{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T00:43:22Z","timestamp":1759970602931,"version":"build-2065373602"},"reference-count":47,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2025,1,26]],"date-time":"2025-01-26T00:00:00Z","timestamp":1737849600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Funda\u00e7\u00e3o Carlos Chagas Filho de Amparo \u00e0 Pesquisa do Estado do Rio de Janeiro (FAPERJ)"},{"name":"Coordena\u00e7\u00e3o de Aperfei\u00e7oamento de Pessoal de N\u00edvel Superior (CAPES)"},{"name":"Conselho Nacional de Desenvolvimento Cient\u00edfico e Tecnol\u00f3gico (CNPq)"},{"name":"National Institute of Science and Technology on Neuroimmunomodulation (INCT-NIM)"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Brain Sciences"],"abstract":"<jats:p>Background: Ouabain is a steroid hormone that binds to the sodium pump (Na+, K+-ATPase) at physiological (nanomolar) concentrations, activating different signaling pathways. This interaction has been shown to prevent the axotomy-induced death of retinal ganglion cells (RGCs), although the underlying mechanisms remain unclear. Objective: In this study, we investigated potential mechanisms by which ouabain promotes RGC survival using primary cultures of rat neural retina. Results: Our findings indicate that ouabain regulates brain-derived neurotrophic factor (BDNF) signaling in retinal cells via matrix metalloproteinase-9-mediated processing of proBDNF to mature BDNF (mBDNF) and by increasing the phosphorylation of the mBDNF receptor, tropomyosin-related receptor kinase B. Ouabain also enhances the maturation of interleukin (IL)-1\u03b2 through the increased activation of caspase-1, which mediates the processing of proIL-1\u03b2 into IL-1\u03b2, and transiently upregulates both IL-1 receptor and IL-1 receptor antagonist (IL-1Ra). Treatment using either IL-1\u03b2 or IL-1Ra alone is sufficient to enhance RGC survival similarly to that achieved with ouabain. Finally, we further show that ouabain prevents RGC death through a complex signaling mechanism shared by BDNF and IL-1\u03b2, which includes the activation of the Src and protein kinase C pathways. Conclusions: Collectively, these results suggest that ouabain stimulates the maturation and signaling of both BDNF and IL-1\u03b2, which act as key mediators of RGC survival.<\/jats:p>","DOI":"10.3390\/brainsci15020123","type":"journal-article","created":{"date-parts":[[2025,1,27]],"date-time":"2025-01-27T03:54:27Z","timestamp":1737950067000},"page":"123","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Ouabain Counteracts Retinal Ganglion Cell Death Through Modulation of BDNF and IL-1 Signaling Pathways"],"prefix":"10.3390","volume":"15","author":[{"given":"Amanda Candida da Rocha","family":"Oliveira","sequence":"first","affiliation":[{"name":"Department of Neurobiology and Program of Neurosciences, Institute of Biology, Federal Fluminense University, Niteroi 24020-141, Brazil"},{"name":"National Institute of Science and Technology on Neuroimmunomodulation\u2014INCT-NIM, Oswaldo Cruz Foundation, Rio de Janeiro 21041-250, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5262-231X","authenticated-orcid":false,"given":"Camila Saggioro","family":"Figueiredo","sequence":"additional","affiliation":[{"name":"Department of Neurobiology and Program of Neurosciences, Institute of Biology, Federal Fluminense University, Niteroi 24020-141, Brazil"},{"name":"National Institute of Science and Technology on Neuroimmunomodulation\u2014INCT-NIM, Oswaldo Cruz Foundation, Rio de Janeiro 21041-250, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7037-5794","authenticated-orcid":false,"given":"\u00cdcaro","family":"Raony","sequence":"additional","affiliation":[{"name":"Institute of Medical Biochemistry Leopoldo de Meis, Federal University of Rio de Janeiro, Rio de Janeiro 21941-902, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Juliana Salles","family":"Von-Held-Ventura","sequence":"additional","affiliation":[{"name":"Department of Neurobiology and Program of Neurosciences, Institute of Biology, Federal Fluminense University, Niteroi 24020-141, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6243-0768","authenticated-orcid":false,"given":"Marcelo Gomes","family":"Granja","sequence":"additional","affiliation":[{"name":"Research, Innovation, and Surveillance Center for COVID-19 and Health Emergencies, Oswaldo Cruz Foundation, Rio de Janeiro 21040-360, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Thalita","family":"M\u00e1zala-de-Oliveira","sequence":"additional","affiliation":[{"name":"Department of Neurobiology and Program of Neurosciences, Institute of Biology, Federal Fluminense University, Niteroi 24020-141, Brazil"},{"name":"National Institute of Science and Technology on Neuroimmunomodulation\u2014INCT-NIM, Oswaldo Cruz Foundation, Rio de Janeiro 21041-250, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Vin\u00edcius Henrique","family":"Pedrosa-Soares","sequence":"additional","affiliation":[{"name":"Department of Neurobiology and Program of Neurosciences, Institute of Biology, Federal Fluminense University, Niteroi 