{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,15]],"date-time":"2026-10-15T08:28:47Z","timestamp":1792052927763,"version":"4.4.0"},"reference-count":27,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2023,11,29]],"date-time":"2023-11-29T00:00:00Z","timestamp":1701216000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["82171336"],"award-info":[{"award-number":["82171336"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["81870939"],"award-info":[{"award-number":["81870939"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Brain Sciences"],"abstract":"<jats:p>Background and Aims: The immune-inflammatory cascade and pyroptosis play an important role in the pathogenesis of cerebral ischemia-reperfusion injury (CIRI). The maintenance of immune homeostasis is inextricably linked to the Notch signaling pathway, but whether myeloid Notch1 affects microglia polarization as well as neuronal pyroptosis in CIRI is not fully understood. This study was designed to clarify the role of myeloid Notch1 in CIRI, providing new therapeutic strategies for ischemic stroke. Methods and Results: Myeloid-specific Notch1 knockout (Notch1M-KO) mice and the floxed Notch1 (Notch1FL\/FL) mice were subjected to middle cerebral artery occlusion (MCAO). After 3 days of CIRI, we evaluated the neurological deficit score and cerebral infarction volume. Immunofluorescence staining was used to detect the expression of Notch1 and microglial subtype markers. Cerebral infiltrating macrophages were detected by flow cytometry. RT-qPCR was used to detect pro-inflammatory cytokines. Western blot was used to detect the expression of pyroptosis related proteins. The Notch1-siRNA transfected BV2 cells were co-cultured with HT22 cells to investigate the potential mechanisms by which microglial Notch1 affects neuronal pyroptosis induced by anoxia\/reoxygenation in vitro. We found that Notch1 was activated in cerebral microglia\/macrophages after CIRI. Myeloid Notch1 deficiency decreased the cerebral infarct volume (24.17 \u00b1 3.29 vs. 36.17 \u00b1 2.27, p &lt; 0.001), neurological function scores (2.33 \u00b1 0.47 vs. 3.17 \u00b1 0.37, p &lt; 0.001) and the infiltration of peripheral monocytes\/macrophages (3.26 \u00b1 0.53 vs. 5.67 \u00b1 0.57, p &lt; 0.01). Strikingly, myeloid-specific Notch1 knockout alleviated pyroptosis. Compared with microglia M1, increased microglia M2 were detected in the ischemic penumbra. In parallel in vitro co-culture experiments, we found that Notch1 knockdown in microglial BV2 cells inhibited anoxia\/reoxygenation-induced JAK2\/STAT3 activation and pyroptosis in hippocampal neuron HT22 cells. Conclusions: Our findings elucidate the underlying mechanism of the myeloid Notch1 signaling pathway in regulating neuronal pyroptosis in CIRI, suggesting that targeting myeloid-specific Notch1 is an effective strategy for the treatment of ischemic stroke.<\/jats:p>","DOI":"10.3390\/brainsci13121657","type":"journal-article","created":{"date-parts":[[2023,11,29]],"date-time":"2023-11-29T09:00:18Z","timestamp":1701248418000},"page":"1657","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":16,"title":["Targeting Microglia\/Macrophages Notch1 Protects Neurons from Pyroptosis in Ischemic Stroke"],"prefix":"10.3390","volume":"13","author":[{"given":"Ran","family":"Chen","sequence":"first","affiliation":[{"name":"Department of Neurosurgery, Renmin Hospital of Wuhan University, Wuhan 430064, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hua","family":"Zhu","sequence":"additional","affiliation":[{"name":"Department of Neurosurgery, Renmin Hospital of Wuhan University, Wuhan 430064, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhihui","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Neurosurgery, Renmin Hospital of Wuhan University, Wuhan 430064, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yonggang","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Neurosurgery, Renmin Hospital of Wuhan University, Wuhan 430064, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jin","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Anesthesia, Renmin Hospital of Wuhan University, Wuhan 430064, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yingao","family":"Huang","sequence":"additional","affiliation":[{"name":"Department of Neurosurgery, Renmin Hospital of Wuhan University, Wuhan 430064, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lijuan","family":"Gu","sequence":"additional","affiliation":[{"name":"Central