ,
ALICIA GÓMEZ-BARRIO  and
MARIA DE NAZARÉ C. SOEIRO
CRISTINA FONSECA-BERZAL*
Affiliation:
CEI Campus Moncloa, UCM-UPM & CSIC, Madrid, Spain Departamento de Parasitología, Facultad de Farmacia, Universidad Complutense de Madrid, Pza. Ramón y Cajal s/n, 28040 Madrid, Spain
CRISTIANE FRANÇA DA SILVA
Affiliation:
Laboratório de Biologia Celular, Instituto Oswaldo Cruz, Fiocruz, Av. Brasil 4365, 21040-900 Rio de Janeiro, Brazil
RUBEM F. S. MENNA-BARRETO
Affiliation:
Laboratório de Biologia Celular, Instituto Oswaldo Cruz, Fiocruz, Av. Brasil 4365, 21040-900 Rio de Janeiro, Brazil
MARCOS MEUSER BATISTA
Affiliation:
Laboratório de Biologia Celular, Instituto Oswaldo Cruz, Fiocruz, Av. Brasil 4365, 21040-900 Rio de Janeiro, Brazil
JOSÉ A. ESCARIO
Affiliation:
CEI Campus Moncloa, UCM-UPM & CSIC, Madrid, Spain Departamento de Parasitología, Facultad de Farmacia, Universidad Complutense de Madrid, Pza. Ramón y Cajal s/n, 28040 Madrid, Spain
VICENTE J. ARÁN
Affiliation:
CEI Campus Moncloa, UCM-UPM & CSIC, Madrid, Spain Instituto de Química Médica (IQM), Consejo Superior de Investigaciones Científicas (CSIC), c/Juan de la Cierva 3, 28006 Madrid, Spain
ALICIA GÓMEZ-BARRIO
Affiliation:
CEI Campus Moncloa, UCM-UPM & CSIC, Madrid, Spain Departamento de Parasitología, Facultad de Farmacia, Universidad Complutense de Madrid, Pza. Ramón y Cajal s/n, 28040 Madrid, Spain
MARIA DE NAZARÉ C. SOEIRO
Affiliation:
Laboratório de Biologia Celular, Instituto Oswaldo Cruz, Fiocruz, Av. Brasil 4365, 21040-900 Rio de Janeiro, Brazil
*
*Corresponding author: Departamento de Parasitología, Facultad de Farmacia, Universidad Complutense de Madrid, Pza. Ramón y Cajal s/n, 28040 Madrid, Spain. Tel.: +34 913941818. Fax: +34 3941815. E-mail: crfonseca@pdi.ucm.es
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Summary

The phenotypic activity of two 5-nitroindazolinones, i.e. 2-benzyl-1-propyl (22) and 2-benzyl-1-butyl (24) derivatives, previously proposed as anti-Trypanosoma cruzi prototypes, was presently assayed on bloodstream trypomastigotes (BT) of the moderately drug-resistant Y strain. Further exploration of putative targets and cellular mechanisms involved in their activity was also carried out. Therefore, transmission electron microscopy, high-resolution respirometry and flow cytometry procedures were performed on BT treated for up to 24 h with the respective EC50 value of each derivative. Results demonstrated that although 22 and 24 were not as active as benznidazole in this in vitro assay on BT, both compounds triggered important damages in T. cruzi that lead to the parasite death. Ultrastructural alterations included shedding events, detachment of plasma membrane and nuclear envelope, loss of mitochondrial integrity, besides the occurrence of a large number of intracellular vesicles and profiles of endoplasmic reticulum surrounding cytoplasmic organelles such as mitochondrion. Moreover, both derivatives affected mitochondrion leading to this organelle dysfunction, as reflected by the inhibition in oxygen consumption and the loss of mitochondrial membrane potential. Altogether, the findings exposed in the present study propose autophagic processes and mitochondrial machinery as part of the mode of action of both 5-nitroindazolinones 22 and 24 on T. cruzi trypomastigotes.

Keywords

Chagas diseasetransmission electron microscopyhigh-resolution respirometryflow cytometrymitochondrionautophagy
Type
Research Article
Information
Parasitology , Volume 143 , Issue 11 , September 2016 , pp. 1469 - 1478
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Copyright
Copyright © Cambridge University Press 2016 

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