Duodenal-jejunal bypass surgery activates eNOS and enhances antioxidant system by activating AMPK pathway to improve oxidative stress diabetic cardiomyopathy

Duodenal-jejunal bypass surgery activates eNOS and enhances antioxidant system by activating AMPK pathway to improve oxidative stress diabetic cardiomyopathy | 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 Duodenal-jejunal bypass surgery activates eNOS and enhances antioxidant system by activating AMPK pathway to improve oxidative stress diabetic cardiomyopathy guangwei yang, Xiang Zhao, Zitian Liu, Zheng Ge, Lunjin Yao, Zhiqiang Cheng, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-2452774/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: Diabetic cardiomyopathy (DCM) is a serious complication of Obesity with type 2 diabetes. Bariatric surgery can effectively improve DCM; however, the underlying mechanisms remain elusive. Our objective was to investigate the effect and potential mechanisms of duode-nal-jejunal bypass (DJB) on oxidative stress in the heart. Methods: DJB was performed on DCM rats, and high glucose (HG) and palmitate (PA) were used to simulate DCM in H9C2 cells in vitro. AMPK agonists and inhibitors were used to verify the role of the AMPK pathway. Serum from different groups of rats was used to verify that the im-provement in DCM after DJB surgery was not strictly owing to improvement in blood glucose levels and connect the experiments in vivo and in vitro. Results: DJB surgery effectively improved oxidative stress and increased expression of the AMPK pathway, p-eNOS/eNOS, NRF2, HO-1, and SOD2 in terms of protein and mRNA in the heart of DCM rats. Compared to H9C2 cells in the HG+PA environment alone, the addition of either AMPK agonists or serum from DJB rats increased the expression of the AMPK pathway, p-eNOS/eNOS, NRF2, HO-1, and SOD2 in terms of protein and mRNA and decreased the content of ROS, but this improvement was almost eliminated by the addition of AMPK inhibitors. Conclusions: Therefore, DJB surgery can improve oxidative stress in the heart of DCM rats, which may be achieved by activating the AMPK pathway, thereby increasing the phosphorylation level of eNOS, increasing NO and reducing ROS production, and improving the antioxidant system and not solely the improvement of blood glucose. Health sciences/Endocrinology/Endocrine system and metabolic diseases/Diabetes/Diabetes complications Health sciences/Diseases/Endocrine system and metabolic diseases/Diabetes/Diabetes complications Biological sciences/Physiology/Metabolism/Metabolic diseases/Diabetes/Diabetes complications duodenal-jejunal bypass diabetic cardiomyopathy oxidative stress eNOS AMPK an-31 tioxidative system Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Full Text Additional Declarations There is NO conflict of interest to disclose 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-2452774","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":167671213,"identity":"3e9ac5ed-035a-4664-b01c-d1fc390034b0","order_by":0,"name":"guangwei yang","email":"","orcid":"","institution":"","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"guangwei","middleName":"","lastName":"yang","suffix":""},{"id":167671214,"identity":"7dc42fc8-71af-471a-b21d-a251447cc9b3","order_by":1,"name":"Xiang Zhao","email":"","orcid":"","institution":"Shandong university","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Xiang","middleName":"","lastName":"Zhao","suffix":""},{"id":167671215,"identity":"6c7790ee-5160-4856-b852-fbb28da12358","order_by":2,"name":"Zitian Liu","email":"","orcid":"","institution":"Shandong University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Zitian","middleName":"","lastName":"Liu","suffix":""},{"id":167671216,"identity":"8ad2261b-c192-4a41-a6df-cf5a66f5a2db","order_by":3,"name":"Zheng Ge","email":"","orcid":"","institution":"Qilu Hospital of Shandong