Quercetin ameliorates cyclophosphamide-induced premature ovarian insufficiency by modulating SIRT1/HIF-1α pathway

Quercetin ameliorates cyclophosphamide-induced premature ovarian insufficiency by modulating SIRT1/HIF-1α 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 Quercetin ameliorates cyclophosphamide-induced premature ovarian insufficiency by modulating SIRT1/HIF-1α pathway Mei Wang, Hongling Chen, Shaozi Lin, Zuyu Meng, Yiyao Zhang, Fang Liu, and 7 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6972937/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 13 Mar, 2026 Read the published version in Journal of Ovarian Research → Version 1 posted 4 You are reading this latest preprint version Abstract Background Premature ovarian insufficiency (POI) is characterized by complex etiologies and currently lacks effective treatments, posing significant threats to women’s physical and psychological health. Quercetin (QUE), a bioactive flavonoid inherent in traditional herbs such as Flos Sophorae and Cuscuta chinensis, demonstrates potent redox-modulating and anti-senescence capacities. This investigation validates its therapeutic efficacy against POI, specifically elucidating granulosa cell-centric mechanisms. Methods POI mouse models and human ovarian granulosa cells KGN injury models were induced through the administration of cyclophosphamide (CTX). Subsequently, these models were treated with drugs such as QUE. Morphological changes in the ovaries of the mice were observed, and vaginal smears were conducted to monitor the estrus cycles and patterns of the mice. ELISA was employed to detect sex hormone levels in various mouse groups, while Hematoxylin-eosin staining Staining(H&E) staining was utilized to assess the status of ovarian follicles at different stages. Moving forward, immunohistochemical analysis and Western blot were conducted to detect the expression levels of Sirtuin 1 (SIRT1), Hypoxia-Inducible Factor 1 Alpha (HIF-1α), and other apoptosis-related proteins. Ultimately, methods such as Terminal deoxynucleotidyl transferase dUTP Nick-End Labeling (TUNEL) assay, β -galactosidase staining, flow cytometry for apoptosis detection, and 5-Ethynyl-2′-deoxyuridine (EdU) Assay assay were utilized to evaluate the apoptosis, senescence, and proliferation of mouse ovarian tissues and human ovarian granulosa cells KGN. Results QUE ameliorated CTX-induced ovarian atrophy and follicular atresia by upregulating SIRT1 expression. Furthermore, QUE partially restored estrous cyclicity, normalized sex hormone levels, and improved follicular counts through SIRT1/HIF-1α modulation. Concurrently, QUE suppressed apoptosis in both murine and human granulosa cells via the SIRT1/HIF-1α pathway. Conclusion Our findings demonstrate that QUE enhances ovarian reserve by inhibiting granulosa cells apoptosis via SIRT1/HIF-1α signaling, highlighting its therapeutic potential for POI management. Premature ovarian insufficiency quercetin SIRT1 HIF-1α Full Text Additional Declarations No competing interests reported. Supplementary Files Fig.S1.pptx Supplementary2.docx Cite Share Download PDF Status: Published Journal Publication published 13 Mar, 2026 Read the published version in Journal of Ovarian Research → Version 1 posted Reviewers invited by journal 13 Jul, 2025 Editor assigned by journal 02 Jul, 2025 Submission checks completed at journal 01 Jul, 2025 First submitted to journal 25 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. 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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-6972937","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":484832066,"identity":"726a385d-c6aa-4a52-878f-04f59ff3b10e","order_by":0,"name":"Mei Wang","email":"","orcid":"","institution":"Jinan University","correspondingAuthor":false,"prefix":"","firstName":"Mei","middleName":"","lastName":"Wang","suffix":""},{"id":484832067,"identity":"b7aa27d4-3b81-4591-9262-37d9c9d7dcf8","order_by":1,"name":"Hongling Chen","email":"","orcid":"","institution":"Jinan 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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":"journal-of-ovarian-research","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"jovr","sideBox":"Learn more about [Journal of Ovarian Research](http://ovarianresearch.biomedcentral.com)","snPcode":"13048","submissionUrl":"https://submission.nature.com/new-submission/13048/3","title":"Journal of Ovarian Research","twitterHandle":"@BioMedCentral","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Premature