Tubuloside B Ameliorates Blood-Brain Barrier Disruption and Cognitive Deficits after Cerebral Ischemia by Inhibiting TRIC Protein Ubiquitination and Degradation

Tubuloside B Ameliorates Blood-Brain Barrier Disruption and Cognitive Deficits after Cerebral Ischemia by Inhibiting TRIC Protein Ubiquitination and Degradation | 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 Tubuloside B Ameliorates Blood-Brain Barrier Disruption and Cognitive Deficits after Cerebral Ischemia by Inhibiting TRIC Protein Ubiquitination and Degradation Mingli He, Han Zhang, Ru Liu, Chunyang Zhang, Juan Li, Luming Li, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7553882/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 29 Dec, 2025 Read the published version in Journal of Neuroinflammation → Version 1 posted 11 You are reading this latest preprint version Abstract Background and Purpose Vascular cognitive impairment (VCI) is strongly linked to blood-brain barrier (BBB) dysfunction. This study investigated whether Tubuloside B (Tub-B), a primary bioactive compound of Cistanche tubulosa, ameliorates cognitive deficits by protecting BBB integrity through inhibition of tricellulin (TRIC) ubiquitination and degradation. Methods The main plasma metabolites of total glycosides of Cistanche (TCGs) were identified by UPLC-QQQ-MS. A bilateral common carotid artery occlusion (BCCAO) rat model was used to induce chronic cerebral hypoperfusion (CCH). Cognitive function was assessed using the Morris water maze, and BBB permeability was evaluated by Evans blue extravasation. TRIC expression and ubiquitination were analyzed by Western blot, co-immunoprecipitation, and immunohistochemistry. An in vitro oxygen-glucose deprivation (OGD) model in brain microvascular endothelial cells was used to validate the effects on barrier function. Results Tub-B was identified as the major circulating metabolite, accounting for 52.3% ± 2.1% of plasma components. In CCH rats, Tub-B (20 mg/kg) significantly improved spatial memory, increasing platform crossings (3.00 ± 0.58 vs. CCH, P < 0.05) and reducing Evans blue leakage (0.038 ± 0.007 µg/g tissue vs. 0.15 ± 0.02 µg/g in CCH, P < 0.001). It also attenuated the downregulation of TRIC and suppressed its ubiquitination. In OGD-injured endothelial cells, Tub-B enhanced barrier integrity and inhibited TRIC ubiquitination. Conclusion Tub-B attenuates CCH-induced BBB disruption and cognitive impairment by inhibiting TRIC ubiquitination and degradation, likely via interference with VEGF signaling. These results highlight its potential as a therapeutic agent for VCI. Vascular cognitive impairment blood-brain barrier Tubuloside B Tricellulin ubiquitination neuroinflammation chronic cerebral hypoperfusion Full Text Additional Declarations No competing interests reported. Supplementary Files Supplementarymateiral.docx Cite Share Download PDF Status: Published Journal Publication published 29 Dec, 2025 Read the published version in Journal of Neuroinflammation → Version 1 posted Editorial decision: Revision requested 07 Oct, 2025 Reviews received at journal 07 Oct, 2025 Reviews received at journal 30 Sep, 2025 Reviewers agreed at journal 13 Sep, 2025 Reviewers agreed at journal 09 Sep, 2025 Reviewers agreed at journal 08 Sep, 2025 Reviewers agreed at journal 08 Sep, 2025 Reviewers invited by journal 08 Sep, 2025 Editor assigned by journal 08 Sep, 2025 Submission checks completed at journal 07 Sep, 2025 First submitted to journal 07 Sep, 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-7553882","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":514694507,"identity":"be3972be-7b57-46fe-94c9-382f7f061789","order_by":0,"name":"Mingli 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Degradation","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":"[email protected]","identity":"journal-of-neuroinflammation","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"jneu","sideBox":"Learn more about [Journal of Neuroinflammation](http://jneuroinflammation.biomedcentral.com)","snPcode":"12974","submissionUrl":"https://submission.nature.com/new-submission/12974/3","title":"Journal of Neuroinflammation","twitterHandle":"@bmc","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Vascular cognitive impairment, blood-brain barrier, Tubuloside B, Tricellulin, ubiquitination, neuroinflammation, chronic cerebral hypoperfusion","lastPublishedDoi":"10.21203/rs.3.rs-7553882/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7553882/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eBackground and Purpose\u003c/h2\u003e\u003cp\u003eVascular cognitive impairment (VCI) is strongly linked to blood-brain barrier (BBB) dysfunction. This study investigated whether Tubuloside B (Tub-B), a primary bioactive compound of Cistanche tubulosa, ameliorates cognitive deficits by protecting BBB integrity through inhibition of tricellulin (TRIC) ubiquitination and degradation.