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Japanese encephalitis virus (JEV) is the leading cause of viral encephalitis worldwide. Emergence of new genotypes of the virus and increased rate of mutation demands for alternatives that can fight against this deadly pathogen. Although the anti-viral property of Atropa belladonna and its active components such as Atropine, Scopolamine has been studied, yet the role of its another important component hyoscyamine against JEV has not yet been established till date. Thus our current study is focussed to investigate the protective effects of hyoscyamine against JEV and its immunomodulatory function in embryonated chick. Pre-treatment with hyoscyamine sulphate indicated significant decrease in viral load in both CAM and brain tissues after 48 hour and 96 hour (post infection). Insilico studies showed stable binding and interaction between hyoscyamine and NS5 (non structural protein 5) making it an effective compound against the virus. Embryonated eggs pre-treated with hyoscyamine sulphate showed upregulation of TLR3, TLR7, TLR8, IL-4, IL-10 as well as interferons and the regulatory factors. Hyoscyamine sulphate caused significant downregulation of TLR4 as compared to other experimental groups as analysed by one-way ANOVA. Hence our study demonstrated the protective function of hyoscyamine in controlling JEV viral replication and its dissemination to brain that may emerge as a promising therapy against JEV in future.

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