{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T15:33:04Z","timestamp":1788449584290,"version":"build-2803163510"},"reference-count":45,"publisher":"MDPI AG","issue":"22","license":[{"start":{"date-parts":[[2025,11,12]],"date-time":"2025-11-12T00:00:00Z","timestamp":1762905600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"crossref","award":["NS-106746"],"award-info":[{"award-number":["NS-106746"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"crossref","award":["NS-116786"],"award-info":[{"award-number":["NS-116786"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Subarachnoid hemorrhage (SAH) is a devastating cerebrovascular disorder with high mortality and long-term disability. It is more prevalent in women than men, but most preclinical research has been performed in male animals. Upregulation of lipocalin-2 (Lcn2), an acute-phase protein involved in iron homeostasis and neuroinflammation, has been implicated in hemorrhagic brain injury in male animals. The purpose of this study was to examine whether genetic deletion of Lcn2 also reduces early brain injury after SAH in female mice. Adult female wild-type (WT) and Lcn2 knockout (KO) mice were subjected to endovascular perforation to induce SAH. Lcn2 expression was assessed by immunohistochemistry and Western blotting, while brain injury was evaluated using MRI T2 lesion measurement, blood\u2013brain barrier (BBB) permeability assays, Fluoro-Jade C staining, and Garcia\u2019s neurological scoring. We found that Lcn2 expression was upregulated in multiple brain regions after SAH, particularly in astrocytes. Compared with WT mice, Lcn2 KO mice exhibited significantly reduced oxidative stress, attenuated ferritin induction, smaller T2 lesions, decreased BBB leakage, reduced neuronal degeneration, and improved neurological recovery over 7 days. These findings identify Lcn2 as a critical mediator of early brain injury after SAH in female mice. These results further support targeting Lcn2 as a therapeutic strategy to reduce brain damage and improve outcomes in SAH patients.<\/jats:p>","DOI":"10.3390\/cells14221770","type":"journal-article","created":{"date-parts":[[2025,11,13]],"date-time":"2025-11-13T09:10:45Z","timestamp":1763025045000},"page":"1770","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Role of Lipocalin-2 in Brain Injury After Subarachnoid Hemorrhage in Female Mice"],"prefix":"10.3390","volume":"14","author":[{"given":"Hao","family":"Zhao","sequence":"first","affiliation":[{"name":"Department of Neurosurgery, University of Michigan, Ann Arbor, MI 48109, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1176-2065","authenticated-orcid":false,"given":"Yingfeng","family":"Wan","sequence":"additional","affiliation":[{"name":"Department of Neurosurgery, University of Michigan, Ann Arbor, MI 48109, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sravanthi","family":"Koduri","sequence":"additional","affiliation":[{"name":"Department of Neurosurgery, University of Michigan, Ann Arbor, MI 48109, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0100-2289","authenticated-orcid":false,"given":"Ya","family":"Hua","sequence":"additional","affiliation":[{"name":"Department of Neurosurgery, University of Michigan, Ann Arbor, MI 48109, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guohua","family":"Xi","sequence":"additional","affiliation":[{"name":"Department of Neurosurgery, University of Michigan, Ann Arbor, MI 48109, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6441-5334","authenticated-orcid":false,"given":"Richard F.","family":"Keep","sequence":"additional","affiliation":[{"name":"Department of Neurosurgery, University of Michigan, Ann Arbor, MI 48109, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,11,12]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"846","DOI":"10.1016\/S0140-6736(22)00938-2","article-title":"Spontaneous subarachnoid haemorrhage","volume":"400","author":"Claassen","year":"2022","journal-title":"Lancet"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"588","DOI":"10.1001\/jamaneurol.2019.0006","article-title":"Worldwide Incidence of Aneurysmal Subarachnoid Hemorrhage According to Region, Time Period, Blood Pressure, and Smoking Prevalence in the Population: A Systematic Review and Meta-analysis","volume":"76","author":"Etminan","year":"2019","journal-title":"JAMA Neurol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1347","DOI":"10.1161\/STROKEAHA.122.041488","article-title":"Sex and Racial Disparity in Outcome of Aneurysmal Subarachnoid Hemorrhage in the United States: A 20-Year Analysis","volume":"54","author":"Schupper","year":"2023","journal-title":"Stroke"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"210","DOI":"10.1124\/pr.107.08002","article-title":"Vascular actions of estrogens: Functional implications","volume":"60","author":"Miller","year":"2008","journal-title":"Pharmacol. Rev."