Laboratory, Renmin Hospital of Wuhan University, Wuhan 430064, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4001-037X","authenticated-orcid":false,"given":"Changyong","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Physiology, School of Basic Medical Sciences, Wuhan University, Wuhan 430072, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6983-8547","authenticated-orcid":false,"given":"Xiaoxing","family":"Xiong","sequence":"additional","affiliation":[{"name":"Department of Neurosurgery, Renmin Hospital of Wuhan University, Wuhan 430064, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhihong","family":"Jian","sequence":"additional","affiliation":[{"name":"Department of Neurosurgery, Renmin Hospital of Wuhan University, Wuhan 430064, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,11,29]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"307","DOI":"10.1007\/s12975-016-0518-y","article-title":"The high cost of stroke and stroke cytoprotection research","volume":"8","author":"Lapchak","year":"2017","journal-title":"Transl. Stroke Res."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1016\/bs.pbr.2015.01.003","article-title":"Stroke rehabilitation: Clinical picture, assessment, and therapeutic challenge","volume":"218","author":"Richards","year":"2015","journal-title":"Prog. Brain Res."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Lv, S., Liu, H., and Wang, H. (2021). The interplay between autophagy and nlrp3 inflammasome in ischemia\/reperfusion injury. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22168773"},{"key":"ref_4","first-page":"1435","article-title":"The new language of cerebral ischemia","volume":"18","author":"Ginsberg","year":"1997","journal-title":"AJNR Am. J. Neuroradiol."},{"key":"ref_5","first-page":"965","article-title":"Therapeutic effects of herbal compounds in cerebral ischemia with special reference to suppression of microglia activation implicated in neurodegeneration","volume":"34","author":"Jia","year":"2019","journal-title":"Histol. Histopathol."},{"key":"ref_6","first-page":"2006","article-title":"Modulators of microglia activation and polarization in ischemic stroke (review)","volume":"21","author":"Jiang","year":"2020","journal-title":"Mol. Med. Rep."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"105419","DOI":"10.1016\/j.phrs.2021.105419","article-title":"Phytochemicals as regulators of microglia\/macrophages activation in cerebral ischemia","volume":"165","author":"Subedi","year":"2021","journal-title":"Pharmacol. Res."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"3083","DOI":"10.2147\/JIR.S350109","article-title":"Microglia-mediated neuroinflammation: A potential target for the treatment of cardiovascular diseases","volume":"15","author":"Wang","year":"2022","journal-title":"J. Inflamm. Res."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"26162","DOI":"10.1038\/srep26162","article-title":"Endocytosis of indium-tin-oxide nanoparticles by macrophages provokes pyroptosis requiring nlrp3-asc-caspase1 axis that can be prevented by mesenchymal stem cells","volume":"6","author":"Naji","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1038\/nrmicro2070","article-title":"Pyroptosis: Host cell death and inflammation","volume":"7","author":"Bergsbaken","year":"2009","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_11","first-page":"5685","article-title":"Inhibition of hepatic cells pyroptosis attenuates CLP-induced acute liver injury","volume":"8","author":"Chen","year":"2016","journal-title":"Am. J. Transl. Res."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"329","DOI":"10.1128\/EC.00336-13","article-title":"Candida albicans triggers nlrp3-mediated pyroptosis in macrophages","volume":"13","author":"Wellington","year":"2014","journal-title":"Eukaryot. Cell"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"270","DOI":"10.1186\/s12974-020-01946-7","article-title":"Cottonseed oil alleviates ischemic stroke injury by inhibiting the inflammatory activation of microglia and astrocyte","volume":"17","author":"Liu","year":"2020","journal-title":"J. Neuroinflamm."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1016\/j.tibs.2016.10.004","article-title":"Pyroptosis: Gasdermin-mediated programmed necrotic cell death","volume":"42","author":"Shi","year":"2017","journal-title":"Trends Biochem. Sci."