Cholesterol orchestrates skin fibrosis via the CH25H-CYP7B1-SMAD2/3 axis

Cholesterol orchestrates skin fibrosis via the CH25H-CYP7B1-SMAD2/3 axis | 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 Cholesterol orchestrates skin fibrosis via the CH25H-CYP7B1-SMAD2/3 axis Jiefeng Huang, Shuangmeng Jia, Zixin Cai, Yingjia Zhu, Wuyan Lu, and 9 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8366356/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 8 You are reading this latest preprint version Abstract Skin fibrosis is a pathological condition characterized by excessive deposition of extracellular matrix (ECM) components, primarily collagen, which leads to compromised tissue function. Although skin fibrosis is currently considered to be associated with metabolic disorders, the role of cholesterol metabolism in the pathogenesis of skin fibrosis has not been fully elucidated. scRNA-seq and RNA-seq analyses revealed that key modulators involved in cholesterol metabolism, such as lectin-type oxidized low-density lipoprotein receptor 1 (LOX1), cholesterol 25-hydroxylase (CH25H), and 25-hydroxycholesterol 7α-hydroxylase (CYP7B1), were upregulated in human keloid samples. Mendelian randomization (MR) analysis demonstrated a causal relationship between cholesterol levels and dermal fibrosis. We further confirmed that cholesterol levels were elevated in fibrotic skin and were positively correlated with the severity of fibrosis. Moreover, the subcutaneous injection of cholesterol significantly increased the expression of fibrotic indicators, including Col1a1 , Col3a1 , Fn1 , and α-SMA , and promoted both spontaneous and bleomycin (BLM)-induced skin fibrosis. Notably, this cholesterol-induced increase in the expression of phospho-Smad2/3 and fibrotic indicators was significantly attenuated when Ch25h was knocked down. Furthermore, the administration of tauroursodeoxycholic acid (TUDCA), which promotes cholesterol efflux, resulted in a remarkable reduction in intracellular cholesterol levels and ameliorated skin fibrosis in BLM-induced mice. Our results demonstrate that the CH25H-CYP7B1-SMAD2/3 axis is crucial in the pathogenesis of skin fibrosis. TUDCA treatment reduced cholesterol accumulation and mitigated the progression of skin fibrosis, providing a promising therapeutic strategy. Cholesterol Skin fibrosis CH25H-CYP7B1 axis SMAD2/3 Systemic sclerosis Full Text Additional Declarations No competing interests reported. Supplementary Files SupplementaryMaterial.docx Westernblot.pdf Cite Share Download PDF Status: Under Review Version 1 posted Editorial decision: Revision requested 11 Mar, 2026 Reviews received at journal 09 Mar, 2026 Reviews received at journal 13 Feb, 2026 Reviewers agreed at journal 13 Feb, 2026 Reviewers agreed at journal 05 Feb, 2026 Reviewers invited by journal 02 Feb, 2026 Submission checks completed at journal 27 Jan, 2026 First submitted to journal 21 Jan, 2026 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. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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-8366356","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":585154244,"identity":"53ceafb4-87df-43da-a8c5-5d97d8d67bbf","order_by":0,"name":"Jiefeng Huang","email":"","orcid":"","institution":"Shanghai Tenth People's Hospital","correspondingAuthor":false,"prefix":"","firstName":"Jiefeng","middleName":"","lastName":"Huang","suffix":""},{"id":585154245,"identity":"24f1dcec-5e72-44d9-b167-42555545461b","order_by":1,"name":"Shuangmeng Jia","email":"","orcid":"","institution":"Tongji University","correspondingAuthor":false,"prefix":"","firstName":"Shuangmeng","middleName":"","lastName":"Jia","suffix":""},{"id":585154246,"identity":"ce5c2f33-ce06-439a-b041-8d63c78d374e","order_by":2,"name":"Zixin Cai","email":"","orcid":"","institution":"Shanghai Tenth People's Hospital","correspondingAuthor":false,"prefix":"","firstName":"Zixin","middleName":"","lastName":"Cai","suffix":""},{"id":585154247,"identity":"a1e9fec3-4def-4e50-9a1d-b36093b01a24","order_by":3,"name":"Yingjia Zhu","email":"","orcid":"","institution":"Tongji University","correspondingAuthor":false,"prefix":"","firstName":"Yingjia","middleName":"","lastName":"Zhu","suffix":""},{"id":585154248,"identity":"4ae9030b-cdf6-44f9-8be9-7b5aa82598be","order_by":4,"name":"Wuyan Lu","email":"","orcid":"","institution":"Shanghai Tenth People's