Polyinosinic polycytidylic acid (poly I:C) Induces Neuronal Cell Death Through NLRP3-mediated inflammasome in human microglia and neuroinflammation-induced cognitive impairment in mice

Polyinosinic polycytidylic acid (poly I:C) Induces Neuronal Cell Death Through NLRP3-mediated inflammasome in human microglia and neuroinflammation-induced cognitive impairment in mice | 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 Polyinosinic polycytidylic acid (poly I:C) Induces Neuronal Cell Death Through NLRP3-mediated inflammasome in human microglia and neuroinflammation-induced cognitive impairment in mice Aseel Y Altahrawi, Antonisamy William James, Zahoor A. Shah This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7272392/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 19 Nov, 2025 Read the published version in Molecular Neurobiology → Version 1 posted 10 You are reading this latest preprint version Abstract Neuroinflammation and neuronal death are direct consequences of persistent microglial activation observed in many chronic neurological conditions. Activated microglia impact neuronal cells by releasing proinflammatory cytokines and inflammatory mediators, leading to neuronal damage and neurodegeneration. To investigate whether Polyinosinic polycytidylic acid (poly I:C), a synthetic double-stranded RNA molecule, induces neuroinflammation and neuronal death, we exposed microglia (HMC-3 cells) to poly I: C for 24 hrs, and assessed inflammatory cytokines. Additionally, to investigate whether poly(I:C) induces memory impairment and motor coordination deficits in C57BL/6 mice, we conducted a behavioural assessment and measured the expression of inflammatory cytokines in the brain. Poly (I:C) exposure significantly increased the mRNA and protein expression of inflammasome, proinflammatory cytokines (TNFα, IL-6, IL-1β, IL-8, IL-12, and IL-18) and chemokines in microglia. Poly (I: C) also significantly increased the translocation of NF-kB from the cytosol to the nucleus. Furthermore, the conditioned medium from poly(I:C)-treated cells markedly increased apoptosis in human neuronal cells (differentiated SHSY5Y cells) by activating pro-apoptotic markers, including Bax, Bad, cleaved caspase-3, cleaved PARP, and AIF. In mice, exposure to poly (I:C) significantly increased the mRNA expression of inflammatory cytokines, such as IL-6 and TNF-α, in the hippocampus. It also decreased the percentage of alternation in the T-maze test and reduced the distance travelled and average speed in the open field test, indicating motor and cognitive deficits. These findings suggest that poly (I:C) induces neuroinflammation through the inflammasome and proinflammatory mediators via the NLRP3/NF-κB signaling pathway in vitro and in vivo and induces motor and memory impairment in mice. Poly (I C) neuroinflammation proinflammatory cytokines neuronal death memory impairment Full Text Additional Declarations No competing interests reported. Supplementary Files Supplementalfile.docx Cite Share Download PDF Status: Published Journal Publication published 19 Nov, 2025 Read the published version in Molecular Neurobiology → Version 1 posted Editorial decision: Revision requested 12 Sep, 2025 Reviews received at journal 26 Aug, 2025 Reviews received at journal 25 Aug, 2025 Reviewers agreed at journal 20 Aug, 2025 Reviewers agreed at journal 20 Aug, 2025 Reviewers agreed at journal 18 Aug, 2025 Reviewers invited by journal 18 Aug, 2025 Submission checks completed at journal 12 Aug, 2025 Editor assigned by journal 12 Aug, 2025 First submitted to journal 01 Aug, 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. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. 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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-7272392","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":505416281,"identity":"ccd294de-6a70-42c0-8493-af00e25aaa57","order_by":0,"name":"Aseel Y Altahrawi","email":"","orcid":"","institution":"The University of Toledo","correspondingAuthor":false,"prefix":"","firstName":"Aseel","middleName":"Y","lastName":"Altahrawi","suffix":""},{"id":505416283,"identity":"62f5ef41-b228-47ed-a96e-96ed1d6320e8","order_by":1,"name":"Antonisamy William James","email":"","orcid":"","institution":"The University of Toledo","correspondingAuthor":false,"prefix":"","firstName":"Antonisamy","middleName":"William","lastName":"James","suffix":""},{"id":505416284,"identity":"dc605ce4-5dc0-4fb0-9845-c54dfede26df","order_by":2,"name":"Zahoor A. 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Activated microglia impact neuronal cells by releasing proinflammatory cytokines and inflammatory mediators, leading to neuronal damage and neurodegeneration. To investigate whether Polyinosinic polycytidylic acid (poly I:C), a synthetic double-stranded RNA molecule, induces neuroinflammation and neuronal death, we exposed microglia (HMC-3 cells) to poly I: C for 24 hrs, and assessed inflammatory cytokines. Additionally, to investigate whether poly(I:C) induces memory impairment and motor coordination deficits in C57BL/6 mice, we conducted a behavioural assessment and measured the expression of inflammatory cytokines in the brain. Poly (I:C) exposure significantly increased the mRNA and protein expression of inflammasome, proinflammatory cytokines (TNFα, IL-6, IL-1β, IL-8, IL-12, and IL-18) and chemokines in microglia. Poly (I: C) also significantly increased the translocation of NF-kB from the cytosol to the nucleus. 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