L-Carnitine protects against methotrexate-induced hepatotoxicity via regulation of apoptosis and metabolic pathways

L-Carnitine protects against methotrexate-induced hepatotoxicity via regulation of apoptosis and metabolic pathways | 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 L-Carnitine protects against methotrexate-induced hepatotoxicity via regulation of apoptosis and metabolic pathways Sakir PEKGOZ, Zafer USTA, Yagmur KARA, Ozlem OZMEN This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7429081/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 24 Mar, 2026 Read the published version in Naunyn-Schmiedeberg's Archives of Pharmacology → Version 1 posted 7 You are reading this latest preprint version Abstract Background Methotrexate (MTX) is a widely used chemotherapeutic and immunosuppressive drug; however, its clinical application is limited by dose-dependent hepatotoxicity mediated by oxidative stress, mitochondrial dysfunction, and apoptosis. Objective This study aimed to evaluate the hepatoprotective potential of L-carnitine (LCR), a mitochondrial cofactor with antioxidant and anti-apoptotic properties, in a rat model of MTX-induced hepatotoxicity. Methods Twenty-four male Wistar rats were randomly divided into four groups: Control, MTX (20 mg/kg, i.p., single dose), LCR (100 mg/kg/day, oral), and MTX + LCR. After 10 days, liver tissues were examined histopathologically, immunohistochemically for CD38 and GLUT1, and by qPCR for BAX and BCL2 expression. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were also measured. Results MTX administration elevated ALT and AST levels, caused histological injury, upregulated CD38 and GLUT1 expression, and dysregulated apoptosis-related genes (BAX↑, BCL2↓). LCR co-treatment improved hepatic histoarchitecture, reduced CD38 and GLUT1 expression, partially normalized BAX/BCL2 levels, and tended to restore serum enzyme activity. Conclusion L-carnitine protects against MTX-induced hepatotoxicity by reducing oxidative and metabolic stress and modulating apoptosis. These findings support the potential of L-carnitine as an adjuvant in MTX-based chemotherapy to enhance hepatic safety and minimize off-target toxicity. Methotrexate L-carnitine hepatotoxicity oxidative stress apoptosis CD38 GLUT1 BAX/BCL2 Full Text Additional Declarations No competing interests reported. Cite Share Download PDF Status: Published Journal Publication published 24 Mar, 2026 Read the published version in Naunyn-Schmiedeberg's Archives of Pharmacology → Version 1 posted Editorial decision: Revision requested 23 Sep, 2025 Reviews received at journal 20 Sep, 2025 Reviewers agreed at journal 15 Sep, 2025 Reviewers invited by journal 09 Sep, 2025 Editor assigned by journal 21 Aug, 2025 Submission checks completed at journal 21 Aug, 2025 First submitted to journal 21 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. We do this by developing innovative software and high quality services for the global research community. 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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-7429081","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":515234098,"identity":"d7fc5b04-abf1-45ee-9724-fd21e7c6fc97","order_by":0,"name":"Sakir PEKGOZ","email":"","orcid":"","institution":"Isparta Health Services Vocational School, Süleyman Demirel University","correspondingAuthor":false,"prefix":"","firstName":"Sakir","middleName":"","lastName":"PEKGOZ","suffix":""},{"id":515234099,"identity":"b984f8a9-b845-4541-bf8d-44df5e4fb7b0","order_by":1,"name":"Zafer USTA","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA/klEQVRIiWNgGAWjYNCCCgYGAzDjgAQ/A4yNBzA2MJwBKwMyDkhINoDZCQS0MLbBtTAQ1iI/I/35g5/ztsmZix0+/uDHGQsJBvbmbRKMP+7htmJGjmFj77bbxpaz0xIbe25ISDDwHCuTYEgoxqmFWSKHsYF32+3EDbdzDBt4PkjUMUjkmAG14HYZm0T6w8a/c27Xb7id/7HxzwegLfJv8GvhkUgwbOZtuJ1gcDuHsZkH5DAJHvxaJHjeGM6WOXbbcMPtNCDjjIQEG09asUVCGm4t8u3pDz6+qbktb3A7Gcg4VifBz354440PNri1YPEdiCBFwygYBaNgFIwCTAAArStWKLdQe9AAAAAASUVORK5CYII=","orcid":"","institution":"Mehmet Akif Ersoy University","correspondingAuthor":true,"prefix":"","firstName":"Zafer","middleName":"","lastName":"USTA","suffix":""},{"id":515234100,"identity":"885de2ec-94c5-4da4-be11-28564229fc60","order_by":2,"name":"Yagmur KARA","email":"","orcid":"","institution":"Isparta City Hospital","correspondingAuthor":false,"prefix":"","firstName":"Yagmur","middleName":"","lastName":"KARA","suffix":""},{"id":515234101,"identity":"b75b86cf-d99d-469e-9731-038615f6f8b9","order_by":3,"name":"Ozlem OZMEN","email":"","orcid":"","institution":"Mehmet Akif Ersoy University","correspondingAuthor":false,"prefix":"","firstName":"Ozlem","middleName":"","lastName":"OZMEN","suffix":""}],"badges":[],"createdAt":"2025-08-21 20:23:12","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-7429081/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7429081/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s00210-026-05196-x","type":"published","date":"2026-03-24T16:12:45+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":105754996,"identity":"5ef647ab-350d-40ce-8b38-762eaf66f95d","added_by":"auto","created_at":"2026-03-30 16:23:59","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":998142,"visible":true,"origin":"","legend":"","description":"","filename":"Manuscript21082025MTXLCR.