title: Diospyros perigrena bark extract induced apoptosis in filarial parasite Setaria cervi through generation of reactive oxygen species.
abstract: BACKGROUND
  Lymphatic filariasis is a major neglected tropical disease. Diospyros perigrena Gurke (Ebenaceae) was selected for antifilarial chemotherapy because of unavailability of proper medicine. In India, different parts of this plant were used for the treatment of diabetes, diarrhea, dysentery, cholera, mouth ulcers, and wounds.
  
  
  OBJECTIVE
  The present study was undertaken to access antifilarial potential and mechanism of action of n-butanol extract (NBE) of D. perigrena stem bark on Setaria cervi Rudolphi (Onchocercidae).
  
  
  METHODS
  In vitro efficacy and apoptotic mechanism were evaluated by Hoechst, TUNEL, DNA fragmentation assay, pro- and anti-apoptotic gene expression in NBE (250, 125, 62.5, 31.25, and 15.6 µg/ml)-treated S. cervi after 24 h of incubation. Reactive oxygen species (ROS) up-regulation was also determined by GSH, GST, SOD assays, and super oxide anion level.
  
  
  RESULTS
  Significant in vitro antifilarial activity of NBE was found 50% inhibitory concentration (IC50): adult = 57.6 μg/ml, microfilariae (mf) = 56.1 μg/ml, and lethal dose (LD100) in mf is 187.17 μg/ml) after 24 h of treatment. NBF-induced apoptosis was proved by Hoechst, TUNEL, RT-PCR, and Western blot method. NBF (250 µg/ml) decreased the level of GSH (17.8%) and GST (65.4%), increased SOD activity (1.42-fold) and super oxide anion production (1.32-fold) in the treated parasite which culminated into ROS up-regulation.
  
  
  CONCLUSIONS
  NBE induced apoptosis in different life cycle stages of S. cervi. In future, a detailed study of NBF will give us a novel antifilarial compound which will be used for antifilarial chemotherapy.
authors.affiliation: Parasitology Laboratory, Department of Zoology, Center for Advanced Studies, Visva-Bharati University , Santiniketan, West Bengal , India and.
authors.forename: Prasanta
authors.initials: P
authors.lastname: Saini
authors.forename: Niladri
authors.initials: N
authors.lastname: Mukherjee
authors.forename: Suprabhat
authors.initials: S
authors.lastname: Mukherjee
authors.forename: Priya
authors.initials: P
authors.lastname: Roy
authors.forename: Prajna
authors.initials: P
authors.lastname: Gayen
authors.forename: Deepak
authors.initials: D
authors.lastname: Kumar
authors.forename: Bikas Chandra
authors.initials: BC
authors.lastname: Pal
authors.forename: Santi P Sinha
authors.initials: SP
authors.lastname: Babu
mesh_terms: D020001:1-Butanol; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D006690:Bisbenzimidazole; D004396:Coloring Agents; D004247:DNA / Q000187:drug effects; D029788:Diospyros / Q000737:chemistry*; D005368:Filariasis / Q000188:drug therapy / Q000523:psychology; D005369:Filaricides / Q000494:pharmacology*; D020287:In Situ Nick-End Labeling; D024301:Plant Bark / Q000737:chemistry*; D010936:Plant Extracts / Q000494:pharmacology*; D017382:Reactive Oxygen Species / Q000378:metabolism*; D017182:Setaria Nematode / Q000187:drug effects* / Q000378:metabolism; D012997:Solvents; D013778:Tetrazolium Salts; D013844:Thiazoles
chemical_list: D004396:Coloring Agents; D005369:Filaricides; D010936:Plant Extracts; D017382:Reactive Oxygen Species; D012997:Solvents; D013778:Tetrazolium Salts; D013844:Thiazoles; D020001:1-Butanol; D004247:DNA; C022616:thiazolyl blue; D006690:Bisbenzimidazole
keywords: Antifilarial; DNA fragmentation; cell death abnormal; glutathione; relative movability; superoxide dismutase