Multi-omics analysis of chlorogenic acid in alleviating oxidative damage from heat stress in prepubertal boar testes

Multi-omics analysis of chlorogenic acid in alleviating oxidative damage from heat stress in prepubertal boar testes | 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 Multi-omics analysis of chlorogenic acid in alleviating oxidative damage from heat stress in prepubertal boar testes Shaoxuan Zhang, Dali Wang, Jiajia Qi, Jing Li, Simin Liu, Hao Sun, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6918053/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 13 Jan, 2026 Read the published version in Journal of Animal Science and Biotechnology → Version 1 posted 5 You are reading this latest preprint version Abstract Background: Heat stress (HS) can impair boar testicular function, leading to reproductive issues. While chlorogenic acid (CGA) has shown function in mitigating HS-induced damage in various livestock and poultry species. Prepuberty is an important stage of testicular development in boars after birth. However, the protective effect of CGA on testicular HS injury during prepuberty boars and the underlying mechanisms are still not fully understood. Results: In this study, we established an in vivo HS model using prepubertal boars. The results showed that CGA mitigated the HS-induced reduction in testicular tissue antioxidant capacity, decline in Sertoli cell count, and decrease in sperm density post sexual maturation. Utilizing 1H-NMR, RNA-seq and TMT proteomics, we observed alterations in glutathione and other compounds levels in the testicular tissue due to HS in prepubertal boars. Furthermore, the differentially expressed genes influenced by CGA and HS were predominantly associated with the glutathione metabolism pathway. The combined analysis of transcriptomics and proteomics showed that only BLVRA was affected by both HS and CGA when the mRNA and protein levels of a gene showed differential expression with the same trend. Further in vitro studies confirmed that CGA modulated GPX3 expression via BLVRA, impacted GPx activity, and attenuated HS-induced ROS accumulation. Conclusions: Our findings demonstrate that HS in prepubertal boars can impair their ability to produce sperm after sexual maturity. CGA inhibits HS-induced oxidative stress injury in Sertoli cells by restoring GPx activity through BLVRA. This study contributes to elucidating the mechanisms underlying HS effects on prepubertal boar testicular development using multi-omics approaches, thereby laying a foundation for the potential utilization of CGA in swine production. Heat stress Chlorogenic acid Multi omics techniques Testis Oxidative damage Full Text Supplementary Files SupplementTable15.docx SupplementTable6.Correlationcoefficientandpvalue.xlsx SupplementTable7.HeatmapGlutathionemetabolismRNAseq.xlsx SupplementTable8.HeatmapFerroptosisRNAseq.xlsx Supplementfigurelegends.docx Cite Share Download PDF Status: Published Journal Publication published 13 Jan, 2026 Read the published version in Journal of Animal Science and Biotechnology → Version 1 posted Editorial decision: Major revision 13 Oct, 2025 Reviewers agreed at journal 10 Jul, 2025 Reviewers invited by journal 20 Jun, 2025 Editor assigned by journal 18 Jun, 2025 First submitted to journal 17 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-6918053","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":473922498,"identity":"5b62ef1d-31d6-4b90-98af-03eeb3ef9554","order_by":0,"name":"Shaoxuan Zhang","email":"","orcid":"","institution":"Jilin University","correspondingAuthor":false,"prefix":"","firstName":"Shaoxuan","middleName":"","lastName":"Zhang","suffix":""},{"id":473922499,"identity":"ef3872e6-0f09-4325-8518-fbff8a3c825a","order_by":1,"name":"Dali Wang","email":"","orcid":"","institution":"Jilin 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Biotechnology","twitterHandle":"@animalplantsci","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Heat stress, Chlorogenic acid, Multi omics techniques, Testis, Oxidative damage","lastPublishedDoi":"10.21203/rs.3.rs-6918053/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6918053/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"Background: Heat stress (HS) can impair boar testicular function, leading to reproductive issues. While chlorogenic acid (CGA) has shown function in mitigating HS-induced damage in various livestock and poultry species. Prepuberty is an important stage of testicular development in boars after birth. However, the protective effect of CGA on testicular HS injury during prepuberty boars and the underlying mechanisms are still not fully understood.\nResults: In this study, we established an in vivo HS model using prepubertal boars. The results showed that CGA mitigated the HS-induced reduction in testicular tissue antioxidant capacity, decline in Sertoli cell count, and decrease in sperm density post sexual maturation. Utilizing 1H-NMR, RNA-seq and TMT proteomics, we observed alterations in glutathione and other compounds levels in the testicular tissue due to HS in prepubertal boars. Furthermore, the differentially expressed genes influenced by CGA and HS were predominantly associated with the glutathione metabolism pathway. The combined analysis of transcriptomics and proteomics showed that only BLVRA was affected by both HS and CGA when the mRNA and protein levels of a gene showed differential expression with the same trend. Further in vitro studies confirmed that CGA modulated GPX3 expression via BLVRA, impacted GPx activity, and attenuated HS-induced ROS accumulation.\nConclusions: Our findings demonstrate that HS in prepubertal boars can impair their ability to produce sperm after sexual maturity. CGA inhibits HS-induced oxidative stress injury in Sertoli cells by restoring GPx activity through BLVRA. This study contributes to elucidating the mechanisms underlying HS effects on prepubertal boar testicular development using multi-omics approaches, thereby laying a foundation for the potential utilization of CGA in swine production.","manuscriptTitle":"Multi-omics analysis of chlorogenic acid in alleviating oxidative damage from heat stress in prepubertal boar testes","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-06-24 03:53:39","doi":"10.21203/rs.3.rs-6918053/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Major revision","date":"2025-10-13T20:56:26+00:00","index":"","fulltext":""},{"type":"reviewerAgreed","content":"","date":"2025-07-10T10:27:18+00:00","index":0,"fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-06-20T06:29:31+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-06-18T09:58:58+00:00","index":"","fulltext":""},{"type":"submitted","content":"Journal of Animal Science and Biotechnology","date":"2025-06-17T21:42:42+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"journal-of-animal-science-and-biotechnology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"jasb","sideBox":"Learn more about [Journal of Animal Science and Biotechnology](http://jasbsci.biomedcentral.com/)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/jasb/default.aspx","title":"Journal of Animal Science and Biotechnology","twitterHandle":"@animalplantsci","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"eafe66cf-0460-401c-b41f-6940f1f5232e","owner":[],"postedDate":"June 24th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2026-01-19T16:49:28+00:00","versionOfRecord":{"articleIdentity":"rs-6918053","link":"https://doi.org/10.1186/s40104-025-01336-0","journal":{"identity":"journal-of-animal-science-and-biotechnology","isVorOnly":false,"title":"Journal of Animal Science and Biotechnology"},"publishedOn":"2026-01-13 16:30:37","publishedOnDateReadable":"January 13th, 2026"},"versionCreatedAt":"2025-06-24 03:53:39","video":"","vorDoi":"10.1186/s40104-025-01336-0","vorDoiUrl":"https://doi.org/10.1186/s40104-025-01336-0","workflowStages":[]},"version":"v1","identity":"rs-6918053","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6918053","identity":"rs-6918053","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}