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"558","DOI":"10.1177\/1099800414561632","article-title":"Plasma Estrogen Levels Are Associated With Severity of Injury and Outcomes After Aneurysmal Subarachnoid Hemorrhage","volume":"17","author":"Crago","year":"2015","journal-title":"Biol. Res. Nurs."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1426","DOI":"10.1161\/STROKEAHA.122.040072","article-title":"Early Brain Injury After Subarachnoid Hemorrhage: Incidence and Mechanisms","volume":"54","author":"Lauzier","year":"2023","journal-title":"Stroke"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"109086","DOI":"10.1016\/j.clineuro.2025.109086","article-title":"Dynamic correlation between brain edema scores and blood\u2013brain barrier permeability predicts the outcome of aneurysmal subarachnoid hemorrhage","volume":"257","author":"Zhang","year":"2025","journal-title":"Clin. Neurol. Neurosurg."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1007\/s11910-024-01396-1","article-title":"Update on Strategies to Reduce Early Brain Injury after Subarachnoid Hemorrhage","volume":"25","author":"Yang","year":"2024","journal-title":"Curr. Neurol. Neurosci. Rep."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Galaris, A., Fanidis, D., Tsitoura, E., Kanellopoulou, P., Barbayianni, I., Ntatsoulis, K., Touloumi, K., Gramenoudi, S., Karampitsakos, T., and Tzouvelekis, A. (2023). Increased lipocalin-2 expression in pulmonary inflammation and fibrosis. Front. Med., 10.","DOI":"10.3389\/fmed.2023.1195501"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"669","DOI":"10.5483\/BMBRep.2023-0177","article-title":"SARS-CoV-2 infection induces expression and secretion of lipocalin-2 and regulates iron in a human lung cancer xenograft model","volume":"56","author":"Park","year":"2023","journal-title":"BMB Rep."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Zhao, R.Y., Wei, P.J., Sun, X., Zhang, D.H., He, Q.Y., Liu, J., Chang, J.L., Yang, Y., and Guo, Z.N. (2023). Role of lipocalin 2 in stroke. Neurobiol. Dis., 179.","DOI":"10.1016\/j.nbd.2023.106044"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"103","DOI":"10.1146\/annurev-nutr-071816-064559","article-title":"Lipocalin 2: An Emerging Player in Iron Homeostasis and Inflammation","volume":"37","author":"Xiao","year":"2017","journal-title":"Annu. Rev. Nutr."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1454","DOI":"10.1038\/jcbfm.2015.52","article-title":"Role of lipocalin-2 in brain injury after intracerebral hemorrhage","volume":"35","author":"Ni","year":"2015","journal-title":"J. Cereb. Blood Flow Metab."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1078","DOI":"10.1161\/STROKEAHA.115.012153","article-title":"Role of Lipocalin-2 in Thrombin-Induced Brain Injury","volume":"47","author":"Mao","year":"2016","journal-title":"Stroke"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"2524","DOI":"10.1177\/0271678X211012110","article-title":"CSF lipocalin-2 increases early in subarachnoid hemorrhage are associated with neuroinflammation and unfavorable outcome","volume":"41","author":"Yu","year":"2021","journal-title":"J. Cereb. Blood Flow Metab."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1016\/j.jneumeth.2015.01.035","article-title":"A magnetic resonance imaging grading system for subarachnoid hemorrhage severity in a rat model","volume":"243","author":"Shishido","year":"2015","journal-title":"J. Neurosci. Methods"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"582","DOI":"10.1161\/STROKEAHA.114.007834","article-title":"New grading system based on magnetic resonance imaging in a mouse model of subarachnoid hemorrhage","volume":"46","author":"Egashira","year":"2015","journal-title":"Stroke"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"627","DOI":"10.1161\/01.STR.26.4.627","article-title":"Neurological deficit and extent of neuronal necrosis attributable to middle cerebral artery occlusion