{"paper_id":"42f37968-e26e-4cb7-9670-06e021fea19d","body_text":"Lactate Alleviates Intestinal Barrier Injury in Weaned Piglets via Activation of the Wnt/β-Catenin Pathway and Promotion of Intestinal Epithelial Cell Proliferation1 | 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 Lactate Alleviates Intestinal Barrier Injury in Weaned Piglets via Activation of the Wnt/β-Catenin Pathway and Promotion of Intestinal Epithelial Cell Proliferation1 Mingyu Wang, Jiaojiao Chen, Aimin Wu, Daiwen Chen, Bing Yu, Jun He, and 6 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6878651/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 28 Nov, 2025 Read the published version in Journal of Animal Science and Biotechnology → Version 1 posted 4 You are reading this latest preprint version Abstract Background: Inflammatory bowel disease causes intestinal structural damage, impairs gut function, hinders animal growth and development, and reduces farming efficiency. Previous studies demonstrated that lactate alleviates Dextran Sulfate Sodium (DSS)-induced inflammation and mitigates weight loss by enhancing intestinal barrier functions. However, the mechanisms underlying lactate-mediated protection of the intestinal epithelial barrier unclear . This study aimed to explore the protective effect of lactate on intestinal barrier damage in colitis piglets and the possible underlying mechanisms through in vivo and in vitro experiments. Methods: A colitis model was established using weaned piglets administered DSS, with three experimental groups: control, DSS-treated, and DSS-treated with lactate supplementation. Jejunal morphology was assessed via Hematoxylin·and Eosin staining, while Western blotting quantified the expression levels of proliferation markers, including Cluster of Differentiation·24 (CD24) and Cyclin D1, and Wingless / Integrated (Wnt)/β-catenin signaling components. In vitro, DSS and sodium lactate treated Intestinal Porcine Epithelial Cell line-J2 (IPEC-J2) intestinal epithelial cells were assessed for viability (Cell Counting Kit-8 assay), apoptosis (flow cytometry), and proliferation parameters, including cell cycle analysis and Leucine-rich repeat-containing G-protein coupled receptor 5 (Lgr5+) stem cell quantification. Results: DSS administration induced significant jejunal villus shortening, downregulated CD24 and Cyclin D1 expression, and inhibited Wnt/β-catenin signaling in vivo. In vitro, DSS promoted apoptosis, inhibited proliferation, diminished the Lgr5+ cell populations, and reduced S-phase cell proportions. Conversely, lactate supplementation ameliorated DSS-induced villus atrophy, restored CD24 and Cyclin D1 levels, and activated Wnt/β-catenin signaling. Furthermore, in vitro, sodium lactate attenuated DSS-induced apoptosis, enhanced IPEC-J2 proliferation, expanded Lgr5+ cells, and increased S-phase progression. Conclusions: In summary, lactate ameliorated intestinal barrier damage in DSS-induced colitis by activating the Wnt/β-catenin pathway and restoring the balance between epithelial cell proliferation and apoptosis. This study provides novel mechanistic evidence supporting lactate’s therapeutic potential for IBD management. piglets lactate intestinal inflammation Wnt/β-catenin pathway poliferation and apoptosis Full Text Supplementary Files Supplementaryfigures.pdf supplementarymaterial.pdf Cite Share Download PDF Status: Published Journal Publication published 28 Nov, 2025 Read the published version in Journal of Animal Science and Biotechnology → Version 1 posted Reviewers agreed at journal 04 Jul, 2025 Reviewers invited by journal 17 Jun, 2025 Editor assigned by journal 13 Jun, 2025 First submitted to journal 12 Jun, 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-6878651\",\"acceptedTermsAndConditions\":true,\"allowDirectSubmit\":false,\"archivedVersions\":[],\"articleType\":\"Research