6-Gingerol inhibition of inflammation and apoptosis through the activation of SIRT1 in Balb/c mice

Targeting early events in hepatocarcinogenesis: anti-inflammatory and SIRT1-mediated effects of 6-Gingerol in a murine model | 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 Targeting early events in hepatocarcinogenesis: anti-inflammatory and SIRT1-mediated effects of 6-Gingerol in a murine model Ikenna C. Maduako, Ebenezer O. Farombi This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6706491/v2 This work is licensed under a CC BY 4.0 License Status: Posted Version 2 posted You are reading this latest preprint version Show more versions Abstract Background: Persistent inflammation, compensatory cell proliferation, and gene regulation through epigenetic modifications are frequently linked to hepatocellular carcinoma (HCC) development. Therefore, natural compounds that can suppress inflammatory processes are vital in improving the pathogenesis of diseases triggered by chronic inflammation. Although Sirtuin-1—a nuclear sirtuin known for its anti-inflammatory roles in diverse tissues and cells including macrophages—has gained interest, its specific contribution in HCC remains insufficiently explored. Methods: This study employed a mouse model to induce liver cancer using diethylnitrosamine (DEN) and 2-acetylaminofluorene (2AAF). A total of forty male BALB/c mice, weighing around 18 ± 2 g, were divided to four groups of ten. Group 1 was orally treated with a dose of corn oil, while Group 2 received a single intraperitoneal dose of DEN at 75 mg/kg, followed by 2AAF at 200 mg/kg dietary plan. Meanwhile, Group 3 was solely treated with 6-gingerol at 100 mg/kg. Lastly, Group 4 received 6-gingerol for 7 days prior to DEN + 2AAF exposure, with treatment maintained for 20 weeks. Results: Administration of 6-gingerol conferred protective effects in HCC-induced mice by lowering the levels of TNF-α, IL-1β, COX-2, and iNOS. Its anti-inflammatory potential was further demonstrated through reduced expression of NF-κB and BCL2 in liver tissues, alongside increased levels of p53 and SIRT-1. Conclusion: The findings suggest that 6-gingerol counteracts DEN+2AAF-induced liver cancer by downregulating inflammatory mediators and promoting apoptosis through SIRT-1 activation. Cancer Biology Biochemical Research Methods Pathology 6-Gingerol SIRT1 HCC inflammation apoptosis mice Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Full Text Additional Declarations The authors declare no competing interests. Supplementary Files HIGHLIGHTS.doc Highlight GRAPHICALABSTRACT.ppt Graphical Abstract Cite Share Download PDF Status: Posted Version 2 posted You are reading this latest preprint version Show more versions 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-6706491","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":464208757,"identity":"f33be580-891a-4a9a-b45a-e64a3b86797e","order_by":0,"name":"Ikenna C. 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Farombi","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABC0lEQVRIiWNgGAWjYDACdjDJDEQHQIwDPPwMDGz4tTAzMDaAtbAlQLRINpCqhcHgAAEt/MzMzx/z7rCWY2BjfvbhY84dGeMbyc8efKhgkOcXO4BVi2Qzm2Ez75l0YwY2NuOZM7c94zG7kWZuOOMMg+HM2QlYtRgcZgBqaTuc2CDfYMzMu+0wUEuCmTRvG0OCwW1cWtg/grTUN7CxfwZrMZ6R/o2AFh6wLQkMbDwQWwwkcvDbItnMUzhzblu6YQMbTzEjyC8SZ96USc44I4HTL/zs7Rs+vG2zlmdgY9/M8HHbHXv+9vRtEh8qbOT5pbFrgQP7AzCWAFilBH7laBYfIKhkFIyCUTAKRhYAANc5V7HeowjSAAAAAElFTkSuQmCC","orcid":"","institution":"","correspondingAuthor":true,"prefix":"","firstName":"Ebenezer","middleName":"O.","lastName":"Farombi","suffix":""}],"badges":[],"createdAt":"2025-05-20 10:05:02","currentVersionCode":2,"declarations":{"humanSubjects":false,"vertebrateSubjects":true,"conflictsOfInterestStatement":false,"humanSubjectEthicalGuidelines":false,"humanSubjectConsent":false,"humanSubjectClinicalTrial":false,"humanSubjectCaseReport":false,"vertebrateSubjectEthicalGuidelines":true},"doi":"10.21203/rs.3.rs-6706491/v2","doiUrl":"https://doi.org/10.21203/rs.3.rs-6706491/v2","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":84010197,"identity":"74dcb080-4d5d-43f5-a2b0-dec574c77a89","added_by":"auto","created_at":"2025-06-05 16:20:43","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":42144,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of 6-Gingerol on the activities of CAT, SOD, GST and GSH level in liver of DEN+2AAF-treated mice respectively for 20 weeks. Each bar represents mean ± SD of 10 mice. a: Values differ significantly from control (p \u0026lt; 0.05). b: Values differ significantly from DEN+2AAF group (p \u0026lt; 0.05).