University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Zheng","middleName":"","lastName":"Ge","suffix":""},{"id":167671217,"identity":"e29dafea-123e-47d5-93d0-09f28b801196","order_by":4,"name":"Lunjin Yao","email":"","orcid":"","institution":"Shandong University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Lunjin","middleName":"","lastName":"Yao","suffix":""},{"id":167671218,"identity":"cf8cc217-5cff-409d-9f66-b87830b2d28d","order_by":5,"name":"Zhiqiang Cheng","email":"","orcid":"","institution":"Qilu Hospital of Shandong University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Zhiqiang","middleName":"","lastName":"Cheng","suffix":""},{"id":167671219,"identity":"88218ece-ab9f-4f1c-a968-deb82dfdf2b7","order_by":6,"name":"Xiaoyang Wang","email":"","orcid":"","institution":"Qilu Hospital of Shandong University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Xiaoyang","middleName":"","lastName":"Wang","suffix":""},{"id":167671220,"identity":"9e089d4e-c83f-45b4-8f86-3723a9f21aac","order_by":7,"name":"Xiang Zhang","email":"","orcid":"","institution":"Qilu Hospital of Shandong University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Xiang","middleName":"","lastName":"Zhang","suffix":""},{"id":167671221,"identity":"c16cab0c-3da2-4d7a-a78e-56e864bcaca9","order_by":8,"name":"Kexin Wang","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAvElEQVRIiWNgGAWjYDACHgbmx38qbOTY2NsPEK2FzYDnTJoxH8+ZBKK1MEjwth1KnCfhYECcDoMzZwwMJNgOpLdJMCQw/KjYRoSWs20JDwx47uS2STceYOw5c5uwFrPzzAcMEiSe5bbJHEhgZmwjSgtjg8QBg8PpbBIJBkRqOdt8QLIh4XAC8VrszxxLM2Y4kGbYBgzkg0T5RbInx/gx4z8befn29oMPflQQoQUFHCBR/SgYBaNgFIwCXAAArhM99EcQ72IAAAAASUVORK5CYII=","orcid":"","institution":"Qilu Hospital of Shandong University","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Kexin","middleName":"","lastName":"Wang","suffix":""}],"badges":[],"createdAt":"2023-01-07 08:35:40","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-2452774/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-2452774/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":31687187,"identity":"2836c508-e441-4586-97a9-1ca8176eff03","added_by":"auto","created_at":"2023-01-17 14:54:52","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":2073101,"visible":true,"origin":"","legend":"\u003cp\u003eDJB surgery improves general metabolic characteristics (A) The flow chart of animal experiments n=10 for per groups. (B). Body weight (C). Food intake (D). Fasting blood glucose (FBG) (E) curves of OGTT before surgery and (F).AUC OF OGTT before surgery (G) curves of OGTT 12 weeks after surgery (H) AUC of OGTT 12weeks after surgery. (I). HOMA-IR 12 weeks after surgery. Data are presented as mean ± SD. *P \u0026lt; 0.05, **P \u0026lt; 0.01 ***P\u0026lt; 0.001 CON vs. SHAM; #P \u0026lt; 0.05, ＃#P \u0026lt; 0.01 ###P \u0026lt; 0.001 DJB vs. SHAM. n = 10 in each group.\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-2452774/v1/f9b82815cb38787c2954cd8b.png"},{"id":31687193,"identity":"f967e72b-ed39-49c6-8579-50e00ad9d49a","added_by":"auto","created_at":"2023-01-17 14:54:56","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":20910000,"visible":true,"origin":"","legend":"\u003cp\u003eImprovement of DJB on DCM rats cardiac remodeling. (A). H\u0026amp;E staining images of left ventricle (B). \u0026nbsp;Masson staining of heart in rats of which dark blue indicated collagen fibers (C). the Sirus red staining to express collagen in rats’heart (D, E) Immunohistochemical staining of collagen 1 and colagen 3(F)WGA staining images and (G) Cardiomyocyte diameter are analyzed.