Article\",\"associatedPublications\":[],\"authors\":[{\"id\":472437406,\"identity\":\"c2f3e16c-d43d-4c44-8415-647c673e4908\",\"order_by\":0,\"name\":\"Mingyu Wang\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Sichuan Agricultural University\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Mingyu\",\"middleName\":\"\",\"lastName\":\"Wang\",\"suffix\":\"\"},{\"id\":472437407,\"identity\":\"ab7a7cc7-8597-4912-969c-d3715ab0d412\",\"order_by\":1,\"name\":\"Jiaojiao 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Intestinal Epithelial Cell Proliferation1\",\"fulltext\":[],\"fulltextSource\":\"\",\"fullText\":\"\",\"funders\":[],\"hasAdminPriorityOnWorkflow\":false,\"hasManuscriptDocX\":false,\"hasOptedInToPreprint\":true,\"hasPassedJournalQc\":\"\",\"hasAnyPriority\":false,\"hideJournal\":false,\"highlight\":\"\",\"institution\":\"\",\"isAcceptedByJournal\":true,\"isAuthorSuppliedPdf\":true,\"isDeskRejected\":\"\",\"isHiddenFromSearch\":false,\"isInQc\":false,\"isInWorkflow\":false,\"isPdf\":true,\"isPdfUpToDate\":true,\"isWithdrawnOrRetracted\":false,\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"identity\":\"journal-of-animal-science-and-biotechnology\",\"isNatureJournal\":false,\"hasQc\":true,\"allowDirectSubmit\":false,\"externalIdentity\":\"jasb\",\"sideBox\":\"Learn more about [Journal of Animal Science and Biotechnology](http://jasbsci.biomedcentral.com/)\",\"snPcode\":\"\",\"submissionUrl\":\"https://www.editorialmanager.com/jasb/default.aspx\",\"title\":\"Journal of Animal Science and Biotechnology\",\"twitterHandle\":\"@animalplantsci\",\"acdcEnabled\":true,\"dfaEnabled\":true,\"editorialSystem\":\"em\",\"reportingPortfolio\":\"BMC/SO AJ\",\"inReviewEnabled\":true,\"inReviewRevisionsEnabled\":true},\"keywords\":\"piglets, lactate, intestinal inflammation, Wnt/β-catenin pathway, poliferation and apoptosis\",\"lastPublishedDoi\":\"10.21203/rs.3.rs-6878651/v1\",\"lastPublishedDoiUrl\":\"https://doi.org/10.21203/rs.3.rs-6878651/v1\",\"license\":{\"name\":\"CC BY 4.0\",\"url\":\"https://creativecommons.org/licenses/by/4.0/\"},\"manuscriptAbstract\":\"\\u003cp\\u003e\\u003cstrong\\u003eBackground:\\u003c/strong\\u003e Inflammatory bowel disease causes intestinal structural damage, impairs gut function, hinders animal growth and development, and reduces farming efficiency. Previous studies demonstrated that lactate alleviates Dextran Sulfate Sodium (DSS)-induced inflammation and mitigates weight loss by enhancing intestinal barrier functions. However,\\u003cbr\\u003e\\nthe mechanisms underlying lactate-mediated protection of the intestinal epithelial barrier unclear . This study aimed to explore the protective effect of lactate on intestinal barrier damage in colitis piglets and the possible underlying mechanisms through in vivo and in vitro experiments.\\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eMethods: \\u003c/strong\\u003eA colitis model was established using weaned piglets administered DSS, with three experimental groups: control, DSS-treated, and DSS-treated with lactate supplementation. Jejunal morphology was assessed via Hematoxylin·and Eosin staining, while Western blotting quantified the expression levels of proliferation markers, including Cluster of Differentiation·24 (CD24) and Cyclin D1, and Wingless / Integrated (Wnt)/β-catenin signaling components. In vitro, DSS and sodium lactate treated Intestinal Porcine Epithelial Cell line-J2 (IPEC-J2) intestinal epithelial cells were assessed for viability (Cell Counting Kit-8 assay), apoptosis (flow cytometry), and proliferation parameters, including cell cycle analysis and Leucine-rich repeat-containing G-protein coupled receptor 5 (Lgr5+) stem cell quantification.