\u003c/p\u003e","description":"","filename":"Slide1.png","url":"https://assets-eu.researchsquare.com/files/rs-6706491/v2/bd33075f566665be8f0c3576.png"},{"id":84010198,"identity":"5488d722-fc00-41bb-b2c6-ed7e5b3592ee","added_by":"auto","created_at":"2025-06-05 16:20:43","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":36588,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of 6-Gingerol on H2O2, LPO, GPX and XO activities in liver of DEN+2AAF-treated mice for 20 weeks. Each bar represents mean ± SD of 10 mice. a: Values differ significantly from control (p \u0026lt; 0.05). b: Values differ significantly from DEN+2AAF group (p \u0026lt; 0.05).\u003c/p\u003e","description":"","filename":"Slide2.png","url":"https://assets-eu.researchsquare.com/files/rs-6706491/v2/527dad6b8bf70bb0e69402df.png"},{"id":84010199,"identity":"d54694a5-10ed-4e8d-800f-5141142f7008","added_by":"auto","created_at":"2025-06-05 16:20:43","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":126927,"visible":true,"origin":"","legend":"\u003cp\u003ea: Effect of 6-Gingerol on NO level and MPO activity in liver of DEN+2AAF-treated mice for 20 weeks. Each bar represents mean ± SD of 10 mice. a: Values differ significantly from control (p \u0026lt; 0.05). b: Values differ significantly from DEN+2AAF group (p \u0026lt; 0.05).\u003c/p\u003e\n\u003cp\u003eb: Effects of 6G on pro-inflammatory enzymes iNOS and COX-2 and cytokines TNF-α and IL-1β levels in the liver of mice exposed to DEN+2AAF for 20 weeks. Each bar represents mean ± SD of 10 mice. a: Values differ significantly from control (p \u0026lt; 0.05).b: Values differ significantly from DEN+2AAF group (p \u0026lt; 0.05).\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-6706491/v2/87a63912ac177816e243ad8f.png"},{"id":84011062,"identity":"7e4e40b5-8100-4432-b725-79f80e5be055","added_by":"auto","created_at":"2025-06-05 16:28:43","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":566584,"visible":true,"origin":"","legend":"\u003cp\u003eImmunofluorescence stain of mice liver section for the effect of 6G on NF-κB and Sirt-1 protein expression following DEN+2AAF exposure for 20 weeks. a: Values differ significantly from control (p \u0026lt; 0.05).b: Values differ significantly from DEN+2AAF group (p \u0026lt; 0.05). (x40).\u003c/p\u003e","description":"","filename":"Slide5.png","url":"https://assets-eu.researchsquare.com/files/rs-6706491/v2/2b5d664e49b49bb70494b12c.png"},{"id":84011064,"identity":"997d72de-8fa5-4945-97b4-76b937288b5b","added_by":"auto","created_at":"2025-06-05 16:28:43","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":602194,"visible":true,"origin":"","legend":"\u003cp\u003eRepresentative photomicrographs of histopathological examination of experimental mice liver after 20 weeks treatment. H\u0026amp;E (X400). (A) Control orally administered olive oil (2mL/kg): showed normal liver architecture. (B) DEN+2-AAF: showed fibrosis and inflammatory cells, mainly lymphocytes and macrophages in the distorted portal space. (C) 6[G]: showed normal liver architecture. (D) 6[G]+DEN+2-AAF: showed restored integrity of the liver with mild hepatic congestion.\u003c/p\u003e","description":"","filename":"Slide6.png","url":"https://assets-eu.researchsquare.com/files/rs-6706491/v2/4cebad1ae912412c3411522b.png"},{"id":84011431,"identity":"c0c2a5f3-6206-4374-98ad-cadd04209410","added_by":"auto","created_at":"2025-06-05 16:36:43","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":745572,"visible":true,"origin":"","legend":"\u003cp\u003eImmunohistochemistry stain of mice liver section for the effect of 6G on Ki-67 and cyclin D1 protein expression following DEN+2AAF exposure for 20 weeks. a: Values differ significantly from control (p \u0026lt; 0.05).b: Values differ significantly from DEN+2AAF group (p \u0026lt; 0.05). (x40). Brown colour indicates positive staining for Ki-67 and CCND1 antibody on 5 μm section of paraffin fixed mice liver.\u003c/p\u003e","description":"","filename":"Slide7.png","url":"https://assets-eu.researchsquare.com/files/rs-6706491/v2/7ef79f6f4db93f5fa7bf4bc7.png"},{"id":84011066,"identity":"a6f054a4-68a3-43ae-8a26-6a4428be0fc8","added_by":"auto","created_at":"2025-06-05 16:28:43","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":758825,"visible":true,"origin":"","legend":"\u003cp\u003eImmunohistochemistry and immunofluorescence stain of mice liver section for the effect of 6G on BCL2 and P-53 protein expression following DEN+2AAF exposure for 20 weeks. a: Values differ significantly from control (p \u0026lt; 0.05).b: Values differ significantly from DEN+2AAF group (p \u0026lt; 0.05). (x40). Brown colour indicates positive staining for BCL2 antibody on 5 μm section of paraffin fixed mice liver.