Holothuria leucospilota Extract Induces Apoptosis in Leishmania major Promastigotes
Background: The present study aimed to survey antileishmanial activity of methanolic Holothuria leucospilota extract against Leishmania major promastigotes in vitro. Methods: Promastigotes were cultured in RPMI 1640 and after reaching the stationary phase, the study was conducted with different co...
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Tehran University of Medical Sciences
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doaj-ded832f9cd0141a09ae53d952738a7642021-04-02T15:27:48ZengTehran University of Medical SciencesIranian Journal of Parasitology1735-70202008-238X2016-12-01113592Holothuria leucospilota Extract Induces Apoptosis in Leishmania major PromastigotesMasoud FOROUTAN-RAD0Shahram KHADEMVATAN1Jasem SAKI2Mahmoud HASHEMITABAR3Dept. of Medical Parasitology, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, IranCellular and Molecular Research Center & Department of Medical Parasitology and Mycology, Urmia University of Medical Sciences, Urmia, IranDept. of Medical Parasitology, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran AND Cellular and Molecular Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, IranCellular and Molecular Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran Background: The present study aimed to survey antileishmanial activity of methanolic Holothuria leucospilota extract against Leishmania major promastigotes in vitro. Methods: Promastigotes were cultured in RPMI 1640 and after reaching the stationary phase, the study was conducted with different concentrations of the extract. Afterwards, MTT colorimetric assay for the obtaining of 50% inhibitory concentration (IC50) was utilized. Furthermore, in order to determine the possible induction of apoptosis in L. major promastigotes, flow cytometry and DNA fragmentation methods were employed using annexin-V FLUOS staining kit and DNA ladder kit, respectively. Results: The IC50 value of H. leucospilota extract at three time points of 24, 48, and 72 h was estimated 2000, 300 and 85 µg/ml, respectively. In addition, the extract revealed a dose and time-dependent antileishmanial activity. Furthermore, various characteristics of apoptosis appeared after L. major promastigotes treatment, which included cell shrinkage, formation of apoptotic bodies, blebbing of the cell membrane, and externalization of phosphatidylserine, although no laddering pattern was observed. Conclusion: The methanolic extract of H. leucospilota possesses lethal effect on L. major promastigotes and induces the apoptosis in parasites. Further studies are required to address the apoptosis mechanism in vivo. https://ijpa.tums.ac.ir/index.php/ijpa/article/view/1204Leishmania majorSea cucumberHolothuria leucospilotaApoptosisMTTFlow cytomtry |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Masoud FOROUTAN-RAD Shahram KHADEMVATAN Jasem SAKI Mahmoud HASHEMITABAR |
spellingShingle |
Masoud FOROUTAN-RAD Shahram KHADEMVATAN Jasem SAKI Mahmoud HASHEMITABAR Holothuria leucospilota Extract Induces Apoptosis in Leishmania major Promastigotes Iranian Journal of Parasitology Leishmania major Sea cucumber Holothuria leucospilota Apoptosis MTT Flow cytomtry |
author_facet |
Masoud FOROUTAN-RAD Shahram KHADEMVATAN Jasem SAKI Mahmoud HASHEMITABAR |
author_sort |
Masoud FOROUTAN-RAD |
title |
Holothuria leucospilota Extract Induces Apoptosis in Leishmania major Promastigotes |
title_short |
Holothuria leucospilota Extract Induces Apoptosis in Leishmania major Promastigotes |
title_full |
Holothuria leucospilota Extract Induces Apoptosis in Leishmania major Promastigotes |
title_fullStr |
Holothuria leucospilota Extract Induces Apoptosis in Leishmania major Promastigotes |
title_full_unstemmed |
Holothuria leucospilota Extract Induces Apoptosis in Leishmania major Promastigotes |
title_sort |
holothuria leucospilota extract induces apoptosis in leishmania major promastigotes |
publisher |
Tehran University of Medical Sciences |
series |
Iranian Journal of Parasitology |
issn |
1735-7020 2008-238X |
publishDate |
2016-12-01 |
description |
Background: The present study aimed to survey antileishmanial activity of methanolic Holothuria leucospilota extract against Leishmania major promastigotes in vitro.
Methods: Promastigotes were cultured in RPMI 1640 and after reaching the stationary phase, the study was conducted with different concentrations of the extract. Afterwards, MTT colorimetric assay for the obtaining of 50% inhibitory concentration (IC50) was utilized. Furthermore, in order to determine the possible induction of apoptosis in L. major promastigotes, flow cytometry and DNA fragmentation methods were employed using annexin-V FLUOS staining kit and DNA ladder kit, respectively.
Results: The IC50 value of H. leucospilota extract at three time points of 24, 48, and 72 h was estimated 2000, 300 and 85 µg/ml, respectively. In addition, the extract revealed a dose and time-dependent antileishmanial activity. Furthermore, various characteristics of apoptosis appeared after L. major promastigotes treatment, which included cell shrinkage, formation of apoptotic bodies, blebbing of the cell membrane, and externalization of phosphatidylserine, although no laddering pattern was observed.
Conclusion: The methanolic extract of H. leucospilota possesses lethal effect on L. major promastigotes and induces the apoptosis in parasites. Further studies are required to address the apoptosis mechanism in vivo.
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topic |
Leishmania major Sea cucumber Holothuria leucospilota Apoptosis MTT Flow cytomtry |
url |
https://ijpa.tums.ac.ir/index.php/ijpa/article/view/1204 |
work_keys_str_mv |
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