Formononetin reverses immune-mediated TregTh17 immune imbalance in Immune-mediated bone marrow failure mice by regulating the PI3KAkt signaling pathway

Formononetin reverses immune-mediated TregTh17 immune imbalance in Immune-mediated bone marrow failure mice by regulating the PI3KAkt signaling 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 Research Article Formononetin reverses immune-mediated TregTh17 immune imbalance in Immune-mediated bone marrow failure mice by regulating the PI3KAkt signaling pathway hui xuan lan, Wei Qiu, Jie Wu, Zhijing Hu, Xiaomin Zhang, lingling zhu This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-3463325/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 25 Mar, 2024 Read the published version in Chinese Medicine → Version 1 posted 5 You are reading this latest preprint version Abstract Background: Severe aplastic anemia (SAA) is a syndrome of bone marrow failure which is life-threatening. Recent research has discovered that CD4+ T cell subsets, including Treg cells and Th17 cells, play a pivotal role in the pathogenesis of SAA. Formononetin (FMN) is a natural compound extracted from the traditional Chinese medicine Huangqi, which has the ability to regulate the imbalance of Treg/Th17 cells in some inflammatory diseases. Nevertheless, the therapeutic effect of FMN in SAA has yet to be definitively established. Therefore, the objective of this research was to investigate the effect of FMN on SAA and elucidate its underlying mechanism. Methods: In vivo experiments, the mice were divided into five groups: the control group, model group, low- and high- dose of FMN groups and positive control cyclosporine A group. The immune-mediated bone marrow failure (BMF) mouse model was established by the total body X-ray radiation and lymphocyte infusion. After 10 days of continuous administration of FMN, the numbers of Treg/Th17 cells in the bone marrow and spleen were assessed by flow cytometry. The protein expressions of PI3K/Akt pathway in the bone marrow and spleen was assessed by immunohistochemistry and western blotting. In vitro, the impact of FMN on the differentiation of naive CD4+T cells into Treg cells was investigated by flow cytometry and ELISA. Results: In comparison with the control group, the model group showed a reduction in bone marrow nucleated cells, a significant decrease in peripheral blood cells, and an altered CD8+/CD4+T cell ratio. These findings indicate the successful establishment of a mouse model of immune-mediated BMF. After FMN treatment, there were the increased levels of red blood cells, hemoglobin and platelets. In addition, FMN mitigated the bone marrow destruction and restored the CD8+/CD4+ T cell ratio. Furthermore, in comparison with the control group, the model group showed the decreased levels of Treg cells and the increased levels of Th17 cells. After FMN treatment, there was a significantly increased number of Treg cells and a decreased number of Th17 cells. Additionally, FMN remarkably down-regulated the expression levels of PI3K and Akt proteins in immune-mediated BMF mice. Conclusions: FMN alleviates immune-mediated BMF by modulating the balance of Treg/Th17 cells through the PI3K/Akt signaling pathway. Formononetin Treg/Th17 cells Severe aplastic anemia PI3K/Akt signaling pathway Full Text Cite Share Download PDF Status: Published Journal Publication published 25 Mar, 2024 Read the published version in Chinese Medicine → Version 1 posted Editorial decision: Major revision 07 Dec, 2023 Reviewers agreed at journal 13 Nov, 2023 Reviewers invited by journal 02 Nov, 2023 Editor assigned by journal 01 Nov, 2023 First submitted to journal 21 Oct, 2023 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. 