title: Holothuria leucospilotaExtract Induces Apoptosis inLeishmania majorPromastigotes.
abstract: BACKGROUND
  The present study aimed to survey antileishmanial activity of methanolicHolothuria leucospilotaextract againstLeishmania majorpromastigotes 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 inL. majorpromastigotes, flow cytometry and DNA fragmentation methods were employed using annexin-V FLUOS staining kit and DNA ladder kit, respectively.
  
  
  RESULTS
  The IC50value ofH. leucospilotaextract 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 afterL. majorpromastigotes treatment, which included cell shrinkage, formation of apoptotic bodies, blebbing of the cell membrane, and externalization of phosphatidylserine, although no laddering pattern was observed.
  
  
  CONCLUSIONS
  The methanolic extract ofH. leucospilotapossesses lethal effect onL. majorpromastigotes and induces the apoptosis in parasites. Further studies are required to address the apoptosis mechanism in vivo.
authors.affiliation: Dept. of Medical Parasitology, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.
authors.forename: Masoud
authors.initials: M
authors.lastname: Foroutan-Rad
authors.affiliation: Cellular and Molecular Research Center & Department of Medical Parasitology and Mycology, Urmia University of Medical Sciences, Urmia, Iran.
authors.forename: Shahram
authors.initials: S
authors.lastname: Khademvatan
authors.affiliation: Dept. of Medical Parasitology, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran; Cellular and Molecular Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.
authors.forename: Jasem
authors.initials: J
authors.lastname: Saki
authors.affiliation: Cellular and Molecular Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.
authors.forename: Mahmoud
authors.initials: M
authors.lastname: Hashemitabar
keywords: Apoptosis; DNA Fragmentation; Flow cytomtry; Holothuria leucospilota; Leishmania major; MTT; Sea cucumber