{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,4]],"date-time":"2026-09-04T17:49:15Z","timestamp":1788544155798,"version":"build-2803163510"},"reference-count":40,"publisher":"MDPI AG","issue":"13","license":[{"start":{"date-parts":[[2022,6,25]],"date-time":"2022-06-25T00:00:00Z","timestamp":1656115200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Interdisciplinary Center for Clinical Research","award":["Z-3\/66"],"award-info":[{"award-number":["Z-3\/66"]}]},{"name":"Interdisciplinary Center for Clinical Research","award":["374031971\u2013CRC\/TR 240"],"award-info":[{"award-number":["374031971\u2013CRC\/TR 240"]}]},{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["Z-3\/66"],"award-info":[{"award-number":["Z-3\/66"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["374031971\u2013CRC\/TR 240"],"award-info":[{"award-number":["374031971\u2013CRC\/TR 240"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Open Access Publication Fund of the University of Wuerzburg","award":["Z-3\/66"],"award-info":[{"award-number":["Z-3\/66"]}]},{"name":"Open Access Publication Fund of the University of Wuerzburg","award":["374031971\u2013CRC\/TR 240"],"award-info":[{"award-number":["374031971\u2013CRC\/TR 240"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>In large vessel occlusion stroke, recanalization to restore cerebral perfusion is essential but not necessarily sufficient for a favorable outcome. Paradoxically, in some patients, reperfusion carries the risk of increased tissue damage and cerebral hemorrhage. Experimental and clinical data suggest that endothelial cells, representing the interface for detrimental platelet and leukocyte responses, likely play a crucial role in the phenomenon referred to as ischemia\/reperfusion (I\/R)-injury, but the mechanisms are unknown. We aimed to determine the role of endoglin in cerebral I\/R-injury; endoglin is a membrane-bound protein abundantly expressed by endothelial cells that has previously been shown to be involved in the maintenance of vascular homeostasis. We investigated the expression of membranous endoglin (using Western blotting and RT-PCR) and the generation of soluble endoglin (using an enzyme-linked immunosorbent assay of cell culture supernatants) after hypoxia and subsequent reoxygenation in human non-immortalized brain endothelial cells. To validate these in vitro data, we additionally examined endoglin expression in an intraluminal monofilament model of permanent and transient middle cerebral artery occlusion in mice. Subsequently, the effects of recombinant human soluble endoglin were assessed by label-free impedance-based measurement of endothelial monolayer integrity (using the xCELLigence DP system) and immunocytochemistry. Endoglin expression is highly inducible by hypoxia in human brain endothelial monolayers in vitro, and subsequent reoxygenation induced its shedding. These findings were corroborated in mice during MCAO; an upregulation of endoglin was displayed in the infarcted hemispheres under occlusion, whereas endoglin expression was significantly diminished after transient MCAO, which is indicative of shedding. Of note is the finding that soluble endoglin induced an inflammatory phenotype in endothelial monolayers. The treatment of HBMEC with endoglin resulted in a decrease in transendothelial resistance and the downregulation of VE-cadherin. Our data establish a novel mechanism in which hypoxia triggers the initial endothelial upregulation of endoglin and subsequent reoxygenation triggers its release as a vasoactive mediator that, when rinsed into adjacent vascular beds after recanalization, can contribute to cerebral reperfusion injury.<\/jats:p>","DOI":"10.3390\/ijms23137085","type":"journal-article","created":{"date-parts":[[2022,6,26]],"date-time":"2022-06-26T09:00:13Z","timestamp":1656234013000},"page":"7085","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Regulation and Release of Vasoactive Endoglin by Brain Endothelium in Response to Hypoxia\/Reoxygenation in Stroke"],"prefix":"10.3390","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5597-6225","authenticated-orcid":false,"given":"Axel","family":"Haarmann","sequence":"first","affiliation":[{"name":"Department of Neurology, University of W\u00fcrzburg, 97080 W\u00fcrzburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4350-3655","authenticated-orcid":false,"given":"Lena","family":"Zimmermann","sequence":"additional","affiliation":[{"name":"Department