Dihydroartemisinin attenuates inflammation and fibrosis in rats by suppressing JAK2/STAT3 signaling

Dihydroartemisinin attenuates inflammation and fibrosis in rats by suppressing JAK2/STAT3 signaling | 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 Dihydroartemisinin attenuates inflammation and fibrosis in rats by suppressing JAK2/STAT3 signaling Xiaolan You, xingyu jiang, Chuanmeng zhang, kejia jiang, xiaojun zhao, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-152531/v1 This work is licensed under a CC BY 4.0 License Status: Under Revision Version 1 posted 8 You are reading this latest preprint version Abstract Coronavirus disease 2019 (COVID-19), caused by SARS-CoV-2, has induced a worldwide pandemic since early 2020. COVID-19 causes pulmonary inflammation secondary to pulmonary fibrosis (PF); however, there are still no effective treatments for PF. The present study aimed to explore the inhibitory effect of dihydroartemisinin (DHA) on pulmonary inflammation and PF, and its molecular mechanism. Morphological changes and collagen deposition were analyzed using hematoxylin‑eosin staining, Masson staining, and the hydroxyproline content. DHA attenuated early alveolar inflammation and later PF in a bleomycin-induced rat PF model, and inhibited the expression of interleukin (IL) ‑1β, IL -6, tumor necrosis factor α (TNFα), and chemokine (C-C Motif) Ligand 3 (CCL3) in model rat serum. Further molecular analysis revealed that both pulmonary inflammation and PF were associated with increased transforming growth factor-β1 (TGF-β1), Janus activated kinase 2 (JAK2), and signal transducer and activator 3(STAT3) expression in the lung tissues of model rats. DHA reduces the inflammatory response and PF in the lungs by suppressing TGF-β1, JAK2, phosphorylated (p)-JAK2, STAT3, and p-STAT3 degradation. Thus, DHA exerts therapeutic effects against bleomycin-induced pulmonary inflammation and PF by inhibiting JAK2-STAT3 activation. Thus, DHA inhibits alveolar inflammation, and attenuates lung injury and fibrosis, possibly representing a therapeutic candidate to treat COVID-19. Drug Discovery, Design, & Development Infectious Diseases Coronavirus disease 2019 (COVID-19) pulmonary fibrosis dihydroartemisinin molecular mechanism Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Full Text Due to technical limitations, full-text HTML conversion of this manuscript could not be completed. However, the latest manuscript can be downloaded and accessed as a PDF. Additional Declarations No competing interests reported. Cite Share Download PDF Status: Under Revision Version 1 posted Editorial decision: Major revision 20 Apr, 2021 Reviews received at journal 20 Mar, 2021 Reviewers agreed at journal 08 Mar, 2021 Reviewers invited by journal 03 Mar, 2021 Editor assigned by journal 03 Mar, 2021 Editor invited by journal 15 Feb, 2021 Submission checks completed at journal 15 Feb, 2021 First submitted to journal 21 Jan, 2021 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-152531","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":12009809,"identity":"a89c1ccb-d514-47f7-8542-6cc5b0c75e69","order_by":0,"name":"Xiaolan You","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA0ElEQVRIiWNgGAWjYBACNmbm4x8+VNhAeDzEaOFnb0tjnHEmjQQtkj1n1Jh5Ww6ToMXgRg7bw5kN5xPnz0hgfPC2jUHenLCW3OMGH3fcTmyckcBsOLeNwXBnA0EteQmSM8/cTmyWSGCT5m1jSDA4QNhhBkCV5xLbJBLYfxOlBeh9M6CWA4k9QFuYidICDORkwxlnko1n8DxslpxzTsJwAyEtwKg8+OBDhZ3s/Pbkgx/elNnIE7QFBhwbGBgbgLQEkeqBwJ54paNgFIyCUTDiAABLk0Ru5rJ9BwAAAABJRU5ErkJggg==","orcid":"","institution":"Taizhou Clinical Medical School of Nanjing Medical University (Taizhou People’s Hospital)","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Xiaolan","middleName":"","lastName":"You","suffix":""},{"id":12009810,"identity":"018b29f0-6b77-4fdf-a946-6c42a66a6d9f","order_by":1,"name":"xingyu jiang","email":"","orcid":"","institution":"Department of Clinical Speciality, Nanjing Medical University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"xingyu","middleName":"","lastName":"jiang","suffix":""},{"id":12009811,"identity":"94c7e91f-0226-4964-91f8-c987c8d7d42e","order_by":2,"name":"Chuanmeng