\\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eResults:\\u003c/strong\\u003e DSS administration induced significant jejunal villus shortening, downregulated CD24 and Cyclin D1 expression, and inhibited Wnt/β-catenin signaling in vivo. In vitro, DSS promoted apoptosis, inhibited proliferation, diminished the Lgr5+ cell populations, and reduced S-phase cell proportions. Conversely, lactate supplementation ameliorated DSS-induced villus atrophy, restored CD24 and Cyclin D1 levels, and activated Wnt/β-catenin signaling. Furthermore, in vitro, sodium lactate attenuated DSS-induced apoptosis, enhanced IPEC-J2 proliferation, expanded Lgr5+ cells, and increased S-phase progression.\\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eConclusions:\\u003c/strong\\u003e In summary, lactate ameliorated intestinal barrier damage in DSS-induced colitis by activating the Wnt/β-catenin pathway and restoring the balance between epithelial cell proliferation and apoptosis. This study provides novel mechanistic evidence supporting lactate’s therapeutic potential for IBD management.\\u003c/p\\u003e\",\"manuscriptTitle\":\"Lactate Alleviates Intestinal Barrier Injury in Weaned Piglets via Activation of the Wnt/β-Catenin Pathway and Promotion of Intestinal Epithelial Cell Proliferation1\",\"msid\":\"\",\"msnumber\":\"\",\"nonDraftVersions\":[{\"code\":1,\"date\":\"2025-06-19 05:37:08\",\"doi\":\"10.21203/rs.3.rs-6878651/v1\",\"editorialEvents\":[{\"type\":\"communityComments\",\"content\":0},{\"type\":\"reviewerAgreed\",\"content\":\"\",\"date\":\"2025-07-04T06:54:15+00:00\",\"index\":0,\"fulltext\":\"\"},{\"type\":\"reviewersInvited\",\"content\":\"\",\"date\":\"2025-06-17T08:55:15+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"editorAssigned\",\"content\":\"\",\"date\":\"2025-06-13T13:45:41+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"submitted\",\"content\":\"Journal of Animal Science and Biotechnology\",\"date\":\"2025-06-12T22:21:16+00:00\",\"index\":\"\",\"fulltext\":\"\"}],\"status\":\"published\",\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"identity\":\"journal-of-animal-science-and-biotechnology\",\"isNatureJournal\":false,\"hasQc\":true,\"allowDirectSubmit\":false,\"externalIdentity\":\"jasb\",\"sideBox\":\"Learn more about [Journal of Animal Science and Biotechnology](http://jasbsci.biomedcentral.com/)\",\"snPcode\":\"\",\"submissionUrl\":\"https://www.editorialmanager.com/jasb/default.aspx\",\"title\":\"Journal of Animal Science and Biotechnology\",\"twitterHandle\":\"@animalplantsci\",\"acdcEnabled\":true,\"dfaEnabled\":true,\"editorialSystem\":\"em\",\"reportingPortfolio\":\"BMC/SO AJ\",\"inReviewEnabled\":true,\"inReviewRevisionsEnabled\":true}}],\"origin\":\"\",\"ownerIdentity\":\"fe73eff0-74af-467d-b75a-745d3c726566\",\"owner\":[],\"postedDate\":\"June 19th, 2025\",\"published\":true,\"recentEditorialEvents\":[],\"rejectedJournal\":[],\"revision\":\"\",\"amendment\":\"\",\"status\":\"published-in-journal\",\"subjectAreas\":[],\"tags\":[],\"updatedAt\":\"2025-12-01T16:01:07+00:00\",\"versionOfRecord\":{\"articleIdentity\":\"rs-6878651\",\"link\":\"https://doi.org/10.1186/s40104-025-01290-x\",\"journal\":{\"identity\":\"journal-of-animal-science-and-biotechnology\",\"isVorOnly\":false,\"title\":\"Journal of Animal Science and Biotechnology\"},\"publishedOn\":\"2025-11-28 15:57:27\",\"publishedOnDateReadable\":\"November 28th, 2025\"},\"versionCreatedAt\":\"2025-06-19 05:37:08\",\"video\":\"\",\"vorDoi\":\"10.1186/s40104-025-01290-x\",\"vorDoiUrl\":\"https://doi.org/10.1186/s40104-025-01290-x\",\"workflowStages\":[]},\"version\":\"v1\",\"identity\":\"rs-6878651\",\"journalConfig\":\"researchsquare\"},\"__N_SSP\":true},\"page\":\"/article/[identity]/[[...version]]\",\"query\":{\"redirect\":\"/article/rs-6878651\",\"identity\":\"rs-6878651\",\"version\":[\"v1\"]},\"buildId\":\"8U1c8b4HqxoKbykW_rLl7\",\"isFallback\":false,\"isExperimentalCompile\":false,\"dynamicIds\":[84888],\"gssp\":true,\"scriptLoader\":[]}","source_license":"CC-BY-4.0","license_restricted":false}