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Viral inhibitory potential of hyoscyamine in Japanese encephalitis virus–infected embryonated chicken eggs involving multiple signaling pathways

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Abstract

Japanese encephalitis virus (JEV) is the leading cause of viral encephalitis worldwide. The emergence of new genotypes of the virus and a high rate of mutation make it necessary to develop alternative treatment strategies against this deadly pathogen. Although the antiviral properties of Atropa belladonna and some of its active components, such as atropine and scopolamine, have been studied, the effect of another important component, hyoscyamine, against JEV infection has not yet been investigated. In this study, we investigated the antiviral effect of hyoscyamine against JEV and its immunomodulatory activity in embryonated chicken eggs. Pretreatment with hyoscyamine sulphate resulted in a significant decrease in the viral load in both chorioallantoic membrane (CAM) and brain tissues at 48 and 96 hours postinfection. In silico studies showed stable binding and interaction between hyoscyamine and non-structural protein 5 (NS5), suggesting that this could be the basis of its antiviral effect. Embryonated eggs pretreated with hyoscyamine sulphate showed upregulation of Toll-like receptor 3 (TLR3), TLR7, TLR8, interleukin 4 (IL-4), and IL-10 as well as interferons and regulatory factors. Hyoscyamine sulphate was also found to cause significant downregulation of TLR4. The potential use of hyoscyamine for controlling JEV replication and its dissemination to the brain suggest that it may be a promising therapy option against JEV in the future.

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Abbreviations

JEV:

Japanese encephalitis virus

IFN:

Interferon

RH:

Relative humidity

BSL:

Biosafety level

TLR:

Toll-like receptor

RIG:

Retinoic acid-inducible gene

MDA:

Melanoma differentiation associated gene

PAMP:

Pathogen-associated molecular pattern

ERK:

Extracellular signal-regulated kinase

MAPK:

Mitogen-activated protein kinase

AP:

Activator protein

CAM:

Chorioallantoic membrane

BBB:

Blood brain barrier

NIV:

National Institute of Virology

PBS:

Phosphate-buffered saline

BSA:

Bovine serum albumin

RCSB:

Research collaborator for structural bioinformatics

PDB:

Protein Data Bank

PDBQT:

Protein Data Bank, Partial Charge (Q), and Atom Type (T)

MD:

Molecular dynamics

RMSD:

Root mean square deviation

RMSF:

Root mean square fluctuation

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

ANOVA:

Analysis of variance

NS:

Non-structural

IRF:

Interferon regulatory factor

IL:

Interleukin

TRIF:

TIR-domain-containing adaptor-inducing interferon β

NF-κB:

Nuclear factor kappa light chain enhancer of activated B cells

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Acknowledgements

The authors are extremely thankful to the Department of Biotechnology, St. Xavier’s College, Kolkata, Department of Cardiology, R.G Kar Medical College & Hospital, Sarada Multispeciality Hospital, for providing us with different facilities for the wet lab experiments. We also acknowledge Om Saswat Sahoo from the Department of Biotechnology, National Institute of Technology Duragpur for his sincere help and contributions to this article. We are also deeply indebted to Rev. Dr. Dominic Savio, S.J, Principal & Rector, St. Xavier’s College, Kolkata, Dr. Satadal Das, Consultant Senior Microbiologist, Peerless Hospital & B.K Roy Research Centre. We also convey our gratitude to the National Institute of Virology (NIV) Pune for providing us with the viral strains.

Funding

This work was not supported by any funding agency.

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Authors and Affiliations

  1. Department of Biotechnology, National Institute of Technology, Durgapur, West Bengal, India

    Arghyadeep Bhattacharjee & Rahul Naga

  2. Department of Microbiology, Kingston College of Science, Beruanpukuria, Malikapur, Kolkata-126, West Bengal, India

    Arghyadeep Bhattacharjee & Srabani Karmakar

  3. Department of Cardiology, R.G Kar Medical College and Hospital, Kolkata, West Bengal, India

    Manish Saha

  4. Department of Biotechnology, St. Xavier’s College, Kolkata, West Bengal, India

    Souvik Roy

  5. Department of Microbiology, Ramkrishna Mission Vidyamandira, Belur, Howrah, West Bengal, India

    Abhishek Pal

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Contributions

Study conception and design: AB, MS. Acquisition of data: AB, RN, MS, AP, SK. Analysis and interpretation of data: AB, RN. Drafting of manuscript: AB. Critical revision: AB, MS, AP. Supervision and research resources supply: RN, MS, SK, SR. All authors read and approved the final version of the manuscript.

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Correspondence to Arghyadeep Bhattacharjee.

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Bhattacharjee, A., Naga, R., Saha, M. et al. Viral inhibitory potential of hyoscyamine in Japanese encephalitis virus–infected embryonated chicken eggs involving multiple signaling pathways. Arch Virol 168, 264 (2023). https://doi.org/10.1007/s00705-023-05883-7

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