\u003c/p\u003e\u003ch2\u003eMethods\u003c/h2\u003e\u003cp\u003eThe main plasma metabolites of total glycosides of Cistanche (TCGs) were identified by UPLC-QQQ-MS. A bilateral common carotid artery occlusion (BCCAO) rat model was used to induce chronic cerebral hypoperfusion (CCH). Cognitive function was assessed using the Morris water maze, and BBB permeability was evaluated by Evans blue extravasation. TRIC expression and ubiquitination were analyzed by Western blot, co-immunoprecipitation, and immunohistochemistry. An in vitro oxygen-glucose deprivation (OGD) model in brain microvascular endothelial cells was used to validate the effects on barrier function.\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e\u003cp\u003eTub-B was identified as the major circulating metabolite, accounting for 52.3% \u0026plusmn; 2.1% of plasma components. In CCH rats, Tub-B (20 mg/kg) significantly improved spatial memory, increasing platform crossings (3.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58 vs. CCH, \u003cem\u003eP\u0026thinsp;\u0026lt;\u003c/em\u003e\u0026thinsp;0.05) and reducing Evans blue leakage (0.038\u0026thinsp;\u0026plusmn;\u0026thinsp;0.007 \u0026micro;g/g tissue vs. 0.15\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02 \u0026micro;g/g in CCH, \u003cem\u003eP\u0026thinsp;\u0026lt;\u003c/em\u003e\u0026thinsp;0.001). It also attenuated the downregulation of TRIC and suppressed its ubiquitination. In OGD-injured endothelial cells, Tub-B enhanced barrier integrity and inhibited TRIC ubiquitination.\u003c/p\u003e\u003ch2\u003eConclusion\u003c/h2\u003e\u003cp\u003eTub-B attenuates CCH-induced BBB disruption and cognitive impairment by inhibiting TRIC ubiquitination and degradation, likely via interference with VEGF signaling. These results highlight its potential as a therapeutic agent for VCI.\u003c/p\u003e","manuscriptTitle":"Tubuloside B Ameliorates Blood-Brain Barrier Disruption and Cognitive Deficits after Cerebral Ischemia by Inhibiting TRIC Protein Ubiquitination and Degradation","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-09-15 19:06:18","doi":"10.21203/rs.3.rs-7553882/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-10-07T10:16:07+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-10-07T05:06:43+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-09-30T07:41:05+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"74330380539685741566867522425656825795","date":"2025-09-13T23:32:58+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"302745651327587827447575124678595928495","date":"2025-09-09T08:37:13+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"196140193477034881214768091756647382039","date":"2025-09-08T13:15:59+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"114348087735465929622025446471021954833","date":"2025-09-08T12:20:26+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-09-08T10:45:15+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-09-08T05:59:23+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-09-08T03:45:12+00:00","index":"","fulltext":""},{"type":"submitted","content":"Journal of Neuroinflammation","date":"2025-09-07T04:06:44+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"journal-of-neuroinflammation","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"jneu","sideBox":"Learn more about [Journal of Neuroinflammation](http://jneuroinflammation.biomedcentral.com)","snPcode":"12974","submissionUrl":"https://submission.nature.com/new-submission/12974/3","title":"Journal of Neuroinflammation","twitterHandle":"@bmc","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"b3c74e0a-f038-40a8-8772-379e46e2a1e9","owner":[],"postedDate":"September 15th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2026-01-05T16:07:44+00:00","versionOfRecord":{"articleIdentity":"rs-7553882","link":"https://doi.org/10.1186/s12974-025-03671-5","journal":{"identity":"journal-of-neuroinflammation","isVorOnly":false,"title":"Journal of Neuroinflammation"},"publishedOn":"2025-12-29 15:58:17","publishedOnDateReadable":"December 29th, 2025"},"versionCreatedAt":"2025-09-15 19:06:18","video":"","vorDoi":"10.1186/s12974-025-03671-5","vorDoiUrl":"https://doi.org/10.1186/s12974-025-03671-5","workflowStages":[]},"version":"v1","identity":"rs-7553882","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7553882","identity":"rs-7553882","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}