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http://dx.doi.org/10.3347/kjp.2011.49.1.l7

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

Khademvatan, Shahram (Department of Medical Parasitology, Ahvaz Jundishapur University of Medical Sciences)
Gharavi, Mohammad Javad (Department of Medical Parasitology and Mycology, Faculty of Allied Medicine, Tehran University of Medical Sciences)
Rahim, Fakher (Cellular and Molecular Research Center, Ahvaz Jundishapur University of Medical Sciences)
Saki, Jasem (Department of Medical Parasitology, Ahvaz Jundishapur University of Medical Sciences)
Publication Information
Parasites, Hosts and Diseases / v.49, no.1, 2011 , pp. 17-23 More about this Journal
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 FAGS 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 ${\mu}M$ and 11 ${\mu}M$ for L. major and L. tropica after 48 hr of incubation, respectively. ED50 of L. major and L. tropica amastigotes were 5.7 ${\mu}M$ and 4.2 ${\mu}M$, 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.
Keywords
Leishmania major; Leishmania tropica; miltefosine; apoptosis; IC50; ED50;
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1 Hadighi R, Boucher P, Khamesipour A, Meamar AR, Roy G, Ouellette M, Mohebali M. Glucantime-resistant Leishmania tropica isolated from Iranian patients with cutaneous leishmaniasis are sensitive to alternative antileishmania drugs. Parasitol Res 2007; 101: 1319-1322.   DOI   ScienceOn
2 Esmaeili J, Mohebali M, Edrissian GH, Rezayat SM, Ghazi-Khansari M, Charehdar S. Evaluation of miltefosine against L. major (mrho/ir/75/er): in vitro and in vivo studies. Acta Med Iran 2008; 4: 191-196.
3 Arnoult D, Akarid K, Grodet A, Petit PX, Estaquier J, Ameisen JC. On the evolution of programmed cell death: apoptosis of the unicellular eukaryote Leishmania major involves cysteine proteinase activation and mitochondrion permeabilization. Cell Death Differ 2002; 9: 65-81.   DOI   ScienceOn
4 Paris C, Loiseau PM, Bories C, Breard J. Miltefosine induces apoptosis-like death in Leishmania donovani promastigotes. Antimicrob Agents Chemother 2004; 48: 852-859.   DOI   ScienceOn
5 Wanderley JL, Barcinski MA. Apoptosis and apoptotic mimicry: the Leishmania connection. Cell Mol Life Sci 2010; 67: 1653-1659.   DOI   ScienceOn
6 Khademvatan S, Gharavi MJ, Akhlaghi L, Samadikuchaksaraei A, Oormazdi H, Mousavizadeh K, Hadighi R, Saki J. Induction of apoptosis by miltefosine in Iranian strain of Leishmania infantum promastigotes. Iranian J Parasitol 2009; 4: 23-31.
7 Lee N, Bertholet S, Debrabant A, Muller J, Duncan R, Nakhasi HL. Programmed cell death in the unicellular protozoan parasite Leishmania. Cell Death Differ 2002; 9: 53-64.   DOI   ScienceOn
8 Mukherjee SB, Das M, Sudhandiran G, Shaha C. Increase in cytosolic $Ca^{2+}$ levels through the activation of non-selective cation channels induced by oxidative stress causes mitochondrial depolarization leading to apoptosis-like death in Leishmania donovani promastigotes. J Biol Chem 2002; 277: 24717-24727.   DOI
9 Mohebali M, Edrissian GH, Nadim A, Hajjaran H, Akhoundi B, Hooshmand B, Zarei Z, Arshi A, Mirsamadi N, Naeini KM, Mamishi S, Sanati AA, Moshfe AA, Charehdar S, Fakhar M. Application of direct agglutination test (DAT) for the diagnosis and seroepidemiological studies of visceral leishmaniasis in the Islamic Republic of Iran. Iran J Parasitol 2006; 1: 15-25.
