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Amiodarone triggers induction of apoptosis in cutaneous leishmaniasis agents
Somayeh Bahrami1, Shahram Khademvatan2, Mohammad Hossein Razi Jalali1
1a Faculty of Veterinary Medicine, Department of Parasitology , Shahid Chamran University of Ahvaz , Ahvaz , Iran.
Abstract:
Leishmaniasis is a parasitic disease that is an important problem of public health worldwide. The aim of this study was to assess the cytotoxic effects of amiodarone (AMD) on Leishmania tropica (MHOM/AF/88/KK27) and L. major (MRHO/IR/75/ER) promastigotes and to observe the programmed cell death features. The colorimetric MTT assay was used to find L. tropica and L. major viability and the obtained results were expressed as 50% inhibitory concentration (IC50). Annexin-V FLUOS staining was performed to study the cell death properties of AMD using fluorescence-activated cell-sorting 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. The IC50 was achieved at 55 and 81 μM for L. tropica and L. major after 48 h of incubation, respectively. In both strains, AMD induced death with features of apoptosis, including externalization of phosphatidylserine, DNA laddering, and cell shrinkage. Our findings indicate that AMD may induce apoptosis on the causative agents of cutaneous leishmaniasis.
Insights
Amiodarone (AMD) shows cytotoxic effects against Leishmania parasites causing leishmaniasis. This drug induces programmed cell death, offering potential for new cutaneous leishmaniasis treatments.
Area of Science:
- Parasitology
- Pharmacology
- Cell Biology
Background:
- Leishmaniasis is a significant global public health concern caused by Leishmania parasites.
- Current treatments for leishmaniasis have limitations, necessitating the search for novel therapeutic agents.
Purpose of the Study:
- To evaluate the cytotoxic impact of amiodarone (AMD) on Leishmania tropica and Leishmania major promastigotes.
- To investigate the mechanisms of programmed cell death induced by AMD in these parasites.
Main Methods:
- Colorimetric MTT assay to determine parasite viability and calculate 50% inhibitory concentration (IC50).
- Annexin-V FLUOS staining and fluorescence-activated cell sorting (FACS) analysis to assess apoptosis.
- Agarose gel electrophoresis for qualitative analysis of genomic DNA fragmentation.
- Light microscopy for observing morphological changes in promastigotes.
Main Results:
- Amiodarone exhibited inhibitory effects with IC50 values of 55 μM for L. tropica and 81 μM for L. major after 48 hours.
- AMD treatment induced features characteristic of apoptosis in both Leishmania strains.
- Observed apoptotic features included phosphatidylserine externalization, DNA laddering, and cell shrinkage.
Conclusions:
- Amiodarone demonstrates significant cytotoxic activity against Leishmania tropica and Leishmania major.
- The mechanism of AMD-induced cell death involves apoptosis in these parasitic agents.
- These findings suggest amiodarone as a potential candidate for developing new therapies against cutaneous leishmaniasis.
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