***P\u0026lt; 0.001\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-2452774/v1/72d4e185957aab37bae7bfb9.png"},{"id":31687189,"identity":"d6def58c-d635-453e-955d-f3f125afaac1","added_by":"auto","created_at":"2023-01-17 14:54:52","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":5172600,"visible":true,"origin":"","legend":"\u003cp\u003eDJB surgery ameliorates cardiac AMPK/eNOS pathway and antioxidative system in DCM rats and improves oxidative stress in heart. (A) ROS fluorescence staining in the heart of three groups of rats and (B) the analysis results of relative ROS fluorescence intensity (C) Protein levels of antioxidative system related factors and pathway in \u0026nbsp;the heart of rats β-actin was an internal reference control. (D) Protein levels of AMPK pathway in the heart of rats, β-actin was an internal reference control（E）Relative mRNA levels of antioxidative system and Sirt1 in the heart of rats, \u0026nbsp;β-actin was a reference gene (F) Protein phosphorylation levels of eNOS (G）the content of NO in the heart muscle of rats 12 weeks after surgery. Data are presented as mean ± SD. *P \u0026lt; 0.05.**P \u0026lt; 0.01 ***P\u0026lt; 0.001, n=10 in each group.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-2452774/v1/1dfa0df66e89ecfb54a31589.png"},{"id":31688339,"identity":"80ef9665-6608-4aaf-b4c6-e0e801be44c9","added_by":"auto","created_at":"2023-01-17 15:10:52","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":2772076,"visible":true,"origin":"","legend":"\u003cp\u003eActivation of AMPK pathway can improve the eNOS activity and antioxidant system of H9C2 cell line \u0026nbsp;under HG+PA environment. (A) ROS staining in H9C2 cells, shsown by green fluorescence. (B) Protein levels of anti-oxidative system were analyzed in H9C2 cells ,β-actin was an internal reference control (C) Relative mRNA levels of anti-oxidation system and Sirt1 genes in rats .β-actin served as a reference gene (D) Protein levels of AMPK pathway \u0026nbsp;in H9C2 cells, β-actin was an internal reference (E) Protein phosphorylation levels of eNOS In H9C2 cells, β-actin was an internal reference (F). Content of NO in H9C2 cells Data are presented as mean ± SD. *P \u0026lt; 0.05.**P \u0026lt; 0.01 ***P\u0026lt; 0.001 n=3 in each group.\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-2452774/v1/62bbe4ba0a012f1181c5d082.png"},{"id":31687191,"identity":"c5466964-dac6-44b3-b2ef-d8e4d1767e7f","added_by":"auto","created_at":"2023-01-17 14:54:52","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":3445736,"visible":true,"origin":"","legend":"\u003cp\u003eEffects of CON.SHAM and DJB group rat serum on AMPK/eNOS pathway and antioxidant system of H9C2 cells in HG+PA environment(A) ROS staining in H9C2 cells, shsown by green fluorescence. (B) Protein levels of \u0026nbsp;AMPK pathway in H9C2 cells, β-actin was an internal reference (C) Relative mRNA levels of anti-oxidation system and Sirt1 genes in rats , β-actin served as a reference gene (D)Protein levels of antioxidative system in H9C2 cells ,β-actin was an internal reference control (E) Protein phosphorylation levels of eNOS In H9C2 cells ,β-actin was an internal reference (F). Content of NO in H9C2 cells Data are presented as mean ± SD. *P \u0026lt; 0.05.**P \u0026lt; 0.01 ***P\u0026lt; 0.001 n=3 in each group.\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-2452774/v1/a0bd135eb162cedf6e0192d0.png"},{"id":31688026,"identity":"dc14699a-f116-4c83-9101-7355d93b2d93","added_by":"auto","created_at":"2023-01-17 15:02:52","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":2157845,"visible":true,"origin":"","legend":"\u003cp\u003eImprovement of oxidative stress of H9C2 cells in HG+PA environment by serum of DJB rats depends on the activation of AMPK pathway to a certain extent(A) ROS staining in H9C2 cells, shown by green fluorescence. (B) Relative mRNA levels of anti-oxidation system genes in rats. β-actin served as a reference gene(C) Ｐrotein levels of antioxidative system in H9C2 cells ,β-actin was an internal reference control (D) Protein levels of AMPK pathway in H9C2 cells ,β-actin was an internal reference (E) Protein phosphorylation levels of eNOS In H9C2 cells ,β-actin was an internal reference (F). Content of NO in H9C2 cells Data are presented as mean ± SD. *P \u0026lt; 0.05.**P \u0026lt; 0.01 ***P\u0026lt; 0.001 n=3 in each group.