Hospital","correspondingAuthor":false,"prefix":"","firstName":"Wuyan","middleName":"","lastName":"Lu","suffix":""},{"id":585154249,"identity":"3098b680-3b97-4da3-8d16-ff73714928bc","order_by":5,"name":"Ke Fang","email":"","orcid":"","institution":"Shanghai Tenth People's Hospital","correspondingAuthor":false,"prefix":"","firstName":"Ke","middleName":"","lastName":"Fang","suffix":""},{"id":585154250,"identity":"0bfa1bf8-62db-4b0e-a631-ae8a89a63477","order_by":6,"name":"Yishu Lu","email":"","orcid":"","institution":"Tongji University","correspondingAuthor":false,"prefix":"","firstName":"Yishu","middleName":"","lastName":"Lu","suffix":""},{"id":585154251,"identity":"d8299204-baca-4f6d-b4df-33ee4c6c7b9c","order_by":7,"name":"Linxiao Li","email":"","orcid":"","institution":"Tongji University","correspondingAuthor":false,"prefix":"","firstName":"Linxiao","middleName":"","lastName":"Li","suffix":""},{"id":585154252,"identity":"7c42a01f-5aab-4cc9-ab7c-5c1fd51e91f5","order_by":8,"name":"Yitong Ji","email":"","orcid":"","institution":"Tongji University","correspondingAuthor":false,"prefix":"","firstName":"Yitong","middleName":"","lastName":"Ji","suffix":""},{"id":585154254,"identity":"2261a6c9-aa24-446a-b59d-f8a19953f38b","order_by":9,"name":"Fengting Niu","email":"","orcid":"","institution":"Shanghai Tenth People's Hospital","correspondingAuthor":false,"prefix":"","firstName":"Fengting","middleName":"","lastName":"Niu","suffix":""},{"id":585154256,"identity":"373bf7e0-0a11-4408-b0b7-6ed1c2a6a468","order_by":10,"name":"Guangpeng Liu","email":"","orcid":"","institution":"Shanghai Tenth People's Hospital","correspondingAuthor":false,"prefix":"","firstName":"Guangpeng","middleName":"","lastName":"Liu","suffix":""},{"id":585154258,"identity":"7a413741-d3ea-4e93-9164-ca5e18f99cb7","order_by":11,"name":"Tian Li","email":"","orcid":"","institution":"Tianjin Key Laboratory of Acute Abdomen Disease-Associated Organ Injury and ITCWM Repair","correspondingAuthor":false,"prefix":"","firstName":"Tian","middleName":"","lastName":"Li","suffix":""},{"id":585154259,"identity":"795d8d95-b489-4a7d-b396-66e4e74c04b3","order_by":12,"name":"Lei Cui","email":"","orcid":"","institution":"Stem Cell Translational Research Center","correspondingAuthor":false,"prefix":"","firstName":"Lei","middleName":"","lastName":"Cui","suffix":""},{"id":585154260,"identity":"2f20680e-3a7b-40f7-9104-a7f480d57c90","order_by":13,"name":"Shuaijun Li","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA/klEQVRIiWNgGAWjYBACPmYGNiAlwczAwHwAJMAM5uLTwobQwpbYQJwWMAIDHsMGmCh+Lezszx7z1FiwG9zu+f7g445adj4G5oO3eRjs8nA7jMfcmOeYBLPBnbMbG2eeOQ50GFuyNQ9DcjEeLWzSPGxALTdyNzbzth0DauExk+ZhOJDYgFML+zNpnn8gLTkPoVr4vxHQwmAmzdsG1sII1FIDsoWNgBYeM8m5fRLMkjfSDGfObDvAzMbMZmw5xyAZpxZ+/uPPJN58q0vmu5H84MPHtrpk+fbmhzfeVNjh1AIDyVD6cDIDM4g2IKAeCOygdJ0dXmWjYBSMglEwIgEAdClGEdachykAAAAASUVORK5CYII=","orcid":"","institution":"Shanghai Tenth People's Hospital","correspondingAuthor":true,"prefix":"","firstName":"Shuaijun","middleName":"","lastName":"Li","suffix":""}],"badges":[],"createdAt":"2025-12-15 12:53:09","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-8366356/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-8366356/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":101943184,"identity":"8d7c2d78-7c20-4c7f-a1ac-8a376db0e496","added_by":"auto","created_at":"2026-02-05 09:41:01","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2760924,"visible":true,"origin":"","legend":"","description":"","filename":"Manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8366356/v1_covered_4a4188d1-fea0-4182-aebe-f8b1358d2ae6.pdf"},{"id":101904782,"identity":"d3bdca21-b1c7-4e3c-af7d-9d6ba089502a","added_by":"auto","created_at":"2026-02-04 20:27:19","extension":"docx","order_by":0,"title":"","display":"","copyAsset":false,"role":"supplement","size":8262934,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementaryMaterial.docx","url":"https://assets-eu.researchsquare.com/files/rs-8366356/v1/91b0e2c25f4fafee6c3d24b5.docx"},{"id":101904781,"identity":"79ad0171-1298-470b-a310-25a5c1ac8214","added_by":"auto","created_at":"2026-02-04 20:27:19","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":1274618,"visible":true,"origin":"","legend":"","description":"","filename":"Westernblot.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8366356/v1/b428858694b61c07723c1385.