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Through molecular docking, this study revealed that mangiferin has a strong binding affinity for the active site of the neuraminidase (NA) protein of influenza virus A(H1N1)pdm09, with a binding energy of \u22128.1 kcal\/mol. In vitro assays confirmed a dose-dependent inhibition of NA, with an IC50 of 88.65 \u03bcM, and minimal cytotoxicity, as indicated by a CC50 of 328.1 \u03bcM in MDCK cells. In murine models, the administration of mangiferin at a dosage of 25 mg\/kg significantly mitigated weight loss, decreased viral loads in nasal turbinates and lungs by over 1 log10 TCID50, and enhanced survival rates from 0% in control groups to 20% in mangiferin-treated group at 14 days post-infection. In addition, mangiferin was found to modulate host immune responses by simultaneously inhibiting pro-inflammatory cytokines, IL-6 and TNF-\u03b1, and upregulating the expression of anti-inflammatory IL-10 and antiviral IFN-\u03b3, thus mitigating infection-induced inflammation. Our findings elucidate the dual mechanism of mangiferin involving the direct inhibition of NA and immunomodulation, thereby providing experimental evidence for exploring dual-mechanism-based anti-influenza strategies against resistant strains of influenza.<\/jats:p>","DOI":"10.3390\/v17070873","type":"journal-article","created":{"date-parts":[[2025,6,24]],"date-time":"2025-06-24T10:44:41Z","timestamp":1750761881000},"page":"873","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Integrated In Silico, In Vitro, and In Vivo Studies Reveal Mangiferin as a Promising Antiviral Agent Against H1N1\/pdm2009 Influenza Virus"],"prefix":"10.3390","volume":"17","author":[{"given":"Yinde","family":"Gan","sequence":"first","affiliation":[{"name":"State Key Laboratory for Animal Disease Control and Prevention, Center for Emerging and Zoonotic Diseases, College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fucheng","family":"Guo","sequence":"additional","affiliation":[{"name":"State Key Laboratory for Animal Disease Control and Prevention, Center for Emerging and Zoonotic Diseases, College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China"},{"name":"College of Coastal Agricultural Science, Guangdong Ocean University, Zhanjiang 524088, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ayan","family":"Roy","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, Asian University for Women, Chittagong 4000, Bangladesh"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiao","family":"Wang","sequence":"additional","affiliation":[{"name":"State Key Laboratory for Animal Disease Control and Prevention, Center for Emerging and Zoonotic Diseases, College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7660-5522","authenticated-orcid":false,"given":"Yongyi","family":"Shen","sequence":"additional","affiliation":[{"name":"State Key Laboratory for Animal Disease Control and Prevention, Center for Emerging and Zoonotic Diseases, College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China"},{"name":"School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,6,21]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1529","DOI":"10.1126\/science.1189132","article-title":"Reassortment of pandemic H1N1\/2009 influenza A virus in swine","volume":"328","author":"Vijaykrishna","year":"2010","journal-title":"Science"},{"key":"ref_2","first-page":"415","article-title":"The structural variability of the influenza A hemagglutinin receptor-binding site","volume":"17","author":"Lazniewski","year":"2018","journal-title":"Brief. 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