\u003c/p\u003e","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-2452774/v1/d2455f3a42f2e9383a2b3fc4.png"},{"id":39297902,"identity":"627c8aa2-01b6-4e83-bdba-90b12bd994f4","added_by":"auto","created_at":"2023-06-29 13:34:52","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1147350,"visible":true,"origin":"","legend":"","description":"","filename":"eNOSsubmit2.pdf","url":"https://assets-eu.researchsquare.com/files/rs-2452774/v1_covered_35501162-a89a-4829-8391-5f7e95c4efbf.pdf"}],"financialInterests":"There is \u003cb\u003eNO\u003c/b\u003e conflict of interest to disclose","formattedTitle":"Duodenal-jejunal bypass surgery activates eNOS and enhances antioxidant system by activating AMPK pathway to improve oxidative stress diabetic cardiomyopathy","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":"duodenal-jejunal bypass, diabetic cardiomyopathy, oxidative stress, eNOS, AMPK, an-31 tioxidative system","lastPublishedDoi":"10.21203/rs.3.rs-2452774/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-2452774/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground:\u003c/strong\u003e Diabetic cardiomyopathy (DCM) is a serious complication of Obesity with type 2 diabetes. Bariatric surgery can effectively improve DCM; however, the underlying mechanisms remain elusive. Our objective was to investigate the effect and potential mechanisms of duode-nal-jejunal bypass (DJB) on oxidative stress in the heart.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods:\u003c/strong\u003e DJB was performed on DCM rats, and high glucose (HG) and palmitate (PA) were used to simulate DCM in H9C2 cells in vitro. AMPK agonists and inhibitors were used to verify the role of the AMPK pathway. Serum from different groups of rats was used to verify that the im-provement in DCM after DJB surgery was not strictly owing to improvement in blood glucose levels and connect the experiments in vivo and in vitro.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults:\u003c/strong\u003e DJB surgery effectively improved oxidative stress and increased expression of the AMPK pathway, p-eNOS/eNOS, NRF2, HO-1, and SOD2 in terms of protein and mRNA in the heart of DCM rats. Compared to H9C2 cells in the HG+PA environment alone, the addition of either AMPK agonists or serum from DJB rats increased the expression of the AMPK pathway, p-eNOS/eNOS, NRF2, HO-1, and SOD2 in terms of protein and mRNA and decreased the content of ROS, but this improvement was almost eliminated by the addition of AMPK inhibitors.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusions: \u003c/strong\u003eTherefore, DJB surgery can improve oxidative stress in the heart of DCM rats, which may be achieved by activating the AMPK pathway, thereby increasing the phosphorylation level of eNOS, increasing NO and reducing ROS production, and improving the antioxidant system and not solely the improvement of blood glucose.\u003c/p\u003e","manuscriptTitle":"Duodenal-jejunal bypass surgery activates eNOS and enhances antioxidant system by activating AMPK pathway to improve oxidative stress diabetic cardiomyopathy","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2023-01-17 14:54:47","doi":"10.21203/rs.3.rs-2452774/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":"aaca06be-f47f-43cb-aec3-788773f9d7d9","owner":[],"postedDate":"January 17th, 2023","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":18405142,"name":"Health sciences/Endocrinology/Endocrine system and metabolic diseases/Diabetes/Diabetes complications"},{"id":18405143,"name":"Health sciences/Diseases/Endocrine system and metabolic diseases/Diabetes/Diabetes complications"},{"id":18405144,"name":"Biological sciences/Physiology/Metabolism/Metabolic diseases/Diabetes/Diabetes complications"}],"tags":[],"updatedAt":"2023-06-29T13:26:35+00:00","versionOfRecord":[],"versionCreatedAt":"2023-01-17 14:54:47","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-2452774","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-2452774","identity":"rs-2452774","version":["v1"]},"buildId":"WrCJVZZCHTDjtuVLN7oU0","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}