of Neurology, University of W\u00fcrzburg, 97080 W\u00fcrzburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5535-6480","authenticated-orcid":false,"given":"Michael","family":"Bieber","sequence":"additional","affiliation":[{"name":"Department of Neurology, University of W\u00fcrzburg, 97080 W\u00fcrzburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christine","family":"Silwedel","sequence":"additional","affiliation":[{"name":"University Children\u2019s Hospital, University of W\u00fcrzburg, 97080 W\u00fcrzburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guido","family":"Stoll","sequence":"additional","affiliation":[{"name":"Department of Neurology, University of W\u00fcrzburg, 97080 W\u00fcrzburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael K.","family":"Schuhmann","sequence":"additional","affiliation":[{"name":"Department of Neurology, University of W\u00fcrzburg, 97080 W\u00fcrzburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,6,25]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2760","DOI":"10.1161\/STROKEAHA.117.016456","article-title":"Systematic Evaluation of Patients Treated With Neurothrombectomy Devices for Acute Ischemic Stroke: Primary Results of the STRATIS Registry","volume":"48","author":"Zaidat","year":"2017","journal-title":"Stroke"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"603","DOI":"10.1016\/j.neuron.2014.03.003","article-title":"Stepwise recruitment of transcellular and paracellular pathways underlies blood-brain barrier breakdown in stroke","volume":"82","author":"Knowland","year":"2014","journal-title":"Neuron"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"126","DOI":"10.1016\/j.jneumeth.2009.03.002","article-title":"Near-infrared fluorescence imaging with fluorescently labeled albumin: A novel method for non-invasive optical imaging of blood-brain barrier impairment after focal cerebral ischemia in mice","volume":"180","author":"Klohs","year":"2009","journal-title":"J. Neurosci. Methods"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1038\/jcbfm.2013.203","article-title":"Hemorrhagic transformation after ischemic stroke in animals and humans","volume":"34","author":"Jickling","year":"2014","journal-title":"J. Cereb. Blood Flow Metab."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1269","DOI":"10.1007\/s00018-017-2694-7","article-title":"Human endoglin as a potential new partner involved in platelet-endothelium interactions","volume":"75","author":"Rossi","year":"2018","journal-title":"Cell. Mol. Life Sci."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"2653","DOI":"10.1161\/01.STR.31.11.2653","article-title":"Analysis of endoglin expression in normal brain tissue and in cerebral arteriovenous malformations","volume":"31","author":"Matsubara","year":"2000","journal-title":"Stroke"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"2677","DOI":"10.1242\/jcs.00470","article-title":"CD105 prevents apoptosis in hypoxic endothelial cells","volume":"116","author":"Li","year":"2003","journal-title":"J. Cell Sci."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"283","DOI":"10.1016\/j.bbrc.2009.12.170","article-title":"Endothelial cells are activated during hypoxia via endoglin\/ALK-1\/SMAD1\/5 signaling in vivo and in vitro","volume":"392","author":"Tian","year":"2010","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"199","DOI":"10.20517\/2347-8659.2017.18","article-title":"The roles of endoglin gene in cerebrovascular diseases","volume":"4","author":"Zhu","year":"2017","journal-title":"Neuroimmunol. Neuroinflamm."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"345","DOI":"10.1038\/ng1294-345","article-title":"Endoglin, a TGF-beta binding protein of endothelial cells, is the gene for hereditary haemorrhagic telangiectasia type 1","volume":"8","author":"McAllister","year":"1994","journal-title":"Nat. Genet."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"464","DOI":"10.1007\/PL00012413","article-title":"Inducible expression of human endoglin during inflammation and wound healing in vivo","volume":"51","author":"Torsney","year":"2002","journal-title":"Inflamm. Res."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"765","DOI":"10.1042\/bj3010765","article-title":"Phosphorylation of the human-transforming-growth-factor-beta-binding protein endoglin","volume":"301","author":"Lastres","year":"1994","journal-title":"Biochem. J."