Caffeoylquinic acids and their microbiota-derived metabolites ameliorate hepatic lipid metabolism disorders and egg yolk lipid composition via the gut-liver axis

Caffeoylquinic acids and their microbiota-derived metabolites ameliorate hepatic lipid metabolism disorders and egg yolk lipid composition via the gut-liver 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 Caffeoylquinic acids and their microbiota-derived metabolites ameliorate hepatic lipid metabolism disorders and egg yolk lipid composition via the gut-liver axis Wenjie Tian, Qianhai Zhang, Sichao Wang, Gerard Bryan Gonzales, and 7 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7836529/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 Background: Fatty Liver Hemorrhagic Syndrome (FLHS) is currently the most prevalent lipid metabolism disorders disease in laying hens. Although FLHS has been studied for a long time, there are currently no effective therapeutic strategies. The regulation of gut microbiota is an important strategy and novel target for the treatment of fatty liver disease with good prospects. Caffeoylquinic acids (CQAs) may be potential gut microbiota modulators due to their extremely low bioavailability. However, the effects of CQAs on gut microbiota structure, hepatic lipid metabolism, and egg yolk fatty acids composition in laying hens, as well as their underlying mechanisms, remain poorly understood. Results: In this study, we used aged laying hens as a model to investigate the effect of CQAs on ameliorating FLHS, and the treatment groups were added with differential proportions of Artemisia argyi extract (SAE, rich in CQAs), and chlorogenic acid (CGA, used to verify the effects of CQAs). Our multi-omics approach (metagenomics, transcriptomics, and metabolite analysis) yielded several key findings: (1) CQAs significantly ameliorated FLHS, improving laying performance and egg quality while reducing hepatic lipid storage; (2) Metagenomic analysis identified Bacteroides and Phocaeicola as key bacterial genera mediating the anti-FLHS effects; (3) Microbial metabolites analysis revealed that CQAs supplementation reshaped the gut microbial ecosystem, increasing the levels of short-chain fatty acids (SCFAs) and specific microbial catabolites of CQAs (e.g., caffeic acid (CA), 3-(3'-hydroxyphenyl)propionic acid (3-HPP), 3-hydroxybenzoic acid (3-HBA), syringic acid, and vanillic acid); (4) Hepatic transcriptomics revealed that CQAs downregulated key lipogenic genes (e.g., ACLY , FASN , ACACA , SCD1 ) and upregulated genes involved in lipid transport and oxidation (e.g., ApoB , CPT1A , FABPs , CD36 , ACOX1 , SCP2 ); (5) Mechanistically, CQAs treatment markedly activated the ADPN-AMPK-PPARα pathway, thereby ameliorating lipid metabolism disorders and improving laying performance. Conclusions: Our findings indicate that CQAs and their microbiota-derived metabolites ameliorate hepatic lipid metabolism disorders and egg yolk lipid composition via the gut-liver axis. Crucially, we traced these metabolites from the gut to the systemic circulation and liver, establishing them as the primary effectors that activate the hepatic ADPN-AMPK-PPARα pathway. This activation coordinately downregulates lipogenesis and upregulates fatty acid transport and oxidation, thereby alleviating hepatic steatosis. This coordinated response alleviates hepatic steatosis and, notably, redirects lipids towards the ovary, enhancing the deposition of nutritionally beneficial fatty acids in egg yolk, which is a finding with significant implications for egg quality. Our work highlights gut microbiota metabolites as a promising therapeutic target for lipid metabolism disorders and transforms the understanding of how low-bioavailability substances exert their systemic effects. Caffeoylquinic acids FLHS Lipid metabolism Gut-liver axis Full Text Additional Declarations No competing interests reported. Supplementary Files Supplementaryfigure11.10.docx SupplementaryTable2025.11.10.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-7836529","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":563492241,"identity":"3ee13c77-5638-4cd9-971b-8e6a04a063a3","order_by":0,"name":"Wenjie Tian","email":"","orcid":"","institution":"State Key Laboratory of Animal Nutrition and Feeding, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences","correspondingAuthor":false,"prefix":"","firstName":"Wenjie","middleName":"","lastName":"Tian","suffix":""},{"id":563492242,"identity":"961d71dd-6f5c-40af-adcf-5cfec5e8bdbe","order_by":1,"name":"Qianhai Zhang","email":"","orcid":"","institution":"State Key Laboratory of Animal Nutrition and Feeding, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences","correspondingAuthor":false,"prefix":"","firstName":"Qianhai","middleName":"","lastName":"Zhang","suffix":""},{"id":563492243,"identity":"fbe3b0f1-e92b-4915-9df3-2b97bcba9d58","order_by":2,"name":"Sichao Wang","email":"","orcid":"","institution":"State Key Laboratory of Animal Nutrition and