title: Antifilarial effect of ursolic acid from Nyctanthes arbortristis: molecular and biochemical evidences.
abstract: A bio-assay guided fractionation and purification approach was used to examine in vitro antifilarial activities of the crude methanolic extract of Nyctanthes arbortristis as well as fractions and isolated compound. From ethyl-acetate fraction we isolated and identified a triterpenoid compound which has been characterized as ursolic acid (UA) by HPLC and NMR data. We are reporting for the first time isolation and identification of UA from the leaves of N. arbortristis. The crude extract and UA showed significant micro- as well as macrofilaricidal activities against the oocyte, microfilaria and adult of Setaria cervi (S. cervi) by dye exclusion test and MTT reduction assay. Significant microfilaricidal activity of UA was further proved against mf of W. bancrofti by viability assay. The findings thus provide a new lead for development of a suitable filaricide from natural products. The molecular mechanism of UA was investigated by performing TUNEL, Hoechst staining, Annexin V-Cy3, flow cytometric analysis and DNA fragmentation assay. Differential expressions of pro- and anti-apoptotic genes were observed at the transcription and translational levels in a dose-dependent manner. Depletion in the worm GSH level and elevation in the parasite GST, SOD and super oxide anion indicated the generation of ROS. In this investigation we are reporting for the first time that UA acts its antifilarial effect through induction of apoptosis and by downregulating and altering the level of some key antioxidants like GSH, GST and SOD of S. cervi.
authors.affiliation: Parasitology Laboratory, Department of Zoology, Centre for Advanced Studies, Visva-Bharati University, Santiniketan 731235, 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 731235, West Bengal, India.
authors.forename: Prajna
authors.initials: P
authors.lastname: Gayen
authors.affiliation: National Institute of Pharmaceutical Education and Research (NIPER), 4, Raja S.C. Mullick Road, Jadavpur, Kolkata 700032, West Bengal, India.
authors.forename: Deepak
authors.initials: D
authors.lastname: Kumar
authors.affiliation: Parasitology Laboratory, Department of Zoology, Centre for Advanced Studies, Visva-Bharati University, Santiniketan 731235, West Bengal, India.
authors.forename: Ananya
authors.initials: A
authors.lastname: Nayak
authors.affiliation: Parasitology Laboratory, Department of Zoology, Centre for Advanced Studies, Visva-Bharati University, Santiniketan 731235, 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 731235, West Bengal, India.
authors.forename: Suprabhat
authors.initials: S
authors.lastname: Mukherjee
authors.affiliation: National Institute of Pharmaceutical Education and Research (NIPER), 4, Raja S.C. Mullick Road, Jadavpur, Kolkata 700032, West Bengal, India.
authors.forename: Bikas C
authors.initials: BC
authors.lastname: Pal
authors.affiliation: Parasitology Laboratory, Department of Zoology, Centre for Advanced Studies, Visva-Bharati University, Santiniketan 731235, West Bengal, India. Electronic address: spsinhababu@gmail.com.
authors.forename: Santi P Sinha
authors.initials: SP
authors.lastname: Babu
mesh_terms: D000328:Adult; D000818:Animals; D015153:Blotting, Western; D004305:Dose-Response Relationship, Drug; D005260:Female; D005369:Filaricides / Q000737:chemistry / Q000494:pharmacology*; D005978:Glutathione; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008297:Male; D029592:Oleaceae / Q000737:chemistry*; D018384:Oxidative Stress / Q000187:drug effects; D018515:Plant Leaves / Q000737:chemistry; D010946:Plants, Medicinal; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017182:Setaria Nematode / Q000187:drug effects*; D013482:Superoxide Dismutase; D013481:Superoxides; D014315:Triterpenes / Q000737:chemistry / Q000494:pharmacology*; D014958:Wuchereria bancrofti / Q000187:drug effects*; D000097245:Ursolic Acid
chemical_list: D005369:Filaricides; D005978:Glutathione; D005982:Glutathione Transferase; D013482:Superoxide Dismutase; D013481:Superoxides; D014315:Triterpenes; D000097245:Ursolic Acid
keywords: Apoptosis; Nyctanthes arbortristis; Reactive oxygen species; Setaria cervi; Ursolic acid; Wuchereria bancrofti