title: Antifilarial effects of polyphenol rich ethanolic extract from the leaves of Azadirachta indica through molecular and biochemical approaches describing reactive oxygen species (ROS) mediated apoptosis of Setaria cervi.
abstract: Lymphatic filariasis, a global cause of morbidity needs much more attention in developing potent therapeutics that can be effective against both microfilariae (mf) and adults. Efficient botanicals that can induce apoptosis of filarial parasites possibly can provide a direction towards developing new class of antifilarials. In this work we have evaluated the antifilarial efficacy of an optimized polyphenol rich ethanolic extract of Azadirachta indica leaves (EEA). A. indica A. Juss has been widely used in the traditional Indian medicinal system 'Ayurveda' for the treatment of a variety of ailments. A thorough investigation towards biochemical and molecular mechanisms describing ROS mediated apoptosis in Setaria cervi was performed. Motility reduction, MTT reduction assay and dye exclusion test have confirmed the micro- and macrofilaricidal potential of EEA. Alterations were visible in mf and trichrome stained section of EEA-treated adult worms. We have found cellular disturbances in EEA-treated parasites characterized by chromatin condensation, in situ DNA fragmentation and nucleosomal DNA laddering. Depletion in worm GSH level and elevation in parasite GST, SOD, catalase, GPx and superoxide anion indicated the generation of ROS. Our results provided experimental evidence supporting that EEA causes a decreased expression of anti-apoptotic genes and increased pro-apoptotic gene expression at the level of both transcription and translation. Here we are reporting for the first time that antifilarial activity of EEA is mediated by ROS up regulation and apoptosis.
authors.affiliation: Parasitology Laboratory, Department of Zoology, Centre for Advanced Studies, Visva-Bharati University, Santiniketan 731 235, West Bengal, India.
authors.forename: Niladri
authors.initials: N
authors.lastname: Mukherjee
authors.affiliation: Parasitology Laboratory, Department of Zoology, Centre for Advanced Studies, Visva-Bharati University, Santiniketan 731 235, West Bengal, India.
authors.forename: Suprabhat
authors.initials: S
authors.lastname: Mukherjee
authors.affiliation: Parasitology Laboratory, Department of Zoology, Centre for Advanced Studies, Visva-Bharati University, Santiniketan 731 235, West Bengal, India.
authors.forename: Prasanta
authors.initials: P
authors.lastname: Saini
authors.affiliation: Parasitology Laboratory, Department of Zoology, Centre for Advanced Studies, Visva-Bharati University, Santiniketan 731 235, West Bengal, India.
authors.forename: Priya
authors.initials: P
authors.lastname: Roy
authors.affiliation: Parasitology Laboratory, Department of Zoology, Centre for Advanced Studies, Visva-Bharati University, Santiniketan 731 235, West Bengal, India. Electronic address: spsinhababu@gmail.com.
authors.forename: Santi P
authors.initials: SP
authors.lastname: Sinha Babu
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D017209:Apoptosis / Q000235:genetics; D031601:Azadirachta / Q000737:chemistry*; D002417:Cattle; D053938:DNA Fragmentation / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D005260:Female; D005369:Filaricides / Q000302:isolation & purification / Q000494:pharmacology*; D005786:Gene Expression Regulation / Q000187:drug effects; D007559:Ivermectin / Q000494:pharmacology; D008297:Male; D010084:Oxidation-Reduction / Q000187:drug effects; D010936:Plant Extracts / Q000302:isolation & purification / Q000494:pharmacology*; D018515:Plant Leaves / Q000737:chemistry; D059808:Polyphenols / Q000302:isolation & purification / Q000494:pharmacology*; D017382:Reactive Oxygen Species / Q000378:metabolism*; D017182:Setaria Nematode / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism
chemical_list: D005369:Filaricides; D010936:Plant Extracts; D059808:Polyphenols; D017382:Reactive Oxygen Species; D007559:Ivermectin
keywords: 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl-tetrazolium bromide; ANOVA; Apaf; Apoptosis; Azadirachta indica; B-cell lymphoma 2; BCl-2; CED; CM; CP; DMSO; EEA; EGL; EtBr; EtOH; FBS; GPx; GSH; GST; MTT; NBT; OPD; Optimized extraction; PBS; PVDF; RM; ROS; RSM; RT-PCR; Reactive oxygen species (ROS); SOD; Setaria cervi; TPC; TUNEL; Terminal deoxynucleotidyl transferase dUTP nick end labeling; analysis of variance; apoptotic protease activating factor; cell-death abnormal; crude powder; culture media; dimethyl sulfoxide; egg laying defective; ethanol; ethanolic extract of Azadirachta indica leaves; ethidium bromide; foetal bovine serum; glutathione; glutathione peroxidase; glutathione-S-transferase; mf; microfilariae; nitroblue tetrazolium; ortho-phenylenediamine; p53; phosphate buffered saline; poly vinylidene difluoride; protein 53; reactive oxygen species; relative motility; response surface methodology; reverse transcriptase polymerase chain reaction; superoxide dismutase; total polyphenolic contents