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"155","DOI":"10.1007\/s10456-016-9495-8","article-title":"Transcription factor KLF6 upregulates expression of metalloprotease MMP14 and subsequent release of soluble endoglin during vascular injury","volume":"19","author":"Blanco","year":"2016","journal-title":"Angiogenesis"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"2483","DOI":"10.1161\/01.STR.0000088644.60368.ED","article-title":"Hypoxic induction of endoglin via mitogen-activated protein kinases in mouse brain microvascular endothelial cells","volume":"34","author":"Zhu","year":"2003","journal-title":"Stroke"},{"key":"ref_15","first-page":"1209","article-title":"MMP-14 aggravates onset of severe preeclampsia by mediating soluble endoglin release","volume":"22","author":"Zhang","year":"2018","journal-title":"Eur. Rev. Med. Pharmacol. Sci."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"4141","DOI":"10.1158\/0008-5472.CAN-09-4466","article-title":"Matrix metalloproteinase-14 (MT1-MMP)-mediated endoglin shedding inhibits tumor angiogenesis","volume":"70","author":"Hawinkels","year":"2010","journal-title":"Cancer Res."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Aristorena, M., Gallardo-Vara, E., Vicen, M., de Las Casas-Engel, M., Ojeda-Fernandez, L., Nieto, C., Blanco, F.J., Valbuena-Diez, A.C., Botella, L.M., and Nachtigal, P. (2019). MMP-12, Secreted by Pro-Inflammatory Macrophages, Targets Endoglin in Human Macrophages and Endothelial Cells. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20123107"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"43799","DOI":"10.1074\/jbc.M207160200","article-title":"Endoglin expression is regulated by transcriptional cooperation between the hypoxia and transforming growth factor-beta pathways","volume":"277","author":"Botella","year":"2002","journal-title":"J. Biol. Chem."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1794","DOI":"10.1161\/01.STR.25.9.1794","article-title":"Role of angiogenesis in patients with cerebral ischemic stroke","volume":"25","author":"Krupinski","year":"1994","journal-title":"Stroke"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1677","DOI":"10.1038\/jcbfm.2012.88","article-title":"Inflammatory cytokines in experimental and human stroke","volume":"32","author":"Lambertsen","year":"2012","journal-title":"J. Cereb. Blood Flow Metab."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1016\/j.cardiores.2005.02.020","article-title":"Cytokine stimulated vascular cell adhesion molecule-1 (VCAM-1) ectodomain release is regulated by TIMP-3","volume":"67","author":"Singh","year":"2005","journal-title":"Cardiovasc. Res."},{"key":"ref_22","first-page":"543","article-title":"E-selectin and intercellular adhesion molecule-1 are released by activated human endothelial cells in vitro","volume":"77","author":"Leeuwenberg","year":"1992","journal-title":"Immunology"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"3970","DOI":"10.1038\/onc.2013.386","article-title":"Antibody-directed coupling of endoglin and MMP-14 is a key mechanism for endoglin shedding and deregulation of TGF-beta signaling","volume":"33","author":"Kumar","year":"2014","journal-title":"Oncogene"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"299","DOI":"10.1002\/jcp.10170","article-title":"Targets of transcriptional regulation by two distinct type I receptors for transforming growth factor-beta in human umbilical vein endothelial cells","volume":"193","author":"Ota","year":"2002","journal-title":"J. Cell Physiol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"2101","DOI":"10.1161\/STROKEAHA.114.005115","article-title":"Endoglin deficiency impairs stroke recovery","volume":"45","author":"Shen","year":"2014","journal-title":"Stroke"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"171","DOI":"10.1042\/CS20060049","article-title":"Can angiogenesis be exploited to improve stroke outcome? Mechanisms and therapeutic potential","volume":"111","author":"Slevin","year":"2006","journal-title":"Clin. Sci."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"642","DOI":"10.1038\/nm1429","article-title":"Soluble endoglin contributes to the pathogenesis of preeclampsia","volume":"12","author":"Venkatesha","year":"2006","journal-title":"Nat. Med."