ovarian insufficiency, quercetin, SIRT1, HIF-1α","lastPublishedDoi":"10.21203/rs.3.rs-6972937/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6972937/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eBackground\u003c/h2\u003e\u003cp\u003ePremature ovarian insufficiency (POI) is characterized by complex etiologies and currently lacks effective treatments, posing significant threats to women\u0026rsquo;s physical and psychological health. Quercetin (QUE), a bioactive flavonoid inherent in traditional herbs such as Flos Sophorae and Cuscuta chinensis, demonstrates potent redox-modulating and anti-senescence capacities. This investigation validates its therapeutic efficacy against POI, specifically elucidating granulosa cell-centric mechanisms.\u003c/p\u003e\u003ch2\u003eMethods\u003c/h2\u003e\u003cp\u003ePOI mouse models and human ovarian granulosa cells KGN injury models were induced through the administration of cyclophosphamide (CTX). Subsequently, these models were treated with drugs such as QUE. Morphological changes in the ovaries of the mice were observed, and vaginal smears were conducted to monitor the estrus cycles and patterns of the mice. ELISA was employed to detect sex hormone levels in various mouse groups, while Hematoxylin-eosin staining Staining(H\u0026amp;E) staining was utilized to assess the status of ovarian follicles at different stages. Moving forward, immunohistochemical analysis and Western blot were conducted to detect the expression levels of Sirtuin 1 (SIRT1), Hypoxia-Inducible Factor 1 Alpha (HIF-1α), and other apoptosis-related proteins. Ultimately, methods such as Terminal deoxynucleotidyl transferase dUTP Nick-End Labeling (TUNEL) assay, \u003cem\u003eβ\u003c/em\u003e-galactosidase staining, flow cytometry for apoptosis detection, and 5-Ethynyl-2\u0026prime;-deoxyuridine (EdU) Assay assay were utilized to evaluate the apoptosis, senescence, and proliferation of mouse ovarian tissues and human ovarian granulosa cells KGN.\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e\u003cp\u003eQUE ameliorated CTX-induced ovarian atrophy and follicular atresia by upregulating SIRT1 expression. Furthermore, QUE partially restored estrous cyclicity, normalized sex hormone levels, and improved follicular counts through SIRT1/HIF-1α modulation. Concurrently, QUE suppressed apoptosis in both murine and human granulosa cells via the SIRT1/HIF-1α pathway.\u003c/p\u003e\u003ch2\u003eConclusion\u003c/h2\u003e\u003cp\u003eOur findings demonstrate that QUE enhances ovarian reserve by inhibiting granulosa cells apoptosis via SIRT1/HIF-1α signaling, highlighting its therapeutic potential for POI management.\u003c/p\u003e","manuscriptTitle":"Quercetin ameliorates cyclophosphamide-induced premature ovarian insufficiency by modulating SIRT1/HIF-1α pathway","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-07-15 05:06:10","doi":"10.21203/rs.3.rs-6972937/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"reviewersInvited","content":"","date":"2025-07-13T09:20:35+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-07-03T00:48:49+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-07-01T12:14:31+00:00","index":"","fulltext":""},{"type":"submitted","content":"Journal of Ovarian Research","date":"2025-06-25T09:20:45+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"journal-of-ovarian-research","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"jovr","sideBox":"Learn more about [Journal of Ovarian Research](http://ovarianresearch.biomedcentral.com)","snPcode":"13048","submissionUrl":"https://submission.nature.com/new-submission/13048/3","title":"Journal of Ovarian Research","twitterHandle":"@BioMedCentral","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"fa063b45-dcae-4444-9fcd-1aecb0ceafc6","owner":[],"postedDate":"July 15th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2026-03-16T16:14:36+00:00","versionOfRecord":{"articleIdentity":"rs-6972937","link":"https://doi.org/10.1186/s13048-026-02052-7","journal":{"identity":"journal-of-ovarian-research","isVorOnly":false,"title":"Journal of Ovarian Research"},"publishedOn":"2026-03-13 15:59:29","publishedOnDateReadable":"March 13th, 2026"},"versionCreatedAt":"2025-07-15 05:06:10","video":"","vorDoi":"10.1186/s13048-026-02052-7","vorDoiUrl":"https://doi.org/10.1186/s13048-026-02052-7","workflowStages":[]},"version":"v1","identity":"rs-6972937","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6972937","identity":"rs-6972937","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}