Protective effect of high-intensity aerobic training and Crocin on testicular oxidative stress and apoptosis in healthy male Wistar rats treated with doxorubicin

Protective effect of high-intensity aerobic training and Crocin on testicular oxidative stress and apoptosis in healthy male Wistar rats treated with doxorubicin | 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 Article Protective effect of high-intensity aerobic training and Crocin on testicular oxidative stress and apoptosis in healthy male Wistar rats treated with doxorubicin Maryam Zilaei Bouri, Mohsen Davoodi, Ali Delpisheh, Shahla Dehghan Ghahfarokhi, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8097930/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 16 You are reading this latest preprint version Abstract Chemotherapy-induced testicular toxicity remains a significant limitation in male reproductive health, with oxidative stress and apoptosis as key mediators. This study investigated the effects of HIAT and Crocin on doxorubicin (DX)- induced testicular damage. Thirty-five male Wistar rats were allocated into seven groups (n=5): healthy control (HC), doxorubicin control (DXC), DX + Crocin 10 mg/kg (DX-C10), DX + Crocin 50 mg/kg (DX-C50), DX + HIAT, DX + HIAT + C10, and DX + HIAT + C50. Doxorubicin was administered at 2 mg/kg/week for seven weeks. HIAT consisted of 2–8 × 2-min intervals at 80–110% vV̇ O₂max, 5 days/week. Oxidative stress and apoptosis were quantified via MDA levels, and Caspase-3/9 expression (ELISA, qPCR), and testicular morphology was assessed using H&E and DAPI staining. Doxorubicin increased oxidative stress and apoptosis, by elevated MDA and Caspase-3/9 (P < 0.01), and reduced seminiferous tubule diameter (P < 0.01). HIAT, Crocin, and their combination significantly mitigated these effects (P < 0.01). The DX + HIAT + C50 group demonstrated the greatest attenuation of apoptotic markers and histopathological damage, whereas Crocin 50 mg/kg alone reduced MDA levels (P<0.05). These findings suggest that Crocin plus HIAT may serve as a potential strategy to mitigate chemotherapy-related reproductive damage. Biological sciences/Biochemistry Biological sciences/Cell biology Biological sciences/Drug discovery Biological sciences/Physiology Caspases Chemical Compound Chemotherapy Exercise reproductive health Full Text Additional Declarations No competing interests reported. Cite Share Download PDF Status: Under Review Version 1 posted Editorial decision: Revision requested 30 Dec, 2025 Reviews received at journal 30 Dec, 2025 Reviews received at journal 28 Dec, 2025 Reviewers agreed at journal 28 Dec, 2025 Reviews received at journal 27 Dec, 2025 Reviewers agreed at journal 26 Dec, 2025 Reviewers agreed at journal 25 Dec, 2025 Reviewers agreed at journal 24 Dec, 2025 Reviewers agreed at journal 23 Dec, 2025 Reviewers agreed at journal 23 Dec, 2025 Reviewers agreed at journal 23 Dec, 2025 Reviewers invited by journal 23 Dec, 2025 Editor invited by journal 15 Dec, 2025 Editor assigned by journal 15 Dec, 2025 Submission checks completed at journal 11 Dec, 2025 First submitted to journal 11 Dec, 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-8097930","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":564699442,"identity":"1ee1a536-b6e9-4eec-b3ba-c3c7374ba0b5","order_by":0,"name":"Maryam Zilaei Bouri","email":"","orcid":"","institution":"Tarbiat Modares University","correspondingAuthor":false,"prefix":"","firstName":"Maryam","middleName":"Zilaei","lastName":"Bouri","suffix":""},{"id":564699443,"identity":"ab2d2427-6d1f-4079-9347-9fc219787453","order_by":1,"name":"Mohsen Davoodi","email":"","orcid":"","institution":"Iz.C, Islamic Azad 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doxorubicin","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"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":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Caspases, Chemical 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This study investigated the effects of HIAT and Crocin on doxorubicin (DX)- induced testicular damage. Thirty-five male Wistar rats were allocated into seven groups (n=5): healthy control (HC), doxorubicin control (DXC), DX + Crocin 10 mg/kg (DX-C10), DX + Crocin 50 mg/kg (DX-C50), DX + HIAT, DX + HIAT + C10, and DX + HIAT + C50. Doxorubicin was administered at 2 mg/kg/week for seven weeks. HIAT consisted of 2–8 × 2-min intervals at 80–110% vV̇ O₂max, 5 days/week. Oxidative stress and apoptosis were quantified via MDA levels, and Caspase-3/9 expression (ELISA, qPCR), and testicular morphology was assessed using H\u0026amp;E and DAPI staining. Doxorubicin increased oxidative stress and apoptosis, by elevated MDA and Caspase-3/9 (P \u0026lt; 0.01), and reduced seminiferous tubule diameter (P \u0026lt; 0.01). HIAT, Crocin, and their combination significantly mitigated these effects (P \u0026lt; 0.01). The DX + HIAT + C50 group demonstrated the greatest attenuation of apoptotic markers and histopathological damage, whereas Crocin 50 mg/kg alone reduced MDA levels (P\u0026lt;0.05). 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