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Miltefosine-Induced Apoptotic Cell Death on Leishmania major and L. tropica Strains

Korean Journal of Parasitology 2011년 49권 1호 p.17 ~ 23
 ( Khademvatan Shahram ) - Iran Ahvaz Jundishapur University of Medical Sciences Department of Medical Parasitology

 ( Gharavi Mohammad Javad ) - Tehran University of Medical Sciences Faculty of Allied Medicine Department of Medical Parasitology and Mycology
 ( Rahim Fakher ) - Iran Ahvaz Jundishapur University of Medical Sciences Physiology Research Center
 ( Saki Jasem ) - Iran Ahvaz Jundishapur University of Medical Sciences Department of Medical Parasitology

Abstract


The aim of this study was to assess the cytotoxic effects of various concentrations of miltefosine on Leishmania major (MRHO/IR/75/ER) and L. tropica (MHOM/IR/02/Mash10) promastigotes and to observe the programmed cell death features. The colorimetric MTT assay was used to find L. major and L. tropica viability and the obtained results were expressed as 50% inhibitory concentration (IC50). Also, 50% effective doses (ED50) for L. major and L. tropica amastigotes were also determined. Annexin-V FLUOS staining was performed to study the cell death properties of miltefosine using FACS analysis. Qualitative analysis of the total genomic DNA fragmentation was performed by agarose gel electrophoresis. Furthermore, to observe changes in cell morphology, promastigotes were examined using light microscopy. In both strains of L. major and L. tropica, miltefosine induced dose-dependent death with features of apoptosis, including cell shrinkage, DNA laddering, and externalization of phosphatidylserine. The IC50 was achieved at 22 μM and 11 μM for L. major and L. tropica after 48 hr of incubation, respectively. ED50 of L. major and L. tropica amastigotes were 5.7 μM and 4.2 μM, respectively. Our results indicate that miltefosine induces apoptosis of the causative agent of cutaneous leishmaniasis in a dose-dependent manner. Interestingly, L. major did not display any apoptotic changes when it was exposed to miltefosine in concentrations sufficient to kill L. tropica.

키워드

Leishmania major; Leishmania tropica; miltefosine; apoptosis; IC50; ED50
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