\u003c/p\u003e","description":"","filename":"Slide8.png","url":"https://assets-eu.researchsquare.com/files/rs-6706491/v2/61b59ca94705fb58c26041ee.png"},{"id":84010201,"identity":"6a57c748-8076-437b-9905-e104a9debecf","added_by":"auto","created_at":"2025-06-05 16:20:43","extension":"doc","order_by":4,"title":"","display":"","copyAsset":false,"role":"supplement","size":31232,"visible":true,"origin":"","legend":"\u003cp\u003eHighlight\u003c/p\u003e","description":"","filename":"HIGHLIGHTS.doc","url":"https://assets-eu.researchsquare.com/files/rs-6706491/v2/64309eea472641a1a4001b88.doc"},{"id":84010210,"identity":"632906ba-1a32-4fde-98e6-911047b88ff9","added_by":"auto","created_at":"2025-06-05 16:20:43","extension":"ppt","order_by":5,"title":"","display":"","copyAsset":false,"role":"supplement","size":2345472,"visible":true,"origin":"","legend":"\u003cp\u003eGraphical Abstract\u003c/p\u003e","description":"","filename":"GRAPHICALABSTRACT.ppt","url":"https://assets-eu.researchsquare.com/files/rs-6706491/v2/03974f7e82900c96846c2624.ppt"}],"financialInterests":"The authors declare no competing interests.","formattedTitle":"\u003cp\u003e\u003cstrong\u003eTargeting early events in hepatocarcinogenesis: anti-inflammatory and SIRT1-mediated effects of 6-Gingerol in a murine model\u003c/strong\u003e\u003c/p\u003e","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"University of Ibadan","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"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":"6-Gingerol, SIRT1, HCC, inflammation, apoptosis, mice","lastPublishedDoi":"10.21203/rs.3.rs-6706491/v2","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6706491/v2","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eBackground: Persistent inflammation, compensatory cell proliferation, and gene regulation through epigenetic modifications are frequently linked to hepatocellular carcinoma (HCC) development. Therefore, natural compounds that can suppress inflammatory processes are vital in improving the pathogenesis of diseases triggered by chronic inflammation. Although Sirtuin-1—a nuclear sirtuin known for its anti-inflammatory roles in diverse tissues and cells including macrophages—has gained interest, its specific contribution in HCC remains insufficiently explored.\u003c/p\u003e\n\u003cp\u003eMethods: This study employed a mouse model to induce liver cancer using diethylnitrosamine (DEN) and 2-acetylaminofluorene (2AAF). A total of forty male BALB/c mice, weighing around 18 ± 2 g, were divided to four groups of ten. Group 1 was orally treated with a dose of corn oil, while Group 2 received a single intraperitoneal dose of DEN at 75 mg/kg, followed by 2AAF at 200 mg/kg dietary plan. Meanwhile, Group 3 was solely treated with 6-gingerol at 100 mg/kg. Lastly, Group 4 received 6-gingerol for 7 days prior to DEN + 2AAF exposure, with treatment maintained for 20 weeks.\u003c/p\u003e\n\u003cp\u003eResults: Administration of 6-gingerol conferred protective effects in HCC-induced mice by lowering the levels of TNF-α, IL-1β, COX-2, and iNOS. Its anti-inflammatory potential was further demonstrated through reduced expression of NF-κB and BCL2 in liver tissues, alongside increased levels of p53 and SIRT-1.\u003c/p\u003e\n\u003cp\u003eConclusion: The findings suggest that 6-gingerol counteracts DEN+2AAF-induced liver cancer by downregulating inflammatory mediators and promoting apoptosis through SIRT-1 activation.\u003c/p\u003e","manuscriptTitle":"Targeting early events in hepatocarcinogenesis: anti-inflammatory and SIRT1-mediated effects of 6-Gingerol in a murine model","msid":"","msnumber":"","nonDraftVersions":[{"code":2,"date":"2025-06-05 16:20:38","doi":"10.21203/rs.3.rs-6706491/v2","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}},{"code":1,"date":"2025-05-21 08:59:41","doi":"10.21203/rs.3.rs-6706491/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":"b7a25fc1-13d4-4772-8adf-003e53245af8","owner":[],"postedDate":"June 5th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":49478448,"name":"Cancer Biology"},{"id":49478449,"name":"Biochemical Research Methods"},{"id":49478450,"name":"Pathology"}],"tags":[],"updatedAt":"2025-05-21T08:59:41+00:00","versionOfRecord":[],"versionCreatedAt":"2025-06-05 16:20:38","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v2","identity":"rs-6706491","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6706491","identity":"rs-6706491","version":["v2"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}