Hydrolyzed Peptides from Pichia pastoris Improve Sperm Quality in Male Rats by Modulating the PI3K/AKT Signalling Pathway

Hydrolyzed Peptides from Pichia pastoris Improve Sperm Quality in Male Rats by Modulating the PI3K/AKT Signalling 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 Article Hydrolyzed Peptides from Pichia pastoris Improve Sperm Quality in Male Rats by Modulating the PI3K/AKT Signalling Pathway Junyu Xue, Juyue Luo, Xutong Tian, Yaping Zhao, Chia Hau Lee, and 4 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7823492/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 Oxidative stress is one of the critical factors contributing to the rising prevalence of male infertility. Hydrolyzed peptides from Pichia pastoris (HPP), a major component of Pichia pastoris hydrolysate, have an underestimated impact on the male reproductive system. This study aimed to systematically evaluate the potential of HPP to improve sperm quality. Results revealed that HPP enhanced sperm quality and serum testosterone levels in rats, with no adverse impact on body weight or the genital weight index. HPP increased the diameter of seminiferous tubules, yet no significant changes were observed in the expression levels of blood-testis-barrier (BTB)-related genes (zonula occludens-1, Occludin, Connexin-43, and N-cadherin). Transcriptomics, non-target metabolomics, Western blotting, and Quantitative Real-time PCR analyses revealed that HPP-treated rats exhibited significantly upregulated antioxidant enzymes (nuclear factor erythroid-2-related factor 2 (Nrf2) and Heme Oxygenase-1 (HO-1)) expression, decreased reactive oxygen species (ROS) contents, and inhibited the PI3K/AKT signaling pathway. Moreover, we found that taurine and hypotaurine metabolism might be potential metabolic targets for improving sperm quality. Taken together, these results highlight HPP as a promising nutraceutical supplement for improving sperm quality. The antioxidant mechanism of HPP via the regulation of the PI3K/AKT signaling pathway lays a foundation for further research into nutrient-based interventions for male infertility. Biological sciences/Biochemistry Biological sciences/Cell biology Health sciences/Diseases Biological sciences/Molecular biology Biological sciences/Physiology antioxidant hydrolysed peptides sperm quality male infertility nutrient Full Text Additional Declarations No competing interests reported. Supplementary Files originalWB.xlsx 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-7823492","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":540759520,"identity":"f8955d39-7213-4b80-a130-8483ab46d7df","order_by":0,"name":"Junyu Xue","email":"","orcid":"","institution":"The First Affiliated Hospital, College of Clinical Medicine, Henan University of Science and Technology","correspondingAuthor":false,"prefix":"","firstName":"Junyu","middleName":"","lastName":"Xue","suffix":""},{"id":540759521,"identity":"87a8ed79-6d93-4a67-9c9c-c9a09b0a4a35","order_by":1,"name":"Juyue Luo","email":"","orcid":"","institution":"College 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Pathway\u003c/p\u003e","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":true,"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":"antioxidant, hydrolysed peptides, sperm quality, male infertility, nutrient","lastPublishedDoi":"10.21203/rs.3.rs-7823492/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7823492/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eOxidative stress is one of the critical factors contributing to the rising prevalence of male infertility. Hydrolyzed peptides from \u003cem\u003ePichia pastoris\u003c/em\u003e (HPP), a major component of \u003cem\u003ePichia pastoris\u003c/em\u003e hydrolysate, have an underestimated impact on the male reproductive system. This study aimed to systematically evaluate the potential of HPP to improve sperm quality. Results revealed that HPP enhanced sperm quality and serum testosterone levels in rats, with no adverse impact on body weight or the genital weight index. HPP increased the diameter of seminiferous tubules, yet no significant changes were observed in the expression levels of blood-testis-barrier (BTB)-related genes (zonula occludens-1, Occludin, Connexin-43, and N-cadherin). Transcriptomics, non-target metabolomics, Western blotting, and Quantitative Real-time PCR analyses revealed that HPP-treated rats exhibited significantly upregulated antioxidant enzymes (nuclear factor erythroid-2-related factor 2 (Nrf2) and Heme Oxygenase-1 (HO-1)) expression, decreased reactive oxygen species (ROS) contents, and inhibited the PI3K/AKT signaling pathway. Moreover, we found that taurine and hypotaurine metabolism might be potential metabolic targets for improving sperm quality. Taken together, these results highlight HPP as a promising nutraceutical supplement for improving sperm quality. The antioxidant mechanism of HPP via the regulation of the PI3K/AKT signaling pathway lays a foundation for further research into nutrient-based interventions for male infertility.\u003c/p\u003e","manuscriptTitle":"Hydrolyzed Peptides from Pichia pastoris Improve Sperm Quality in Male Rats by Modulating the PI3K/AKT Signalling Pathway","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-11-07 10:34:04","doi":"10.21203/rs.3.rs-7823492/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":"fdb0d5df-aa7d-495f-b4fe-e150f7a3d348","owner":[],"postedDate":"November 7th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":57606331,"name":"Biological sciences/Biochemistry"},{"id":57606332,"name":"Biological sciences/Cell biology"},{"id":57606333,"name":"Health sciences/Diseases"},{"id":57606334,"name":"Biological sciences/Molecular biology"},{"id":57606335,"name":"Biological sciences/Physiology"}],"tags":[],"updatedAt":"2025-12-11T11:38:41+00:00","versionOfRecord":[],"versionCreatedAt":"2025-11-07 10:34:04","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7823492","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7823492","identity":"rs-7823492","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}