The Conversion of Evodiamine-induced Hepatotoxicity into a Therapeutic Effect on Colonitis: Insight from the Liver-Gut Axis Mediated by PPAR/NF-κB/ZO-1/caspase-3 Pathway

The Conversion of Evodiamine-induced Hepatotoxicity into a Therapeutic Effect on Colonitis: Insight from the Liver-Gut Axis Mediated by PPAR/NF-κB/ZO-1/caspase-3 Pathway | 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 The Conversion of Evodiamine-induced Hepatotoxicity into a Therapeutic Effect on Colonitis: Insight from the Liver-Gut Axis Mediated by PPAR/NF-κB/ZO-1/caspase-3 Pathway Chongjun Zhao, Qiqi Fan, Ying Dong, Shuang Sun, Yao Zhang, Haiqiang Yao, and 6 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7515324/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 21 Nov, 2025 Read the published version in Chinese Medicine → Version 1 posted 9 You are reading this latest preprint version Abstract Background Evodiamine (EVO) exerts promising therapeutic potential in treating Ulcerative Colitis (UC). However, application in clinical practice is constrained by concerns regarding potential hepatotoxicity. Understanding the mechanisms behind EVO's hepatotoxic effects and its therapeutic benefits is therefore essential to enhance its safe and effective application in clinical practice. Purpose This study aimed to elucidate how the gut-liver axis homeostasis regulates the EVO-induced hepatotoxicity and its therapeutic effects on UC. Methods An integrated experimental strategy utilizing cell, zebrafish, and murine was implemented to assess the hepatotoxic effects of EVO. Transcriptomic and metabolomic analyses in vitro and targeted bile acids (BAs) metabolism studies in vivo were conducted to understand the overall response profile and the underlying mechanisms of hepatotoxicity. Furthermore, the expression patterns of proteins along the gut-liver axis were evaluated under various physiological conditions to identify the relationships contributing to the alleviative effects of UC on EVO-induced hepatotoxicity and the therapeutic effect of EVO on UC. Results High-dose EVO treatment was found to be associated with notable hepatotoxic effects in both in vitro cell models and normal in vivo animals, primarily manifested through disturbances in BAs metabolism, inflammatory responses, and apoptosis. In contrast, in UC models, EVO administration not only effectively improved intestinal structural damage and functional impairments, but also demonstrated minimal hepatotoxicity. Mechanism studies documented that EVO disrupted bile acid metabolism by interfering with BSEP/MRP2/CYP7A1/CYP27A1 pathways, while simultaneously triggering inflammation and apoptosis through PPAR/NF-κB/ZO-1/caspase-3 pathway, ultimately leading to hepatotoxic effects in normal animals. However, UC can mitigate the impact of EVO on protein expression levels in the hepatic, thereby reducing EVO-induced hepatotoxicity. Meanwhile, under UC conditions, EVO can restore the expression levels of relevant proteins in the intestinal tract, thereby maintaining its therapeutic efficacy against UC. Conclusion The hepatotoxicity observed under healthy conditions and the therapeutic effectiveness of EVO against UC are both associated with EVO's regulation of the PPAR/NF-κB/ZO-1/caspase-3 pathway. The influence of EVO on the expression of these key proteins within the gut-liver axis may either counteract or synergistically amplify by different physiological states, potentially leading to varied biological responses across multiple organs. These findings offer valuable insights for the safety assessment and development of traditional Chinese medicines that may pose hepatotoxic risks. Evodiamine Hepatotoxicity Inflammatory Bowel Disease gut-liver axis Full Text Additional Declarations No competing interests reported. Supplementary Files Supplementaryinformation.docx Cite Share Download PDF Status: Published Journal Publication published 21 Nov, 2025 Read the published version in Chinese Medicine → Version 1 posted Editorial decision: Revision requested 28 Sep, 2025 Reviews received at journal 24 Sep, 2025 Reviews received at journal 16 Sep, 2025 Reviewers agreed at journal 14 Sep, 2025 Reviewers agreed at journal 13 Sep, 2025 Reviewers invited by journal 13 Sep, 2025 Editor assigned by journal 11 Sep, 2025 Submission checks completed at journal 11 Sep, 2025 First submitted to journal 02 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-7515324","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":515735839,"identity":"2d58238e-cb6e-4590-9fa9-dd2fe347450d","order_by":0,"name":"Chongjun 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Pathway","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":"chinese-medicine","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"cmed","sideBox":"Learn more about [Chinese Medicine](http://cmjournal.biomedcentral.com)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/cmed/default.aspx","title":"Chinese Medicine","twitterHandle":"@BioMedCentral","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Evodiamine, Hepatotoxicity, Inflammatory Bowel Disease, gut-liver axis","lastPublishedDoi":"10.21203/rs.3.rs-7515324/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7515324/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eBackground\u003c/h2\u003e\u003cp\u003eEvodiamine (EVO) exerts promising therapeutic potential in treating Ulcerative Colitis (UC). However, application in clinical practice is constrained by concerns regarding potential hepatotoxicity. Understanding the mechanisms behind EVO's hepatotoxic effects and its therapeutic benefits is therefore essential to enhance its safe and effective application in clinical practice.