Trypanosoma cruzi cell death induced by the Morita-Baylis-Hillman adduct 3-Hydroxy-2-methylene-3-(4-nitrophenylpropanenitrile).
Chagas disease, caused by the protozoan Trypanosoma cruzi, remains a serious health concern due to the lack of effective vaccines or satisfactory treatment. In the search for new compounds against this neglected disease, we have previously demonstrated that the compound 3-Hydroxy-2-methylene-3-(4-nitrophenylpropanenitrile) (MBHA3), derived from the Morita-Baylis-Hillman reaction, effectively caused a loss of viability in both the epimastigote and trypomastigote forms. However, the mechanisms of parasite death elicited by MBHA3 remain unknown. The aim of this study was to better understand the morphophysiological changes and the mechanism of cell death induced by MBHA3 treatment on T. cruzi. To perform this analysis, we used confocal microscopy and flow cytometry to monitor the fluorescent probes such as annexin-V/propidium iodide (AV/PI), calcein-AM/ethidium homodimer (CA/EH), acridine orange (AO) and rhodamine 123 (Rho 123). Lower concentrations of MBHA3 led to alterations in the mitochondrial membrane potential and AO labeling, but did not decrease the viability of the epimastiogote forms, as determined by the CA/EH and AV/PI assays. Conversely, treatment with higher concentrations of MBHA3 led to extensive plasma membrane damage, loss of mitochondrion membrane potential, DNA fragmentation and acidification of the cytoplasm. Our findings suggest that at higher concentrations, MBHA3 induces T. cruzi epimastigote death by necrosis in a mitochondrion-dependent manner.
Departamento de Microbiologia, Centro de Pesquisas Aggeu Magalhães, Recife, PE, Brazil.
Jana M
JM
Sandes
Departamento de Biofísica e Radiobiologia, Universidade Federal de Pernambuco, Recife, PE, Brazil.
Adriana
A
Fontes
Laboratorio de Biologia Parasitaria, Centro Pesquisas Gonçalo Moniz, Salvador, BA, Brazil.
Carlos G
CG
Regis-da-Silva
Departamento de Imunologia, Centro de Pesquisas Aggeu Magalhães, Recife, PE, Brazil.
Maria C A Brelaz
MC
de Castro
LASOM-PB, Departamento de Química, Universidade Federal da Paraíba, João Pessoa, PB, Brazil.
Claudio G
CG
Lima-Junior
LASOM-PB, Departamento de Química, Universidade Federal da Paraíba, João Pessoa, PB, Brazil.
Fábio P L
FP
Silva
LASOM-PB, Departamento de Química, Universidade Federal da Paraíba, João Pessoa, PB, Brazil.
Mário L A A
ML
Vasconcellos
Departamento de Microbiologia, Centro de Pesquisas Aggeu Magalhães, Recife, PE, Brazil.
Regina C B Q
RC
Figueiredo
D000181:Acrylonitrile / Q000031:analogs & derivatives* / Q000494:pharmacology / Q000627:therapeutic use; D001592:Benzyl Alcohols / Q000494:pharmacology* / Q000627:therapeutic use; D016923:Cell Death / Q000187:drug effects*; D002462:Cell Membrane / Q000187:drug effects; D014355:Chagas Disease / Q000188:drug therapy* / Q000469:parasitology; D003593:Cytoplasm / Q000187:drug effects; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D018613:Microscopy, Confocal; D009570:Nitriles; D014349:Trypanosoma cruzi / Q000187:drug effects*
C510493:3-hydroxy-2-methylene-3-(4-nitrophenyl)propanenitrile; D001592:Benzyl Alcohols; D009570:Nitriles; D000181:Acrylonitrile