zhang","email":"","orcid":"","institution":"Taizhou Clinical Medical School of Nanjing Medical University (Taizhou People’s Hospital)","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Chuanmeng","middleName":"","lastName":"zhang","suffix":""},{"id":12009812,"identity":"08732348-2477-4ab4-995f-c4a3344d9e3c","order_by":3,"name":"kejia jiang","email":"","orcid":"","institution":"Taizhou Clinical Medical School of Nanjing Medical University (Taizhou People’s Hospital)","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"kejia","middleName":"","lastName":"jiang","suffix":""},{"id":12009813,"identity":"f5a44f46-169f-41a4-b121-8e2b2b5415cc","order_by":4,"name":"xiaojun zhao","email":"","orcid":"","institution":"Taizhou Clinical Medical School of Nanjing Medical University (Taizhou People’s Hospital)","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"xiaojun","middleName":"","lastName":"zhao","suffix":""},{"id":12009814,"identity":"8f80a0ab-e23c-45da-b0d1-84c3df4b3a45","order_by":5,"name":"ting guo","email":"","orcid":"","institution":"Taizhou Clinical Medical School of Nanjing Medical University (Taizhou People’s Hospital)","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"ting","middleName":"","lastName":"guo","suffix":""},{"id":12009815,"identity":"c3c896b0-2a81-4577-aea4-2bd0ce380e06","order_by":6,"name":"xiaowei zhu","email":"","orcid":"","institution":"Taizhou Clinical Medical School of Nanjing Medical University (Taizhou People’s Hospital)","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"xiaowei","middleName":"","lastName":"zhu","suffix":""},{"id":12009816,"identity":"0895f3f0-ed42-4471-8fda-5255ddb09835","order_by":7,"name":"jingjing Bao","email":"","orcid":"","institution":"Taizhou Clinical Medical School of Nanjing Medical University (Taizhou People’s Hospital)","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"jingjing","middleName":"","lastName":"Bao","suffix":""}],"badges":[],"createdAt":"2021-01-21 14:44:06","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-152531/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-152531/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":6034942,"identity":"a1fd479a-76a4-4e15-82c6-a179237bc944","added_by":"auto","created_at":"2021-02-16 23:58:31","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":1611608,"visible":true,"origin":"","legend":"DHA attenuates early alveolar inflammation and the inflammatory response in a bleomycin induced rat pulmonary fibrosis model . At 7 days after bleomycin intratracheal administration, with or without DHA treatment, the rats were sacrificed, and their lungs and blood were removed. (A) H\u0026E staining was used to detect the lung histopathological changes and (B) Lung inflammation was scored ( n = 6 ). (C) Il1b,(D) Il6, (E) Tnfa, (F) Ccl3 mRNA in lung tissues were determined to assess the lung inflammatory response (n = 6). Data are expressed as the means ± standard error . *P \u003c 0.05, **P \u003c 0.001. BLM, bleomycin; DHA, dihydroartemisinin; NS, not statistically significant.","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-152531/v1/ddf276e71c7b69fa22add338.png"},{"id":6034941,"identity":"8ca0b79a-cd69-48ed-a60f-79bee966d8f3","added_by":"auto","created_at":"2021-02-16 23:58:31","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":388716,"visible":true,"origin":"","legend":"DHA reduces the inflammatory response in the lungs via the JAK2/STAT3 signaling pathways. (A) IL-1β, (B) IL-6, (C) TNFα, and (D) CCL3 protein levels in blood were determined to assess the lung inflammatory response (n = 6). (E) Tgfb1, (F) Jak2, and (G) Stat3 mRNA levels in the lungs were determined using qRT-PCR (n = 6). Data are expressed as the means ± standard error . *P\u003c0.05, **P\u003c0.001. BLM, bleomycin; DHA, dihydroartemisinin; NS, not statistically significant; qRT-PCR, quantitative real-time reverse transcription PCR.","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-152531/v1/2f082cd82138b6ae1c2958e9.png"},{"id":6034812,"identity":"387481ec-3d98-4582-89ee-50aa9e3c67d1","added_by":"auto","created_at":"2021-02-16 23:55:31","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":2042102,"visible":true,"origin":"","legend":"DHA