Extracellular vesicles containing anti-miR-128-3p derived from mesenchymal stem cells attenuate cerebral ischemia-reperfusion injury by suppressing neuronal pyroptosis and modulating microglia polarization

Extracellular vesicles containing anti-miR-128-3p derived from mesenchymal stem cells attenuate cerebral ischemia-reperfusion injury by suppressing neuronal pyroptosis and modulating microglia polarization | 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 Extracellular vesicles containing anti-miR-128-3p derived from mesenchymal stem cells attenuate cerebral ischemia-reperfusion injury by suppressing neuronal pyroptosis and modulating microglia polarization Xinyu Zhou, Hang Zhou, Liaoyang Xu, Huijuan Wu, Xiaomei Zhou, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-3964083/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Cerebral ischemia-reperfusion (CI/R) injury seriously threatens the patients with ischemic stroke. MiR-128-3p level was found to be upregulated in serum samples of patients with ischemic stroke. It has been shown that miRNA in extracellular vesicles (specifically exosomes) has attracted attention because mesenchymal stem cells (MSCs)-derived exosomes play a favorable role in improving CI/R injury. Thus, we investigated whether MSCs-derived exosomes can be used to deliver miR-128-3p inhibitor (anti-miR-128-3p) to treat CI/R injury. BV2 cells were exposed to oxygen-glucose deprivation/reperfusion (OGD/R) for constructing in vitro model of CI/R. In addition, a rat model of middle cerebral artery occlusion (MCAO) was established to construct in vivo model of CI/R. Results indicated that miR-128-3p inhibitor can be transferred from MSCs to BV2 cells via exosomes. MSCs-derived exosomes containing anti-miR-128-3p significantly reduced the viability and inflammatory response in OGD/R-treated BV2 cells. Additionally, exosomes containing anti-miR-128-3p was able to enhance M1-to-M2 polarization of BV2 cells exposed to OGD/R. Meanwhile, exosomal anti-miR-128-3p markedly decreased infarct area in MCAO rats. Furthermore, NRBF2 was a direct target of miR-128-3p. Exosomal anti-miR-128-3p obviously reduced the level of NRBF2 downstream genes including NLRP3 and Caspase 1 in OGD/R-treated BV2 cells as well as in brain tissues of MCAO rats, suggesting that MSCs-derived exosomal anti-miR-128-3p could attenuate neural cell pyroptosis. In conclusion, exosomes containing anti-miR-128-3p derived from MSCs could attenuate CI/R injury via suppressing neuronal pyroptosis and modulating microglia polarization. Hence, our study might provide a theoretical basis for the treatment of ischemic stroke. Ischemic stroke cerebral ischemia-reperfusion injury microRNA microglial M1 polarization cell pyroptosis Full Text Additional Declarations No competing interests reported. Supplementary Files SupplementarymaterialCPM131Procell2.pdf Cite Share Download PDF Status: Posted Version 1 posted 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. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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-3964083","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":273923390,"identity":"b2c2ed55-3945-43ef-993d-1a6a7b1ffb82","order_by":0,"name":"Xinyu Zhou","email":"","orcid":"","institution":"Lianyungang Clinical College of Nanjing Medical University","correspondingAuthor":false,"prefix":"","firstName":"Xinyu","middleName":"","lastName":"Zhou","suffix":""},{"id":273923391,"identity":"5b31a65f-4531-4260-9b72-e468e549a001","order_by":1,"name":"Hang Zhou","email":"","orcid":"","institution":"The Affiliated Lianyungang Hospital of Xuzhou Medical University","correspondingAuthor":false,"prefix":"","firstName":"Hang","middleName":"","lastName":"Zhou","suffix":""},{"id":273923392,"identity":"24c30c1e-66cd-474d-be51-9671513edc50","order_by":2,"name":"Liaoyang Xu","email":"","orcid":"","institution":"The Affiliated Lianyungang Hospital of Xuzhou Medical University","correspondingAuthor":false,"prefix":"","firstName":"Liaoyang","middleName":"","lastName":"Xu","suffix":""},{"id":273923393,"identity":"e3735b4c-8371-4418-b788-7cdc56336b36","order_by":3,"name":"Huijuan Wu","email":"","orcid":"","institution":"Jinzhou Medical University","correspondingAuthor":false,"prefix":"","firstName":"Huijuan","middleName":"","lastName":"Wu","suffix":""},{"id":273923394,"identity":"74a6e605-4899-426b-99e4-c1dcee6f36c3","order_by":4,"name":"Xiaomei Zhou","email":"","orcid":"","institution":"The Affiliated Lianyungang Hospital of Xuzhou Medical University","correspondingAuthor":false,"prefix":"","firstName":"Xiaomei","middleName":"","lastName":"Zhou","suffix":""},{"id":273923395,"identity":"240b2926-9e32-463e-94dd-61226b158e2f","order_by":5,"name":"Xiaolin Ren","email":"","orcid":"","institution":"The First Affiliated Hospital of Kangda College of Nanjing Medical