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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-3463325","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":245460521,"identity":"c2fc6eb8-4b58-4719-a1b3-8d0d7508093b","order_by":0,"name":"hui xuan lan","email":"","orcid":"","institution":"Southern Medical University","correspondingAuthor":false,"prefix":"","firstName":"hui","middleName":"xuan","lastName":"lan","suffix":""},{"id":245460522,"identity":"5dfb8de1-d6e6-45f4-b84d-8f6b25b3cd91","order_by":1,"name":"Wei Qiu","email":"","orcid":"","institution":"Southern Medical 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Recent research has discovered that CD4+ T cell subsets, including Treg cells and Th17 cells, play a pivotal role in the pathogenesis of SAA. Formononetin (FMN) is a natural compound extracted from the traditional Chinese medicine Huangqi, which has the ability to regulate the imbalance of Treg/Th17 cells in some inflammatory diseases. Nevertheless, the therapeutic effect of FMN in SAA has yet to be definitively established. Therefore, the objective of this research was to investigate the effect of FMN on SAA and elucidate its underlying mechanism.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eMethods: In vivo experiments, the mice were divided into five groups: the control group, model group, low- and high- dose of FMN groups and positive control cyclosporine A group. The immune-mediated bone marrow failure (BMF) mouse model was established by the total body X-ray radiation and lymphocyte infusion. After 10 days of continuous administration of FMN, the numbers of Treg/Th17 cells in the bone marrow and spleen were assessed by flow cytometry. The protein expressions of PI3K/Akt pathway in the bone marrow and spleen was assessed by immunohistochemistry and western blotting. In vitro, the impact of FMN on the differentiation of naive CD4+T cells into Treg cells was investigated by flow cytometry and ELISA.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eResults: In comparison with the control group, the model group showed a reduction in bone marrow nucleated cells, a significant decrease in peripheral blood cells, and an altered CD8+/CD4+T cell ratio. These findings indicate the successful establishment of a mouse model of immune-mediated BMF. After FMN treatment, there were the increased levels of red blood cells, hemoglobin and platelets. In addition, FMN mitigated the bone marrow destruction and restored the CD8+/CD4+ T cell ratio. 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Additionally, FMN remarkably down-regulated the expression levels of PI3K and Akt proteins in immune-mediated BMF mice.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eConclusions: FMN alleviates immune-mediated BMF by modulating the balance of Treg/Th17 cells through the PI3K/Akt signaling pathway.\u003c/p\u003e","manuscriptTitle":"Formononetin reverses immune-mediated TregTh17 immune imbalance in Immune-mediated bone marrow failure mice by regulating the PI3KAkt signaling pathway","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2023-11-07 15:55:59","doi":"10.21203/rs.3.rs-3463325/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Major revision","date":"2023-12-07T05:50:30+00:00","index":"","fulltext":""},{"type":"reviewerAgreed","content":"","date":"2023-11-13T06:50:04+00:00","index":0,"fulltext":""},{"type":"reviewersInvited","content":"","date":"2023-11-03T03:45:47+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2023-11-02T03:58:31+00:00","index":"","fulltext":""},{"type":"submitted","content":"Chinese Medicine","date":"2023-10-22T03:10:20+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"chinese-medicine","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"cmed","sideBox":"Learn more about [Chinese Medicine](http://cmjournal.biomedcentral.com)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/cmed/default.aspx","title":"Chinese Medicine","twitterHandle":"@BioMedCentral","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"1d5f959d-234e-438c-9c8b-5610f56f27bd","owner":[],"postedDate":"November 7th, 2023","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2024-04-01T15:04:38+00:00","versionOfRecord":{"articleIdentity":"rs-3463325","link":"https://doi.org/10.1186/s13020-024-00919-9","journal":{"identity":"chinese-medicine","isVorOnly":false,"title":"Chinese Medicine"},"publishedOn":"2024-03-25 15:00:36","publishedOnDateReadable":"March 25th, 2024"},"versionCreatedAt":"2023-11-07 15:55:59","video":"","vorDoi":"10.1186/s13020-024-00919-9","vorDoiUrl":"https://doi.org/10.1186/s13020-024-00919-9","workflowStages":[]},"version":"v1","identity":"rs-3463325","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-3463325","identity":"rs-3463325","version":["v1"]},"buildId":"J0_U0BvcaRcwD8yVFaRlm","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}