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"52","DOI":"10.1016\/j.lfs.2017.03.014","article-title":"Soluble endoglin modulates the pro-inflammatory mediators NF-kappaB and IL-6 in cultured human endothelial cells","volume":"175","author":"Varejckova","year":"2017","journal-title":"Life Sci."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"323","DOI":"10.1016\/S0021-9150(00)00422-6","article-title":"Endoglin, a TGF-beta receptor-associated protein, is expressed by smooth muscle cells in human atherosclerotic plaques","volume":"153","author":"Conley","year":"2000","journal-title":"Atherosclerosis"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Kim, S.J., Moon, G.J., Cho, Y.H., Kang, H.Y., Hyung, N.K., Kim, D., Lee, J.H., Nam, J.Y., and Bang, O.Y. (2012). Circulating mesenchymal stem cells microparticles in patients with cerebrovascular disease. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0037036"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1296","DOI":"10.1111\/j.1538-7836.2006.01911.x","article-title":"Circulating endothelial microparticles in acute ischemic stroke: A link to severity, lesion volume and outcome","volume":"4","author":"Simak","year":"2006","journal-title":"J. Thromb. Haemost."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"403","DOI":"10.1182\/blood-2012-06-435347","article-title":"Endothelial endoglin is involved in inflammation: Role in leukocyte adhesion and transmigration","volume":"121","author":"Rossi","year":"2013","journal-title":"Blood"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"473","DOI":"10.1038\/s41582-019-0221-1","article-title":"Thrombo-inflammation in acute ischaemic stroke\u2014Implications for treatment","volume":"15","author":"Stoll","year":"2019","journal-title":"Nat. Rev. Neurol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1023","DOI":"10.1161\/CIRCRESAHA.120.316655","article-title":"CD84 Links T Cell and Platelet Activity in Cerebral Thrombo-Inflammation in Acute Stroke","volume":"127","author":"Schuhmann","year":"2020","journal-title":"Circ. Res."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"615","DOI":"10.1001\/jamaneurol.2021.0065","article-title":"Local Cerebral Recombinant Tissue Plasminogen Activator Concentrations During Acute Stroke","volume":"78","author":"Essig","year":"2021","journal-title":"JAMA Neurol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"156","DOI":"10.1186\/s12974-018-1170-0","article-title":"Novel insights into neuroinflammation: Bacterial lipopolysaccharide, tumor necrosis factor alpha, and Ureaplasma species differentially modulate atypical chemokine receptor 3 responses in human brain microvascular endothelial cells","volume":"15","author":"Silwedel","year":"2018","journal-title":"J. Neuroinflamm."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"639","DOI":"10.1007\/s00401-015-1417-0","article-title":"Soluble VCAM-1 impairs human brain endothelial barrier integrity via integrin alpha-4-transduced outside-in signalling","volume":"129","author":"Haarmann","year":"2015","journal-title":"Acta Neuropathol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"23671","DOI":"10.1038\/srep23671","article-title":"Pitfalls in assessing microvascular endothelial barrier function: Impedance-based devices versus the classic macromolecular tracer assay","volume":"6","author":"Bischoff","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Kilkenny, C., Browne, W.J., Cuthill, I.C., Emerson, M., and Altman, D.G. (2010). Improving bioscience research reporting: The ARRIVE guidelines for reporting animal research. PLoS Biol., 8.","DOI":"10.1371\/journal.pbio.1000412"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"46","DOI":"10.1186\/s12974-021-02095-1","article-title":"Platelets and lymphocytes drive progressive penumbral tissue loss during middle cerebral artery occlusion in mice","volume":"18","author":"Schuhmann","year":"2021","journal-title":"J. Neuroinflamm."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/13\/7085\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T23:38:30Z","timestamp":1760139510000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/13\/7085"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,6,25]]},"references-count":40,"journal-issue":{"issue":"13","published-online":{"date-parts":[[2022,7]]}},"alternative-id":["ijms23137085"],"URL":"https:\/\/doi.org\/10.3390\/ijms23137085","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,6,25]]}}}