ALOX15-Mediated Neuron Ferroptosis Was Involved in Diabetic Peripheral Neuropathic Pain

ALOX15-Mediated Neuron Ferroptosis Was Involved in Diabetic Peripheral Neuropathic Pain | 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 ALOX15-Mediated Neuron Ferroptosis Was Involved in Diabetic Peripheral Neuropathic Pain Zhiye Feng, Zhiqiang Lin, Jian Liu, Xi Chen, Wenxu Yan, Fuye Li, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6058539/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Diabetic peripheral neuropathic pain (DPNP) is a prevalent complication among diabetic patients. Recent studies have confirmed that ferroptosis, an iron-dependent cell death driven by lipid peroxidation, plays a key role in DPNP development. However, biomarkers related to ferroptosis in DPNP remain undiscovered. Core genes related to DPNP were identified using bioinformatics. By assessing the mechanical paw withdrawal threshold (PWT) and thermal paw withdrawal latency (PWL), a mouse model of DPNP was established, along with a high-glucose-stimulated SH-SY5Y cell model. The combination of these two models allows for an in-depth investigation of the molecular biological mechanisms underlying DPNP. The viability and ferroptosis of these cells underwent evaluation via flow cytometry, CCK8 assays, Fe²⁺ assays, along with Western blotting. Differential expression analysis and random forest results identified Arachidonate 15-Lipoxygenase (ALOX15) as a core gene associated with DPNP-related ferroptosis. Subsequently, we verified elevated levels of iron death and ALOX15 expression in neurons using both in vivo and in vitro models. Notably, silencing ALOX15 markedly inhibited high glucose-induced ferroptosis within SH-SY5Y cells and restored neuronal viability. Our findings indicate that ALOX15 promotes the development of DPNP by mediating neuronal ferroptosis, providing new insights into the role of ALOX15 as a biomarker for DPNP while proposing innovative strategies to prevent and treat DPNP. Diabetic peripheral neuropathic pain ALOX15 Ferroptosis Machine Learning Full Text Additional Declarations No competing interests reported. Supplementary Files WesternBlot.pdf Cite Share Download PDF Status: Posted Version 1 posted 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-6058539","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":417931326,"identity":"9f9d2e0a-aaa3-475b-a779-5a483951d468","order_by":0,"name":"Zhiye Feng","email":"","orcid":"","institution":"Southern Medical University","correspondingAuthor":false,"prefix":"","firstName":"Zhiye","middleName":"","lastName":"Feng","suffix":""},{"id":417931327,"identity":"694cf32c-fde4-43f4-a862-1e44809a007a","order_by":1,"name":"Zhiqiang Lin","email":"","orcid":"","institution":"Southern Medical 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Pain","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"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":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Diabetic peripheral neuropathic pain, ALOX15, Ferroptosis, Machine Learning","lastPublishedDoi":"10.21203/rs.3.rs-6058539/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6058539/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"Diabetic peripheral neuropathic pain (DPNP) is a prevalent complication among diabetic patients. Recent studies have confirmed that ferroptosis, an iron-dependent cell death driven by lipid peroxidation, plays a key role in DPNP development. However, biomarkers related to ferroptosis in DPNP remain undiscovered. Core genes related to DPNP were identified using bioinformatics. By assessing the mechanical paw withdrawal threshold (PWT) and thermal paw withdrawal latency (PWL), a mouse model of DPNP was established, along with a high-glucose-stimulated SH-SY5Y cell model. The combination of these two models allows for an in-depth investigation of the molecular biological mechanisms underlying DPNP. The viability and ferroptosis of these cells underwent evaluation via flow cytometry, CCK8 assays, Fe²⁺ assays, along with Western blotting. Differential expression analysis and random forest results identified Arachidonate 15-Lipoxygenase (ALOX15) as a core gene associated with DPNP-related ferroptosis. Subsequently, we verified elevated levels of iron death and ALOX15 expression in neurons using both in vivo and in vitro models. Notably, silencing ALOX15 markedly inhibited high glucose-induced ferroptosis within SH-SY5Y cells and restored neuronal viability. Our findings indicate that ALOX15 promotes the development of DPNP by mediating neuronal ferroptosis, providing new insights into the role of ALOX15 as a biomarker for DPNP while proposing innovative strategies to prevent and treat DPNP.","manuscriptTitle":"ALOX15-Mediated Neuron Ferroptosis Was Involved in Diabetic Peripheral Neuropathic Pain","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-02-21 11:53:34","doi":"10.21203/rs.3.rs-6058539/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"81bff18d-0bc3-4b36-abf1-2ffb5294dbf5","owner":[],"postedDate":"February 21st, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-04-28T05:08:50+00:00","versionOfRecord":[],"versionCreatedAt":"2025-02-21 11:53:34","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6058539","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6058539","identity":"rs-6058539","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}