Anti-proliferative synergy of lysophospholipid analogues and ketoconazole against Trypanosoma cruzi (Kinetoplastida: Trypanosomatidae): cellular and ultrastructural analysis.
OBJECTIVE
Investigation of the antiproliferative synergy of the lysophospholipid analogues (LPAs) edelfosine, ilmofosine and miltefosine with the ergosterol biosynthesis inhibitor ketoconazole against Trypanosoma cruzi.


METHODS
The effect of LPAs, ketoconazole and their combination was evaluated against epimastigotes and intracellular amastigotes by the parameter IC50 leading to construction of isobolograms, for determination of a synergic effect. For epimastigotes, ultrastructural damage induced by these treatments was evaluated by transmission and scanning electron microscopy.


RESULTS
Synergy was confirmed against both epimastigotes and amastigotes of the parasite. Edelfosine or ketoconazole alone induced morphological alterations in the plasma membrane and reservosomes of the parasites, while in combination, they also led to severe mitochondrial damage, formation of autophagic structures and multinucleation. Scanning electron microscopy confirmed the effect at the plasma membrane and also revealed alterations in the shape of the parasites.


CONCLUSIONS
Our results describe the synergic anti-proliferative effect of LPAs and ketoconazole against epimastigotes and intracellular amastigotes and suggest that in epimastigotes, plasma membrane, reservosomes and mitochondria are targets of these drugs, possibly by interference with lipid metabolism.
Santa-Rita
Ricardo M
RM
Departmento de Ultra-estrutura e Biologia Celular, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, CP 926, 21045-900, Rio de Janeiro, Brazil.
Lira
Renee
R
Barbosa
Helene Santos
HS
Urbina
Julio A
JA
de Castro
Solange L
SL
D000818:Animals; D002522:Chlorocebus aethiops; D004357:Drug Synergism; D004359:Drug Therapy, Combination; D004875:Ergosterol / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D007654:Ketoconazole / Q000494:pharmacology*; D008246:Lysophospholipids / Q000737:chemistry / Q000494:pharmacology*; D008854:Microscopy, Electron; D008855:Microscopy, Electron, Scanning; D021261:Parasitic Sensitivity Tests / Q000379:methods; D010742:Phospholipid Ethers / Q000494:pharmacology*; D010767:Phosphorylcholine / Q000031:analogs & derivatives* / Q000494:pharmacology*; D014344:Trypanocidal Agents / Q000494:pharmacology*; D014349:Trypanosoma cruzi / Q000187:drug effects / Q000254:growth & development* / Q000648:ultrastructure; D014709:Vero Cells
D008246:Lysophospholipids; D010742:Phospholipid Ethers; D014344:Trypanocidal Agents; D010767:Phosphorylcholine; C026659:edelfosine; C039128:miltefosine; D007654:Ketoconazole; D004875:Ergosterol