inhibited the JAK2/STAT3 pathway in pneumonia tissue. (A) Representative images of TGF β1, JAK2, p JAK2, STAT3, and p STAT3 protein expression in lung tissue detected by IHC. Mean IOD of (B) TGFβ1, (C) JAK2, (D)p-JAK2,(E) STAT3, and (F) p STAT3 protein by IHC staining. Data are expressed as the means ± standard error . *P \u003c 0.05, **P \u003c 0.001. BLM, bleomycin; DHA, dihydroartemisinin; IHC, immunohistochemistry; IOD, integrated optical density; NS, not statistically significant.","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-152531/v1/b27aa16ac05953ad54df8f28.png"},{"id":6034940,"identity":"242b75db-bda1-479e-8c98-c0506be4c402","added_by":"auto","created_at":"2021-02-16 23:58:31","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":1480309,"visible":true,"origin":"","legend":"DHA reduces pulmonary fibrosis induced by bleomycin in rats. Rats were\nsacrificed at 28 days after bleomycin injection, with or without DHA treatment, their\nblood and lungs were removed. (A) H\u0026E staining of bleomycin treated rat lungs, with\nand without DHA. (B) Lung inflammation was scored (n = 6). (C) IL-1β, (D) IL-6, (E) TNFα, (F) CCL3 protein in serum were determined by ELISA (n = 6). Data are expressed as the means ± standard error. *P \u003c 0.05, **P \u003c 0.001. BLM, bleomycin ; DHA, dihydroartemisinin; NS, not statistically significant; H\u0026E, hematoxylin and\neosin; ELISA, enzyme linked immunosorbent assay assay.","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-152531/v1/4f1cc4174bbfb06d215fa592.png"},{"id":6035297,"identity":"25ebab92-db68-4b46-8a14-8732be6775cd","added_by":"auto","created_at":"2021-02-17 00:01:31","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":1624567,"visible":true,"origin":"","legend":"DHA reduces collagen deposition in pulmonary fibrosis tissues through the\nJAK2/STAT3 pathway. The rats were sacrificed at 28 days after bleomycin injection,\nwith or without DHA treatment, and their lung were removed and blood collected. (A)\nRepresentative images of Masson staining of bleomycin treated rat lungs, with and\nwithout DHA. (B) Evaluation of pulmonary fibrosis u sing the Ashcroft score (n =\n6).(C) Determination of hydroxyproline in lung tissue(n = 6).(D) Tgfb1, Jak2, and (F) Stat3 mRNA levels in the lung tissues were determined using qRT-PCR (n = 6). Data are expressed as the means ± standard error . *P \u003c 0.05, **P \u003c 0.001. BLM, bleomycin; DHA, dihydroartemisinin; NS, not statistically significant; qRT-PCR, quantitative real-time reverse transcription PCR.","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-152531/v1/35a414fa4c0a5aff13fdbeae.png"},{"id":6034809,"identity":"f9865e83-7106-4921-8ae3-c7ce41897087","added_by":"auto","created_at":"2021-02-16 23:55:31","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":2016411,"visible":true,"origin":"","legend":"DHA inhibited pulmonary fibrosis via the JAK2/STAT3 pathway (A) Representative images of TGF β1, JAK2, p JAK2, STAT3, and p STAT3 protein in lung tissues, detected using IHC. Mean IOD of (B) TGF β1, (C) JAK2, (D) p JAK2,\n(E) STAT3, and (F) p STAT3 protein IHC staining. Data are expressed as the means ± standard error . *P \u003c 0.05, **P \u003c 0.001. BLM, bleomycin; DHA, dihydroartemisinin; IHC, immunohistochemistry; IOD, integrated optical d density. NS, not statistically significant.","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-152531/v1/3b129087bf814decb4eaf30d.png"},{"id":13660472,"identity":"968188c1-4924-43a2-8d37-1a70002d3b6d","added_by":"auto","created_at":"2021-09-17 10:24:45","extension":"pdf","order_by":6,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":3257460,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript215.pdf","url":"https://assets-eu.researchsquare.com/files/rs-152531/v1_covered.pdf"},{"id":13589285,"identity":"a9927863-b04c-4e33-8fb2-16ea416cdc00","added_by":"auto","created_at":"2021-09-17 04:59:48","extension":"pdf","order_by":6,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2374992,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript215.pdf","url":"https://assets-eu.researchsquare.com/files/rs-152531/v1_covered.pdf"},{"id":6035517,"identity":"91e8a9b1-f765-47a2-8730-2ff95755ef5d","added_by":"auto","created_at":"2021-02-17 00:04:36","extension":"pdf","order_by":6,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":3198832,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript215.pdf","url":"https://assets-eu.researchsquare.com/files/rs-152531/v1_stamped.