in rats. Statistical validation","volume":"26","author":"Garcia","year":"1995","journal-title":"Stroke"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2141","DOI":"10.1161\/STROKEAHA.114.005307","article-title":"Acute white matter injury after experimental subarachnoid hemorrhage: Potential role of lipocalin 2","volume":"45","author":"Egashira","year":"2014","journal-title":"Stroke"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1207","DOI":"10.1111\/cns.13221","article-title":"White matter T2 hyperintensities and blood-brain barrier disruption in the hyperacute stage of subarachnoid hemorrhage in male mice: The role of lipocalin-2","volume":"25","author":"Toyota","year":"2019","journal-title":"CNS Neurosci. Ther."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Anzabi, M., Ardalan, M., Iversen, N.K., Rafati, A.H., Hansen, B., and Ostergaard, L. (2018). Hippocampal Atrophy Following Subarachnoid Hemorrhage Correlates with Disruption of Astrocyte Morphology and Capillary Coverage by AQP4. Front. Cell. Neurosci., 12.","DOI":"10.3389\/fncel.2018.00019"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Regnier-Golanov, A.S., Dundar, F., Zumbo, P., Betel, D., Hernandez, M.S., Peterson, L.E., Lo, E.H., Golanov, E.V., and Britz, G.W. (2021). Hippocampal Transcriptome Changes After Subarachnoid Hemorrhage in Mice. Front. Neurol., 12.","DOI":"10.3389\/fneur.2021.691631"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"43855","DOI":"10.1074\/jbc.M111.299248","article-title":"Lipocalin-2 Is a chemokine inducer in the central nervous system: Role of chemokine ligand 10 (CXCL10) in lipocalin-2-induced cell migration","volume":"286","author":"Lee","year":"2011","journal-title":"J. Biol. Chem."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"330","DOI":"10.1186\/s12974-018-1372-5","article-title":"Lipocalin 2 contributes to brain iron dysregulation but does not affect cognition, plaque load, and glial activation in the J20 Alzheimer mouse model","volume":"15","author":"Dekens","year":"2018","journal-title":"J. Neuroinflamm."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"168","DOI":"10.1186\/s12974-019-1556-7","article-title":"Lipocalin-2 may produce damaging effect after cerebral ischemia by inducing astrocytes classical activation","volume":"16","author":"Zhao","year":"2019","journal-title":"J. Neuroinflamm."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"86","DOI":"10.1016\/j.brainres.2013.02.008","article-title":"Role of iron in brain lipocalin 2 upregulation after intracerebral hemorrhage in rats","volume":"1505","author":"Dong","year":"2013","journal-title":"Brain Res."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"546","DOI":"10.1007\/s12975-013-0270-5","article-title":"Deferoxamine reduces neuronal death and hematoma lysis after intracerebral hemorrhage in aged rats","volume":"4","author":"Hatakeyama","year":"2013","journal-title":"Transl. Stroke Res."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"130","DOI":"10.1007\/s12975-012-0232-3","article-title":"DARPP-32 to quantify intracerebral hemorrhage-induced neuronal death in basal ganglia","volume":"4","author":"Jin","year":"2013","journal-title":"Transl. Stroke Res."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1615\/CritRevImmunol.2015012127","article-title":"Lipocalin-2 in the Inflammatory Activation of Brain Astrocytes","volume":"35","author":"Lee","year":"2015","journal-title":"Crit. Rev. Immunol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"591","DOI":"10.1016\/j.neuint.2011.04.007","article-title":"Expression and localization of the iron-siderophore binding protein lipocalin 2 in the normal rat brain and after kainate-induced excitotoxicity","volume":"59","author":"Chia","year":"2011","journal-title":"Neurochem. Int."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"13412","DOI":"10.1523\/JNEUROSCI.0116-11.2011","article-title":"Lipocalin 2 plays an immunomodulatory role and has detrimental effects after spinal cord injury","volume":"31","author":"Rathore","year":"2011","journal-title":"J. Neurosci."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1293","DOI":"10.1016\/j.cell.2005.10.027","article-title":"A cell-surface receptor for lipocalin 24p3 selectively mediates apoptosis and iron uptake","volume":"123","author":"Devireddy","year":"2005","journal-title":"Cell"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"234","DOI":"10.1523\/JNEUROSCI.5273-08.2009","article-title":"Lipocalin-2 is an autocrine mediator of reactive astrocytosis","volume":"29","author":"Lee","year":"2009","journal-title":"J. Neurosci."