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Cholesterol orchestrates skin fibrosis via the CH25H-CYP7B1-SMAD2/3 axis","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":false,"email":"[email protected]","identity":"cellular-and-molecular-biology-letters","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"cmbl","sideBox":"Learn more about [Cellular \u0026 Molecular Biology Letters](http://cmbl.biomedcentral.com)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/CMBL/default.aspx","title":"Cellular \u0026 Molecular Biology Letters","twitterHandle":"@OACellBiology","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Cholesterol, Skin fibrosis, CH25H-CYP7B1 axis, SMAD2/3, Systemic sclerosis","lastPublishedDoi":"10.21203/rs.3.rs-8366356/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8366356/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eSkin fibrosis is a pathological condition characterized by excessive deposition of extracellular matrix (ECM) components, primarily collagen, which leads to compromised tissue function. Although skin fibrosis is currently considered to be associated with metabolic disorders, the role of cholesterol metabolism in the pathogenesis of skin fibrosis has not been fully elucidated. scRNA-seq and RNA-seq analyses revealed that key modulators involved in cholesterol metabolism, such as lectin-type oxidized low-density lipoprotein receptor 1 (LOX1), cholesterol 25-hydroxylase (CH25H), and 25-hydroxycholesterol 7α-hydroxylase (CYP7B1), were upregulated in human keloid samples. Mendelian randomization (MR) analysis demonstrated a causal relationship between cholesterol levels and dermal fibrosis. We further confirmed that cholesterol levels were elevated in fibrotic skin and were positively correlated with the severity of fibrosis. Moreover, the subcutaneous injection of cholesterol significantly increased the expression of fibrotic indicators, including \u003cem\u003eCol1a1\u003c/em\u003e, \u003cem\u003eCol3a1\u003c/em\u003e, \u003cem\u003eFn1\u003c/em\u003e, and \u003cem\u003eα-SMA\u003c/em\u003e, and promoted both spontaneous and bleomycin (BLM)-induced skin fibrosis. Notably, this cholesterol-induced increase in the expression of phospho-Smad2/3 and fibrotic indicators was significantly attenuated when \u003cem\u003eCh25h\u003c/em\u003e was knocked down. Furthermore, the administration of tauroursodeoxycholic acid (TUDCA), which promotes cholesterol efflux, resulted in a remarkable reduction in intracellular cholesterol levels and ameliorated skin fibrosis in BLM-induced mice. Our results demonstrate that the CH25H-CYP7B1-SMAD2/3 axis is crucial in the pathogenesis of skin fibrosis. TUDCA treatment reduced cholesterol accumulation and mitigated the progression of skin fibrosis, providing a promising therapeutic strategy.\u003c/p\u003e","manuscriptTitle":"Cholesterol orchestrates skin fibrosis via the CH25H-CYP7B1-SMAD2/3 axis","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-02-04 20:27:14","doi":"10.21203/rs.3.rs-8366356/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2026-03-11T09:11:58+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-03-09T16:00:45+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-02-13T14:22:48+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"267469011181953741659693682349133315327","date":"2026-02-13T14:03:48+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"235237751519290733227628531438858605860","date":"2026-02-05T13:56:47+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2026-02-02T14:58:46+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2026-01-27T05:01:28+00:00","index":"","fulltext":""},{"type":"submitted","content":"Cellular \u0026 Molecular Biology Letters","date":"2026-01-21T15:19:00+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":false,"email":"[email protected]","identity":"cellular-and-molecular-biology-letters","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"cmbl","sideBox":"Learn more about [Cellular \u0026 Molecular Biology Letters](http://cmbl.biomedcentral.com)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/CMBL/default.aspx","title":"Cellular \u0026 Molecular Biology Letters","twitterHandle":"@OACellBiology","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"a2d1a40c-e8a3-43aa-a26a-814f770ba064","owner":[],"postedDate":"February 4th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2026-03-27T09:38:41+00:00","versionOfRecord":[],"versionCreatedAt":"2026-02-04 20:27:14","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8366356","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8366356","identity":"rs-8366356","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}