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Antifilarial activity of azadirachtin fuelled through reactive oxygen species induced apoptosis: a thorough molecular study on Setaria cervi

Published online by Cambridge University Press:  23 July 2018

N. Mukherjee ,
N. Joardar  and
S.P. Sinha Babu Open the ORCID record for S.P. Sinha Babu [Opens in a new window]
N. Mukherjee
Affiliation:
Parasitology Laboratory, Department of Zoology (Centre for Advanced Studies), Visva-Bharati University, Santiniketan 731 235, West Bengal, India Cancer Biology & Inflammatory Disorder Division, CSIR - Indian Institute of Chemical Biology, 4- Raja S.C. Mullick Road, Kolkata 700 032, West Bengal, India
N. Joardar
Affiliation:
Parasitology Laboratory, Department of Zoology (Centre for Advanced Studies), Visva-Bharati University, Santiniketan 731 235, West Bengal, India
S.P. Sinha Babu*
Affiliation:
Parasitology Laboratory, Department of Zoology (Centre for Advanced Studies), Visva-Bharati University, Santiniketan 731 235, West Bengal, India
*
Author for correspondence: S.P Sinha Babu E-mail: [email protected]
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Abstract

Efficacious therapeutic strategies against lymphatic filariasis are always sought after. However, natural products are a promising resource for developing effective antifilarial agents. Azadirachtin, a significant tetranortriterpenoid phytocompound found in Azadirachta indica, was evaluated in vitro for antifilarial potential against the filarial parasite Setaria cervi. Dye exclusion and MTT assay confirmed the antifilarial potential of azadirachtin against S. cervi with a median lethal dose (LC50) of 6.28 μg/ml for microfilariae (mf), and 9.55 μg/ml for adult parasites. Morphological aberrations were prominent in the histological sections of the azadirachtin-exposed parasites. Moreover, alterations in the reactive oxygen species (ROS) parameters in treated parasites were evident. Induction of apoptosis in treated parasites was confirmed by DNA laddering, acridine orange (AO)/ethidium bromide (EtBr) double staining and in situ DNA fragmentation. The downregulation of anti-apoptotic CED-9 and upregulation of proapoptotic EGL-1, CED-4 and CED-3 at both the transcription and translation levels confirmed apoptosis execution at the molecular level. Changes in the gene expressions of nuc-1, cps-6 and crn-1 further clarified the molecular cause of DNA degradation. Furthermore, azadirachtin was found to be non-toxic in both in vitro and in vivo toxicity analyses. Therefore, the experimental evidence detailed the pharmacological effectiveness of azadirachtin as a possible therapeutic agent against filariasis.

Keywords

anthelminticapoptosisazadirachtinlymphatic filariasisreactive oxygen speciesSetaria cervitoxicity
Type
Research Paper
Information
Journal of Helminthology , Volume 93 , Issue 5 , September 2019 , pp. 519 - 528
Copyright
Copyright © Cambridge University Press 2018 

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Footnotes

*

These authors contributed equally.

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