Scientific Validation of the Traditional Anti-Inflammatory Use of the Sapindus mukorossi Seed Kernel: In Vitro and In Silico Insights

Scientific Validation of the Traditional Anti-Inflammatory Use of the Sapindus mukorossi Seed Kernel: In Vitro and In Silico Insights | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Scientific Validation of the Traditional Anti-Inflammatory Use of the Sapindus mukorossi Seed Kernel: In Vitro and In Silico Insights Esther Lathazuali, Hmingremhlua Sailo, Martin Lalnunthara, No given name Lalremliani, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8101955/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 18 You are reading this latest preprint version Abstract S. mukorossi ., commonly known as soapnut, is a traditional medicinal plant widely used in Asian ethnomedicine for treating infections and inflammatory disorders. However, systematic validation of its pharmacological potential remains limited. The antimicrobial and anti-inflammatory properties of S. mukorossi extract were evaluated using in vitro and biochemical assays. Cytotoxicity was assessed in RAW 264.7 macrophages using the MTT assay. Antibacterial activity was determined by agar well diffusion and minimum inhibitory concentration (MIC) assays against Escherichia coli, Klebsiella pneumoniae , and Staphylococcus aureus , while antifungal efficacy was tested against Candida albicans using disc diffusion. Anti-inflammatory potential was examined by measuring TNF-α levels through ELISA and cyclooxygenase-2 (COX-2) inhibition via a colorimetric assay. Molecular docking and molecular dynamics simulations, along with gmx_MMPBSA free energy calculations, were performed to assess the binding affinity of identified compounds (isosilybin A, orientin) with IRAK4. The extract demonstrated significant antimicrobial activity, producing notable inhibition zones against both Gram-positive and Gram-negative bacteria, with low MIC values indicating potent bacteriostatic effects. Antifungal activity against C. albicans was comparable to amphotericin B. The extract exhibited low cytotoxicity toward RAW 264.7 cells and effectively suppressed inflammatory mediators, showing dose-dependent inhibition of COX-2 activity and a marked reduction in TNF-α secretion. In silico analyses revealed stable binding of isosilybin A and orientin to IRAK4, as confirmed by molecular dynamics and gmx_MMPBSA free energy calculations (ΔGbind up to − 47.8 kcal/mol). The findings substantiate the traditional use of S. mukorossi as a natural antimicrobial and anti-inflammatory agent. Its bioactive constituents may serve as potential leads for developing plant-derived therapeutic agents targeting infectious and inflammatory diseases. Further phytochemical characterization and molecular mechanism studies are warranted to elucidate the active principles and their targets. Sapindus mukorossi Anti-inflammatory activity Haemorrhoids Molecular docking and dynamics Ethnopharmacology Full Text Additional Declarations No competing interests reported. Supplementary Files suppl..docx Cite Share Download PDF Status: Under Review Version 1 posted Editorial decision: Revision requested 16 Feb, 2026 Reviews received at journal 11 Feb, 2026 Reviews received at journal 28 Jan, 2026 Reviews received at journal 28 Jan, 2026 Reviewers agreed at journal 26 Jan, 2026 Reviews received at journal 25 Jan, 2026 Reviewers agreed at journal 25 Jan, 2026 Reviewers agreed at journal 25 Jan, 2026 Reviewers agreed at journal 25 Jan, 2026 Reviewers agreed at journal 24 Jan, 2026 Reviewers agreed at journal 23 Jan, 2026 Reviewers agreed at journal 23 Jan, 2026 Reviewers agreed at journal 23 Jan, 2026 Reviewers invited by journal 23 Jan, 2026 Editor invited by journal 12 Jan, 2026 Editor assigned by journal 12 Jan, 2026 Submission checks completed at journal 18 Dec, 2025 First submitted to journal 18 Dec, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-8101955","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":580460026,"identity":"47f3c723-4226-4cc6-9cd6-1caa3accbd12","order_by":0,"name":"Esther Lathazuali","email":"","orcid":"","institution":"Mizoram University","correspondingAuthor":false,"prefix":"","firstName":"Esther","middleName":"","lastName":"Lathazuali","suffix":""},{"id":580460031,"identity":"eddc9594-0033-4117-ab96-3c77f55bd161","order_by":1,"name":"Hmingremhlua Sailo","email":"","orcid":"","institution":"Pachhunga University 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However, systematic validation of its pharmacological potential remains limited. The antimicrobial and anti-inflammatory properties of \u003cem\u003eS. mukorossi\u003c/em\u003e extract were evaluated using in vitro and biochemical assays. Cytotoxicity was assessed in RAW 264.7 macrophages using the MTT assay. Antibacterial activity was determined by agar well diffusion and minimum inhibitory concentration (MIC) assays against \u003cem\u003eEscherichia coli, Klebsiella pneumoniae\u003c/em\u003e, and \u003cem\u003eStaphylococcus aureus\u003c/em\u003e, while antifungal efficacy was tested against \u003cem\u003eCandida albicans\u003c/em\u003e using disc diffusion. Anti-inflammatory potential was examined by measuring TNF-α levels through ELISA and cyclooxygenase-2 (COX-2) inhibition via a colorimetric assay. 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