Feeding, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences","correspondingAuthor":false,"prefix":"","firstName":"Sichao","middleName":"","lastName":"Wang","suffix":""},{"id":563492244,"identity":"6e9e42f4-5176-46e7-a2f6-afbd8785add3","order_by":3,"name":"Gerard Bryan Gonzales","email":"","orcid":"","institution":"Ghent 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axis","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":"Caffeoylquinic acids, FLHS, Lipid metabolism, Gut-liver axis","lastPublishedDoi":"10.21203/rs.3.rs-7836529/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7836529/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground:\u003c/strong\u003e Fatty Liver Hemorrhagic Syndrome (FLHS) is currently the most prevalent lipid metabolism disorders disease in laying hens. Although FLHS has been studied for a long time, there are currently no effective therapeutic strategies. The regulation of gut microbiota is an important strategy and novel target for the treatment of fatty liver disease with good prospects. Caffeoylquinic acids (CQAs) may be potential gut microbiota modulators due to their extremely low bioavailability. However, the effects of CQAs on gut microbiota structure, hepatic lipid metabolism, and egg yolk fatty acids composition in laying hens, as well as their underlying mechanisms, remain poorly understood.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults:\u003c/strong\u003e In this study, we used aged laying hens as a model to investigate the effect of CQAs on ameliorating FLHS, and the treatment groups were added with differential proportions of \u003cem\u003eArtemisia argyi\u003c/em\u003eextract (SAE, rich in CQAs), and chlorogenic acid (CGA, used to verify the effects of CQAs). Our multi-omics approach (metagenomics, transcriptomics, and metabolite analysis) yielded several key findings: (1) CQAs significantly ameliorated FLHS, improving laying performance and egg quality while reducing hepatic lipid storage; (2) Metagenomic analysis identified \u003cem\u003eBacteroides\u003c/em\u003eand \u003cem\u003ePhocaeicola\u003c/em\u003e as key bacterial genera mediating the anti-FLHS effects; (3) Microbial metabolites analysis revealed that CQAs supplementation reshaped the gut microbial ecosystem, increasing the levels of short-chain fatty acids (SCFAs) and specific microbial catabolites of CQAs (e.g., caffeic acid (CA), 3-(3'-hydroxyphenyl)propionic acid (3-HPP), 3-hydroxybenzoic acid (3-HBA), syringic acid, and vanillic acid); (4) Hepatic transcriptomics revealed that CQAs downregulated key lipogenic genes (e.g., \u003cem\u003eACLY\u003c/em\u003e, \u003cem\u003eFASN\u003c/em\u003e, \u003cem\u003eACACA\u003c/em\u003e, \u003cem\u003eSCD1\u003c/em\u003e) and upregulated genes involved in lipid transport and oxidation (e.g., \u003cem\u003eApoB\u003c/em\u003e, \u003cem\u003eCPT1A\u003c/em\u003e, \u003cem\u003eFABPs\u003c/em\u003e, \u003cem\u003eCD36\u003c/em\u003e,\u003cem\u003e ACOX1\u003c/em\u003e,\u003cem\u003e SCP2\u003c/em\u003e); (5) Mechanistically, CQAs treatment markedly activated the ADPN-AMPK-PPARα pathway, thereby ameliorating lipid metabolism disorders and improving laying performance.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusions:\u003c/strong\u003e Our findings indicate that CQAs and their microbiota-derived metabolites ameliorate hepatic lipid metabolism disorders and egg yolk lipid composition via the gut-liver axis. Crucially, we traced these metabolites from the gut to the systemic circulation and liver, establishing them as the primary effectors that activate the hepatic ADPN-AMPK-PPARα pathway. This activation coordinately downregulates lipogenesis and upregulates fatty acid transport and oxidation, thereby alleviating hepatic steatosis. This coordinated response alleviates hepatic steatosis and, notably, redirects lipids towards the ovary, enhancing the deposition of nutritionally beneficial fatty acids in egg yolk, which is a finding with significant implications for egg quality. Our work highlights gut microbiota metabolites as a promising therapeutic target for lipid metabolism disorders and transforms the understanding of how low-bioavailability substances exert their systemic effects.\u003c/p\u003e","manuscriptTitle":"Caffeoylquinic acids and their microbiota-derived metabolites ameliorate hepatic lipid metabolism disorders and egg yolk lipid composition via the gut-liver axis","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-12-22 15:17:01","doi":"10.21203/rs.3.rs-7836529/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":"2c6585b5-8c6f-4e46-8f32-dbef78a35783","owner":[],"postedDate":"December 22nd, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2026-02-08T09:39:37+00:00","versionOfRecord":[],"versionCreatedAt":"2025-12-22 15:17:01","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7836529","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7836529","identity":"rs-7836529","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}