\u003c/p\u003e\u003ch2\u003ePurpose\u003c/h2\u003e\u003cp\u003eThis study aimed to elucidate how the gut-liver axis homeostasis regulates the EVO-induced hepatotoxicity and its therapeutic effects on UC.\u003c/p\u003e\u003ch2\u003eMethods\u003c/h2\u003e\u003cp\u003eAn integrated experimental strategy utilizing cell, zebrafish, and murine was implemented to assess the hepatotoxic effects of EVO. Transcriptomic and metabolomic analyses in vitro and targeted bile acids (BAs) metabolism studies in vivo were conducted to understand the overall response profile and the underlying mechanisms of hepatotoxicity. Furthermore, the expression patterns of proteins along the gut-liver axis were evaluated under various physiological conditions to identify the relationships contributing to the alleviative effects of UC on EVO-induced hepatotoxicity and the therapeutic effect of EVO on UC.\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e\u003cp\u003eHigh-dose EVO treatment was found to be associated with notable hepatotoxic effects in both in vitro cell models and normal in vivo animals, primarily manifested through disturbances in BAs metabolism, inflammatory responses, and apoptosis. In contrast, in UC models, EVO administration not only effectively improved intestinal structural damage and functional impairments, but also demonstrated minimal hepatotoxicity. Mechanism studies documented that EVO disrupted bile acid metabolism by interfering with BSEP/MRP2/CYP7A1/CYP27A1 pathways, while simultaneously triggering inflammation and apoptosis through PPAR/NF-κB/ZO-1/caspase-3 pathway, ultimately leading to hepatotoxic effects in normal animals. However, UC can mitigate the impact of EVO on protein expression levels in the hepatic, thereby reducing EVO-induced hepatotoxicity. Meanwhile, under UC conditions, EVO can restore the expression levels of relevant proteins in the intestinal tract, thereby maintaining its therapeutic efficacy against UC.\u003c/p\u003e\u003ch2\u003eConclusion\u003c/h2\u003e\u003cp\u003eThe hepatotoxicity observed under healthy conditions and the therapeutic effectiveness of EVO against UC are both associated with EVO's regulation of the PPAR/NF-κB/ZO-1/caspase-3 pathway. The influence of EVO on the expression of these key proteins within the gut-liver axis may either counteract or synergistically amplify by different physiological states, potentially leading to varied biological responses across multiple organs. These findings offer valuable insights for the safety assessment and development of traditional Chinese medicines that may pose hepatotoxic risks.\u003c/p\u003e","manuscriptTitle":"The Conversion of Evodiamine-induced Hepatotoxicity into a Therapeutic Effect on Colonitis: Insight from the Liver-Gut Axis Mediated by PPAR/NF-κB/ZO-1/caspase-3 Pathway","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-09-19 09:09:29","doi":"10.21203/rs.3.rs-7515324/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-09-28T15:50:31+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-09-24T13:30:08+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-09-16T06:59:05+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"109515540810619406866613080160652683789","date":"2025-09-14T10:27:51+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"273064043447892828744958434458434911004","date":"2025-09-14T01:19:48+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-09-14T01:08:51+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-09-11T16:55:34+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-09-11T16:54:13+00:00","index":"","fulltext":""},{"type":"submitted","content":"Chinese Medicine","date":"2025-09-02T08:22:12+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"chinese-medicine","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"cmed","sideBox":"Learn more about [Chinese Medicine](http://cmjournal.biomedcentral.com)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/cmed/default.aspx","title":"Chinese Medicine","twitterHandle":"@BioMedCentral","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"264537c2-ddd9-4930-8814-5bb98d9419a0","owner":[],"postedDate":"September 19th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2025-11-24T16:11:54+00:00","versionOfRecord":{"articleIdentity":"rs-7515324","link":"https://doi.org/10.1186/s13020-025-01262-3","journal":{"identity":"chinese-medicine","isVorOnly":false,"title":"Chinese Medicine"},"publishedOn":"2025-11-21 15:59:04","publishedOnDateReadable":"November 21st, 2025"},"versionCreatedAt":"2025-09-19 09:09:29","video":"","vorDoi":"10.1186/s13020-025-01262-3","vorDoiUrl":"https://doi.org/10.1186/s13020-025-01262-3","workflowStages":[]},"version":"v1","identity":"rs-7515324","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7515324","identity":"rs-7515324","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}