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Dihydroartemisinin attenuates inflammation and fibrosis in rats by suppressing JAK2/STAT3 signaling","fulltext":[{"header":"Full Text","content":"Due to technical limitations, full-text HTML conversion of this manuscript could not be completed. However, the latest manuscript can be downloaded and \u003ca href='/article/rs-152531/latest.pdf' target='_blank'\u003e accessed as a PDF.\u003c/a\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Coronavirus disease 2019 (COVID-19), pulmonary fibrosis, dihydroartemisinin, molecular mechanism","lastPublishedDoi":"10.21203/rs.3.rs-152531/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-152531/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"Coronavirus disease 2019 (COVID-19), caused by SARS-CoV-2, has induced a worldwide pandemic since early 2020. COVID-19 causes pulmonary inflammation secondary to pulmonary fibrosis (PF); however, there are still no effective treatments for PF. The present study aimed to explore the inhibitory effect of dihydroartemisinin (DHA) on pulmonary inflammation and PF, and its molecular mechanism. Morphological changes and collagen deposition were analyzed using hematoxylin‑eosin staining, Masson staining, and the hydroxyproline content. DHA attenuated early alveolar inflammation and later PF in a bleomycin-induced rat PF model, and inhibited the expression of interleukin (IL) ‑1β, IL -6, tumor necrosis factor α (TNFα), and chemokine (C-C Motif) Ligand 3 (CCL3) in model rat serum. Further molecular analysis revealed that both pulmonary inflammation and PF were associated with increased transforming growth factor-β1 (TGF-β1), Janus activated kinase 2 (JAK2), and signal transducer and activator 3(STAT3) expression in the lung tissues of model rats. DHA reduces the inflammatory response and PF in the lungs by suppressing TGF-β1, JAK2, phosphorylated (p)-JAK2, STAT3, and p-STAT3 degradation. Thus, DHA exerts therapeutic effects against bleomycin-induced pulmonary inflammation and PF by inhibiting JAK2-STAT3 activation. Thus, DHA inhibits alveolar inflammation, and attenuates lung injury and fibrosis, possibly representing a therapeutic candidate to treat COVID-19.","manuscriptTitle":"Dihydroartemisinin attenuates inflammation and fibrosis in rats by suppressing JAK2/STAT3 signaling","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2021-02-16 23:55:29","doi":"10.21203/rs.3.rs-152531/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Major revision","date":"2021-04-20T11:59:34+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2021-03-20T19:26:12+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"fe413b89-b39f-46b6-90bc-9b2384c5ea9b","date":"2021-03-08T13:50:44+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2021-03-03T16:31:22+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2021-03-03T16:00:28+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2021-02-15T21:26:06+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2021-02-15T21:23:45+00:00","index":"","fulltext":""},{"type":"submitted","content":"Scientific Reports","date":"2021-01-21T14:31:01+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"1566c9e3-7654-45e1-8303-e4c2bc0e9a11","owner":[],"postedDate":"February 16th, 2021","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"in-revision","subjectAreas":[{"id":2455763,"name":"Drug Discovery, Design, \u0026 Development"},{"id":2455764,"name":"Infectious Diseases"}],"tags":[],"updatedAt":"2021-07-01T13:59:05+00:00","versionOfRecord":[],"versionCreatedAt":"2021-02-16 23:55:29","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-152531","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-152531","identity":"rs-152531","version":["v1"]},"buildId":"cBFmMYwuxLRRLfASyISRj","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}