University","correspondingAuthor":false,"prefix":"","firstName":"Xiaolin","middleName":"","lastName":"Ren","suffix":""},{"id":273923396,"identity":"2e3deb7e-97a8-4da5-9060-51b2d3a2e681","order_by":6,"name":"Xiaomin Li","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAzUlEQVRIiWNgGAWjYBACPgYGxgMMDDYMjA0gLhsRWkBqgFrSSNdyGIlLUItE+oXDvDnn85innTFg+FB2mIF/dgMhLTkFB2duu13MODvHgHHGucMMEncOENSScODjttuJjUAtzLxthxkMJBKI0JK47RxEy1/itKQfANpyAKKFkSgtPG8YgH5JBmpJKzjYcy6dR+IGAS387OkPH/Nus0vcODt544MfZdZy/DMIaGEQyDEA04YN4Ahi4CGgHmTN8QdgWp6w0lEwCkbBKBipAADjiEejT0pwpAAAAABJRU5ErkJggg==","orcid":"","institution":"Lianyungang Clinical College of Nanjing Medical University","correspondingAuthor":true,"prefix":"","firstName":"Xiaomin","middleName":"","lastName":"Li","suffix":""}],"badges":[],"createdAt":"2024-02-17 12:45:56","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-3964083/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-3964083/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":53179277,"identity":"3ef6a7f3-fc95-455c-af04-55b3e86e8f32","added_by":"auto","created_at":"2024-03-21 15:21:32","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":718389,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3964083/v1_covered_60917d56-3551-4e78-8333-70c400b9bf49.pdf"},{"id":51436749,"identity":"8ae68e20-86c9-4bcb-a1df-96b6d28f99f8","added_by":"auto","created_at":"2024-02-21 15:30:46","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":552415,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementarymaterialCPM131Procell2.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3964083/v1/c8b21edba27f3b4c8642edb4.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Extracellular vesicles containing anti-miR-128-3p derived from mesenchymal stem cells attenuate cerebral ischemia-reperfusion injury by suppressing neuronal pyroptosis and modulating microglia polarization","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Ischemic stroke, cerebral ischemia-reperfusion injury, microRNA, microglial M1 polarization, cell pyroptosis","lastPublishedDoi":"10.21203/rs.3.rs-3964083/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-3964083/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eCerebral ischemia-reperfusion (CI/R) injury seriously threatens the patients with ischemic stroke. MiR-128-3p level was found to be upregulated in serum samples of patients with ischemic stroke. It has been shown that miRNA in extracellular vesicles (specifically exosomes) has attracted attention because mesenchymal stem cells (MSCs)-derived exosomes play a favorable role in improving CI/R injury. Thus, we investigated whether MSCs-derived exosomes can be used to deliver miR-128-3p inhibitor (anti-miR-128-3p) to treat CI/R injury. BV2 cells were exposed to oxygen-glucose deprivation/reperfusion (OGD/R) for constructing \u003cem\u003ein vitro\u003c/em\u003e model of CI/R. In addition, a rat model of middle cerebral artery occlusion (MCAO) was established to construct \u003cem\u003ein vivo\u003c/em\u003e model of CI/R. Results indicated that miR-128-3p inhibitor can be transferred from MSCs to BV2 cells via exosomes. MSCs-derived exosomes containing anti-miR-128-3p significantly reduced the viability and inflammatory response in OGD/R-treated BV2 cells. Additionally, exosomes containing anti-miR-128-3p was able to enhance M1-to-M2 polarization of BV2 cells exposed to OGD/R. Meanwhile, exosomal anti-miR-128-3p markedly decreased infarct area in MCAO rats. Furthermore, NRBF2 was a direct target of miR-128-3p. Exosomal anti-miR-128-3p obviously reduced the level of NRBF2 downstream genes including NLRP3 and Caspase 1 in OGD/R-treated BV2 cells as well as in brain tissues of MCAO rats, suggesting that MSCs-derived exosomal anti-miR-128-3p could attenuate neural cell pyroptosis. In conclusion, exosomes containing anti-miR-128-3p derived from MSCs could attenuate CI/R injury via suppressing neuronal pyroptosis and modulating microglia polarization. Hence, our study might provide a theoretical basis for the treatment of ischemic stroke.\u003c/p\u003e","manuscriptTitle":"Extracellular vesicles containing anti-miR-128-3p derived from mesenchymal stem cells attenuate cerebral ischemia-reperfusion injury by suppressing neuronal pyroptosis and modulating microglia polarization","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-02-21 15:30:41","doi":"10.21203/rs.3.rs-3964083/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"7a8b63c4-21fd-4660-9a65-195db1b55266","owner":[],"postedDate":"February 21st, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-03-21T15:13:06+00:00","versionOfRecord":[],"versionCreatedAt":"2024-02-21 15:30:41","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-3964083","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-3964083","identity":"rs-3964083","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}