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7429081/v1_covered_b6bb03a2-f9bf-4190-af58-4258c2b97c22.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"L-Carnitine protects against methotrexate-induced hepatotoxicity via regulation of apoptosis and metabolic pathways","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":"naunyn-schmiedebergs-archives-of-pharmacology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"nsap","sideBox":"Learn more about [Naunyn-Schmiedeberg's Archives of Pharmacology](https://www.springer.com/journal/210)","snPcode":"210","submissionUrl":"https://submission.nature.com/new-submission/210/3","title":"Naunyn-Schmiedeberg's Archives of Pharmacology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Methotrexate, L-carnitine, hepatotoxicity, oxidative stress, apoptosis, CD38, GLUT1, BAX/BCL2","lastPublishedDoi":"10.21203/rs.3.rs-7429081/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7429081/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eBackground\u003c/h2\u003e\u003cp\u003eMethotrexate (MTX) is a widely used chemotherapeutic and immunosuppressive drug; however, its clinical application is limited by dose-dependent hepatotoxicity mediated by oxidative stress, mitochondrial dysfunction, and apoptosis.\u003c/p\u003e\u003ch2\u003eObjective\u003c/h2\u003e\u003cp\u003eThis study aimed to evaluate the hepatoprotective potential of L-carnitine (LCR), a mitochondrial cofactor with antioxidant and anti-apoptotic properties, in a rat model of MTX-induced hepatotoxicity.\u003c/p\u003e\u003ch2\u003eMethods\u003c/h2\u003e\u003cp\u003eTwenty-four male Wistar rats were randomly divided into four groups: Control, MTX (20 mg/kg, i.p., single dose), LCR (100 mg/kg/day, oral), and MTX\u0026thinsp;+\u0026thinsp;LCR. After 10 days, liver tissues were examined histopathologically, immunohistochemically for CD38 and GLUT1, and by qPCR for BAX and BCL2 expression. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were also measured.\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e\u003cp\u003eMTX administration elevated ALT and AST levels, caused histological injury, upregulated CD38 and GLUT1 expression, and dysregulated apoptosis-related genes (BAX\u0026uarr;, BCL2\u0026darr;). LCR co-treatment improved hepatic histoarchitecture, reduced CD38 and GLUT1 expression, partially normalized BAX/BCL2 levels, and tended to restore serum enzyme activity.\u003c/p\u003e\u003ch2\u003eConclusion\u003c/h2\u003e\u003cp\u003eL-carnitine protects against MTX-induced hepatotoxicity by reducing oxidative and metabolic stress and modulating apoptosis. These findings support the potential of L-carnitine as an adjuvant in MTX-based chemotherapy to enhance hepatic safety and minimize off-target toxicity.\u003c/p\u003e","manuscriptTitle":"L-Carnitine protects against methotrexate-induced hepatotoxicity via regulation of apoptosis and metabolic pathways","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-09-16 17:41:55","doi":"10.21203/rs.3.rs-7429081/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-09-23T09:56:33+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-09-20T16:54:23+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"217511030195266813447911386268398587320","date":"2025-09-15T10:16:26+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-09-09T07:42:47+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-08-22T01:54:49+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-08-22T01:53:48+00:00","index":"","fulltext":""},{"type":"submitted","content":"Naunyn-Schmiedeberg's Archives of Pharmacology","date":"2025-08-21T20:13:56+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"naunyn-schmiedebergs-archives-of-pharmacology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"nsap","sideBox":"Learn more about [Naunyn-Schmiedeberg's Archives of Pharmacology](https://www.springer.com/journal/210)","snPcode":"210","submissionUrl":"https://submission.nature.com/new-submission/210/3","title":"Naunyn-Schmiedeberg's Archives of Pharmacology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"c01464b0-8c09-4e5b-862d-f0283df88ba1","owner":[],"postedDate":"September 16th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2026-03-30T16:18:58+00:00","versionOfRecord":{"articleIdentity":"rs-7429081","link":"https://doi.org/10.1007/s00210-026-05196-x","journal":{"identity":"naunyn-schmiedebergs-archives-of-pharmacology","isVorOnly":false,"title":"Naunyn-Schmiedeberg's Archives of Pharmacology"},"publishedOn":"2026-03-24 16:12:45","publishedOnDateReadable":"March 24th, 2026"},"versionCreatedAt":"2025-09-16 17:41:55","video":"","vorDoi":"10.1007/s00210-026-05196-x","vorDoiUrl":"https://doi.org/10.1007/s00210-026-05196-x","workflowStages":[]},"version":"v1","identity":"rs-7429081","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7429081","identity":"rs-7429081","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}