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Wellington, M., Koselny, K., and Krysan, D.J. (2012). Candida albicans morphogenesis is not required for macrophage interleukin 1beta production. mBio, 4.","DOI":"10.1128\/mBio.00433-12"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1353","DOI":"10.4049\/jimmunol.1600699","article-title":"Active caspase-1 induces plasma membrane pores that precede pyroptotic lysis and are blocked by lanthanides","volume":"197","author":"Russo","year":"2016","journal-title":"J. Immunol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1471","DOI":"10.1111\/j.1471-4159.2008.05715.x","article-title":"Notch: From neural development to neurological disorders","volume":"107","author":"Lathia","year":"2008","journal-title":"J. Neurochem."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"621","DOI":"10.1038\/nm1403","article-title":"Gamma secretase-mediated notch signaling worsens brain damage and functional outcome in ischemic stroke","volume":"12","author":"Arumugam","year":"2006","journal-title":"Nat. Med."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"714943","DOI":"10.3389\/fimmu.2021.714943","article-title":"Janus kinase inhibition ameliorates ischemic stroke injury and neuroinflammation through reducing nlrp3 inflammasome activation via jak2\/stat3 pathway inhibition","volume":"12","author":"Zhu","year":"2021","journal-title":"Front. Immunol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"D362","DOI":"10.1093\/nar\/gkw937","article-title":"The string database in 2017: Quality-controlled protein-protein association networks, made broadly accessible","volume":"45","author":"Szklarczyk","year":"2017","journal-title":"Nucleic Acids Res."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"996","DOI":"10.1097\/ALN.0b013e31826cb469","article-title":"Activation of canonical notch signaling pathway is involved in the ischemic tolerance induced by sevoflurane preconditioning in mice","volume":"117","author":"Yang","year":"2012","journal-title":"Anesthesiology"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"4795","DOI":"10.1111\/jcmm.14368","article-title":"Folic acid deficiency enhanced microglial immune response via the notch1\/nuclear factor kappa B p65 pathway in hippocampus following rat brain I\/R injury and BV2 cells","volume":"23","author":"Cheng","year":"2019","journal-title":"J. Cell Mol. Med."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1016\/j.biomaterials.2018.01.039","article-title":"Protecting neurons from cerebral ischemia\/reperfusion injury via nanoparticle-mediated delivery of an sirna to inhibit microglial neurotoxicity","volume":"161","author":"Wang","year":"2018","journal-title":"Biomaterials"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"242","DOI":"10.1186\/s12974-018-1282-6","article-title":"Mitochondrial dysfunction induces nlrp3 inflammasome activation during cerebral ischemia\/reperfusion injury","volume":"15","author":"Gong","year":"2018","journal-title":"J. Neuroinflamm."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"6424","DOI":"10.7150\/thno.35528","article-title":"Targeted inhibition of stat3 as a potential treatment strategy for atherosclerosis","volume":"9","author":"Chen","year":"2019","journal-title":"Theranostics"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"64","DOI":"10.3389\/fphar.2020.00064","article-title":"Protective effect of stachydrine against cerebral ischemia-reperfusion injury by reducing inflammation and apoptosis through p65 and jak2\/stat3 signaling pathway","volume":"11","author":"Li","year":"2020","journal-title":"Front. Pharmacol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"5638943","DOI":"10.1155\/2016\/5638943","article-title":"Notch1 pathway protects against burn-induced myocardial injury by repressing reactive oxygen species production through jak2\/stat3 signaling","volume":"2016","author":"Cai","year":"2016","journal-title":"Oxid. Med. Cell. Longev."}],"container-title":["Brain Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-3425\/13\/12\/1657\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T21:33:52Z","timestamp":1760132032000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-3425\/13\/12\/1657"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,11,29]]},"references-count":27,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2023,12]]}},"alternative-id":["brainsci13121657"],"URL":"https:\/\/doi.org\/10.3390\/brainsci13121657","relation":{},"ISSN":["2076-3425"],"issn-type":[{"value":"2076-3425","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,11,29]]}}}