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Shin, H.J., Jeong, E.A., Lee, J.Y., An, H.S., Jang, H.M., Ahn, Y.J., Lee, J., Kim, K.E., and Roh, G.S. (2021). Lipocalin-2 Deficiency Reduces Oxidative Stress and Neuroinflammation and Results in Attenuation of Kainic Acid-Induced Hippocampal Cell Death. Antioxidants, 10.","DOI":"10.3390\/antiox10010100"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1183","DOI":"10.1210\/en.2011-1642","article-title":"Lipocalin 2 deficiency alters estradiol production and estrogen receptor signaling in female mice","volume":"153","author":"Guo","year":"2012","journal-title":"Endocrinology"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"5566","DOI":"10.1074\/jbc.M114.606459","article-title":"Estrogen receptor (ER)alpha-regulated lipocalin 2 expression in adipose tissue links obesity with breast cancer progression","volume":"290","author":"Drew","year":"2015","journal-title":"J. Biol. Chem."},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Krizanac, M., Mass Sanchez, P.B., Weiskirchen, R., and Schroder, S.K. (2024). Overview of the expression patterns and roles of Lipocalin 2 in the reproductive system. Front. Endocrinol., 15.","DOI":"10.3389\/fendo.2024.1365602"},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Fiocchetti, M., Ascenzi, P., and Marino, M. (2012). Neuroprotective effects of 17beta-estradiol rely on estrogen receptor membrane initiated signals. Front. Physiol., 3.","DOI":"10.3389\/fphys.2012.00073"},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Stragier, H., Vandersmissen, H., Ordies, S., Thiessen, S., Mesotten, D., Peuskens, D., and Ten Cate, H. (2025). Pathophysiological mechanisms underlying early brain injury and delayed cerebral ischemia in the aftermath of aneurysmal subarachnoid hemorrhage: A comprehensive analysis. Front. Neurol., 16.","DOI":"10.3389\/fneur.2025.1587091"},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"de Oliveira, J., Denadai, M.B., and Costa, D.L. (2022). Crosstalk between Heme Oxygenase-1 and Iron Metabolism in Macrophages: Implications for the Modulation of Inflammation and Immunity. Antioxidants, 11.","DOI":"10.3390\/antiox11050861"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"337","DOI":"10.1097\/SHK.0000000000000107","article-title":"Regulation of iron uptake in primary culture rat hepatocytes: The role of acute-phase cytokines","volume":"41","author":"Ahmad","year":"2014","journal-title":"Shock"},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Xiao, R., Pan, J., Yang, M., Liu, H., Zhang, A., Guo, X., and Zhou, S. (2025). Regulating astrocyte phenotype by Lcn2 inhibition toward ischemic stroke therapy. Biomaterials, 317.","DOI":"10.1016\/j.biomaterials.2025.123102"},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Zheng, N., Li, X., Zhou, N., and Luo, L. (2025). Identification of Novel LCN2 Inhibitors Based on Construction of Pharmacophore Models and Screening of Marine Compound Libraries by Fragment Design. Mar. Drugs, 23.","DOI":"10.3390\/md23010024"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"158","DOI":"10.1016\/j.pneurobio.2016.08.001","article-title":"Lipocalin-2 as a therapeutic target for brain injury: An astrocentric perspective","volume":"144","author":"Suk","year":"2016","journal-title":"Prog. Neurobiol."},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"Wang, G., Weng, Y.C., Chiang, I.C., Huang, Y.T., Liao, Y.C., Chen, Y.C., Kao, C.Y., Liu, Y.L., Lee, T.H., and Chou, W.H. (2020). Neutralization of Lipocalin-2 Diminishes Stroke-Reperfusion Injury. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21176253"}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/14\/22\/1770\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,11,14]],"date-time":"2025-11-14T05:30:36Z","timestamp":1763098236000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/14\/22\/1770"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,11,12]]},"references-count":45,"journal-issue":{"issue":"22","published-online":{"date-parts":[[2025,11]]}},"alternative-id":["cells14221770"],"URL":"https:\/\/doi.org\/10.3390\/cells14221770","relation":{},"ISSN":["2073-4409"],"issn-type":[{"value":"2073-4409","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,11,12]]}}}