Hydrogen-rich water improves skeletal muscle damage in mice with Duchenne muscular dystrophy

Hydrogen-rich water improves skeletal muscle damage in mice with Duchenne muscular dystrophy | 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 Hydrogen-rich water improves skeletal muscle damage in mice with Duchenne muscular dystrophy Jie Shao, Binyan Wan, Tian Gao, Yongguang Shi, Yingfeng Zhu, Xu’en Yu This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7380679/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 Objective To study the beneficial effect of hydrogen-rich water (HRW) on skeletal muscle damage in mice with Duchenne muscular dystrophy (DMD). Methods Wild-type C57BL/6J mice served as the normal control group, and mdx mice were randomly divided into the model and HRW groups, with six mice in each group. The HRW treatment lasted for 8 weeks. Grip strength and rod turning tests were employed to assess the motor ability. Creatine kinase (CK) levels were measured using an Enzyme linked immunosorbent assay (ELISA). Histological staining of skeletal muscle was conducted to observe the pathological changes of skeletal muscle tissue, and immunohistochemistry and immunofluorescence staining were used to detect the expression of dystrophin protein. Transmission electron microscopy was used to observe the ultrastructure of the mitochondria in each group. Results Compared with the model group, the grip value and drop time of mice in the HRW group increased by 59.01% and 54.32% ( P < 0.001), respectively, and the CK level decreased by 26.25% ( P < 0.01). Histological staining revealed certain improvements in the architecture and enzymology of skeletal muscle in the HRW group, with decreased muscle interstitium and increased muscle content. Ultrastructural observation demonstrated that the mitochondrial morphology was improved and that vacuolation was reduced, and immunological detection indicated slight expression of dystrophin protein. Conclusion The results showed that HRW improved the muscle damage in DMD model mice. Health sciences/Anatomy Biological sciences/Cell biology Health sciences/Diseases Health sciences/Medical research Biological sciences/Physiology Duchenne muscular dystrophy (DMD) mdx mice hydrogen-rich water (HRW) skeletal muscle mitochondria Full Text Additional Declarations No competing interests reported. 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. 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-7380679","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":537609146,"identity":"1144b342-af69-4e4e-97e8-85e46b2ce499","order_by":0,"name":"Jie Shao","email":"","orcid":"","institution":"Institute of Neurology of Anhui University of Chinese Medicine","correspondingAuthor":false,"prefix":"","firstName":"Jie","middleName":"","lastName":"Shao","suffix":""},{"id":537609147,"identity":"cf39ae70-dacc-4dfa-b125-cf3b685b2082","order_by":1,"name":"Binyan Wan","email":"","orcid":"","institution":"Institute of Neurology of Anhui University of 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dystrophy","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":"Duchenne muscular dystrophy (DMD), mdx mice, hydrogen-rich water (HRW), skeletal muscle, mitochondria","lastPublishedDoi":"10.21203/rs.3.rs-7380679/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7380679/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eObjective\u003c/h2\u003e\u003cp\u003eTo study the beneficial effect of hydrogen-rich water (HRW) on skeletal muscle damage in mice with Duchenne muscular dystrophy (DMD).\u003c/p\u003e\u003ch2\u003eMethods\u003c/h2\u003e\u003cp\u003eWild-type C57BL/6J mice served as the normal control group, and mdx mice were randomly divided into the model and HRW groups, with six mice in each group. The HRW treatment lasted for 8 weeks. Grip strength and rod turning tests were employed to assess the motor ability. Creatine kinase (CK) levels were measured using an Enzyme linked immunosorbent assay (ELISA). Histological staining of skeletal muscle was conducted to observe the pathological changes of skeletal muscle tissue, and immunohistochemistry and immunofluorescence staining were used to detect the expression of dystrophin protein. Transmission electron microscopy was used to observe the ultrastructure of the mitochondria in each group.\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e\u003cp\u003eCompared with the model group, the grip value and drop time of mice in the HRW group increased by 59.01% and 54.32% (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001), respectively, and the CK level decreased by 26.25% (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.01). Histological staining revealed certain improvements in the architecture and enzymology of skeletal muscle in the HRW group, with decreased muscle interstitium and increased muscle content. Ultrastructural observation demonstrated that the mitochondrial morphology was improved and that vacuolation was reduced, and immunological detection indicated slight expression of dystrophin protein.\u003c/p\u003e\u003ch2\u003eConclusion\u003c/h2\u003e\u003cp\u003eThe results showed that HRW improved the muscle damage in DMD model mice.\u003c/p\u003e","manuscriptTitle":"Hydrogen-rich water improves skeletal muscle damage in mice with Duchenne muscular dystrophy","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-10-31 16:34:10","doi":"10.21203/rs.3.rs-7380679/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":"6ecd9c22-89a0-4015-a3eb-015bd5d0436a","owner":[],"postedDate":"October 31st, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":57175359,"name":"Health sciences/Anatomy"},{"id":57175360,"name":"Biological sciences/Cell biology"},{"id":57175361,"name":"Health sciences/Diseases"},{"id":57175362,"name":"Health sciences/Medical research"},{"id":57175363,"name":"Biological sciences/Physiology"}],"tags":[],"updatedAt":"2025-11-10T17:38:38+00:00","versionOfRecord":[],"versionCreatedAt":"2025-10-31 16:34:10","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7380679","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7380679","identity":"rs-7380679","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}