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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Main Authors: Masoud FOROUTAN-RAD, Shahram KHADEMVATAN, Jasem SAKI, Mahmoud HASHEMITABAR
Format: Article
Language:English
Published: Tehran University of Medical Sciences 2016-12-01
Series:Iranian Journal of Parasitology
Subjects:
MTT
Online Access:https://ijpa.tums.ac.ir/index.php/ijpa/article/view/1204
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spelling 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.
topic Leishmania major
Sea cucumber
Holothuria leucospilota
Apoptosis
MTT
Flow cytomtry
url https://ijpa.tums.ac.ir/index.php/ijpa/article/view/1204
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