10 Momeni AZ, Aminjavaheri M. Successful treatment of non-healing cases of cutaneous leishmaniasis, using a combination of meglumine antimoniate plus allopurinol. Eur J Dermatol 2003; 13: 40-43.
11 Hadighi R, Mohebali M, Boucher P, Hajjaran H, Khamesipour A, Ouellette M. Unresponsiveness to Glucantime treatment in Iranian cutaneous leishmaniasis due to drug-resistant Leishmania tropica parasites. PLoS Med 2006; 3: e162.   DOI
12 Mishra BB, Kale RR, Singh RK, Tiwari VK. Alkaloids: future prospective to combat leishmaniasis. Fitoterapia 2009; 80: 81-90.   DOI   ScienceOn
13 Richard JV, Werbovetz KA. New antileishmanial candidates and lead compounds. Curr Opin Chem Biol 2010; 14: 447-455.   DOI   ScienceOn
14 Mohebali M, Fotouhi A, Hooshmand B, Zarei Z, Akhoundi B, Rahnema A, Razaghian AR, Kabir MJ, Nadim A. Comparison of miltefosime and meglumine antimoniate for the treatment of zoonotic cutaneous leishmaniasis (ZCL) by a randomized clinical trial in Iran. Acta Trop 2007; 103: 33-40.   DOI   ScienceOn
15 Verma NK, Dey CS. Possible mechanism of miltefosine-mediated death of Leishmania donovani. Antimicrob Agents Chemother 2004; 48: 3010-3015.   DOI   ScienceOn
16 Wortmann G, Hochberg L, Houng HH, Sweeney C, Zapor M, Aronson N, Weina P, Ockenhouse CF. Rapid identification of Leishmania complexes by a real-time PCR assay. Am J Trop Med Hyg 2005; 73: 999-1004.
17 Vaux DL. Toward an understanding of the molecular mechanisms of physiological cell death. Proc Natl Acad Sci USA 1993; 90: 786-789.   DOI   ScienceOn
18 Ameen M. Cutaneous and mucocutaneous leishmaniasis: emerging therapies and progress in disease management. Expert Opin Pharmacother 2010; 11: 557-569.   DOI   ScienceOn
19 Ellis RE, Yuan JY, Horvitz HR. Mechanisms and functions of cell death. Ann Rev Cell Biol 1991; 7: 663-698.   DOI   ScienceOn
20 Raff MC. Social controls on cell survival and cell death. Nature 1992; 356(6368): 397-400.   DOI   ScienceOn
21 Steller H. Mechanisms and genes of cellular suicide. Science 1995; 267(5203): 1445-1449.   DOI
22 Heussler VT, Küenzi P, Rottenberg S. Inhibition of apoptosis by intracellular protozoan parasites. Int J Parasitol 2001; 31: 1166-1176.   DOI   ScienceOn
23 McConkey DJ, Zhivotovsky B, Orrenius S. Apoptosis--molecular mechanisms and biomedical implications. Mol Aspects Med 1996; 17: 1-110.   DOI   ScienceOn
24 Hamann A, Brust D, Osiewacz HD. Apoptosis pathways in fungal growth, development and ageing. Trends Microbiol 2008; 16: 276-283.   DOI   ScienceOn
25 Wanderley JL, Benjamin A, Real F, Bonomo A, Moreira ME, Barcinski MA. Apoptotic mimicry: an altruistic behavior in host/Leishmania interplay. Braz J Med Biol Res 2005; 38: 807-812.   DOI
26 Guerin PJ, Olliaro P, Sundar S, Boelaert M, Croft SL, Desjeux P, Wasunna MK, Bryceson AD. Visceral leishmaniasis: current status of control, diagnosis, and treatment, and a proposed research and development agenda. Lancet Infect Dis 2002; 2: 494-501.   DOI   ScienceOn
27 Mohebali M, Javadian E, Yaghoobi-Ershadi MR, Akhavan AA, Hajjaran H, Abaei MR. Characterization of Leishmania infection in rodents from endemic areas of the Islamic Republic of Iran. East Mediterr Health J 2004; 10: 591-599.