24020-141, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4199-9821","authenticated-orcid":false,"given":"Aline Araujo","family":"dos Santos","sequence":"additional","affiliation":[{"name":"National Institute of Science and Technology on Neuroimmunomodulation\u2014INCT-NIM, Oswaldo Cruz Foundation, Rio de Janeiro 21041-250, Brazil"},{"name":"Department of Physiology and Pharmacology, Biomedical Institute, Federal Fluminense University, Niteroi 24210-130, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Elizabeth","family":"Giestal-de-Araujo","sequence":"additional","affiliation":[{"name":"Department of Neurobiology and Program of Neurosciences, Institute of Biology, Federal Fluminense University, Niteroi 24020-141, Brazil"},{"name":"National Institute of Science and Technology on Neuroimmunomodulation\u2014INCT-NIM, Oswaldo Cruz Foundation, Rio de Janeiro 21041-250, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,1,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"218","DOI":"10.1038\/eye.2016.299","article-title":"Protecting retinal ganglion cells","volume":"31","author":"Khatib","year":"2017","journal-title":"Eye"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Tempone, M.H., Borges-Martins, V.P., C\u00e9sar, F., Alexandrino-Mattos, D.P., de Figueiredo, C.S., Raony, \u00cd., dos Santos, A.A., Duarte-Silva, A.T., Dias, M.S., and Freitas, H.R. (2024). The healthy and diseased retina seen through neuron\u2013glia interactions. Int. J. Mol. Sci., 25.","DOI":"10.20944\/preprints202312.1868.v2"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"187","DOI":"10.1007\/978-1-4939-7407-8_16","article-title":"In vitro and in vivo methods for studying retinal ganglion cell survival and optic nerve regeneration","volume":"1695","author":"Yin","year":"2018","journal-title":"Methods Mol. Biol."},{"key":"ref_4","unstructured":"Vision Loss Expert Group of the Global Burden of Disease Study (2024). Global estimates on the number of people blind or visually impaired by glaucoma: A meta-analysis from 2000 to 2020. Eye, 38, 2036\u20132046."},{"key":"ref_5","first-page":"89","article-title":"Ouabain induces an increase of retinal ganglion cell survival in vitro: The involvement of protein kinase C","volume":"5","author":"Fontes","year":"2005","journal-title":"Brain Res."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1343","DOI":"10.1007\/s11064-010-0190-7","article-title":"The trophic effect of ouabain on retinal ganglion cell is mediated by EGF receptor and PKC \u03b4 activation","volume":"35","year":"2010","journal-title":"Neurochem. Res."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"651","DOI":"10.1007\/s10571-015-0160-3","article-title":"Ouabain and BDNF crosstalk on ganglion cell survival in mixed retinal cell cultures","volume":"35","author":"Soares","year":"2015","journal-title":"Cell Mol. Neurobiol."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Lambuk, L., Mohd Lazaldin, M.A., Ahmad, S., Iezhitsa, I., Agarwal, R., Uskokovi\u0107, V., and Mohamud, R. (2022). Brain-derived neurotrophic factor mediated neuroprotection in glaucoma: A review of current state of the art. Front. Pharmacol., 20.","DOI":"10.3389\/fphar.2022.875662"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"17023","DOI":"10.1038\/sigtrans.2017.23","article-title":"NF-\u03baB signaling in inflammation","volume":"2","author":"Liu","year":"2017","journal-title":"Sig. Transduct Target Ther."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1092","DOI":"10.1016\/j.stemcr.2024.06.009","article-title":"Human microglia-derived proinflammatory cytokines facilitate human retinal ganglion cell development and regeneration","volume":"19","author":"Subramani","year":"2024","journal-title":"Stem Cell Rep."},{"key":"ref_11","first-page":"378","article-title":"The trophic effect of ouabain on retinal ganglion cells is mediated by IL-1\u03b2 and TNF-\u03b1","volume":"478","author":"Granja","year":"2016","journal-title":"BBRC"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"723","DOI":"10.1007\/s11064-021-03481-0","article-title":"Ouabain-Na+\/K+-ATPase signaling regulates retinal neuroinflammation and ROS production preventing neuronal death by an autophagy-dependent mechanism following optic nerve axotomy in vitro","volume":"47","author":"Miranda","year":"2022","journal-title":"Neurochem. Res."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1701","DOI":"10.1007\/s11064-021-03313-1","article-title":"Increased retinal ganglion cell survival by exogenous IL-2 depends on IL-10, dopamine D1 receptors, and classical IL-2\/IL-2R signaling pathways","volume":"46","author":"Colares","year":"2021","journal-title":"Neurochem. Res."},{"key":"ref_14","unstructured":"Mesulam, M. (1982). Tracing Neural Connections with Horseradish Peroxidase, Wiley. [1st ed.]."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"549","DOI":"10.1002\/j.1460-2075.1982.tb01207.x","article-title":"Purification of a new neurotrophic factor from mammalian brain","volume":"1","author":"Barde","year":"1982","journal-title":"EMBO J."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"299","DOI":"10.1038\/nrn1078","article-title":"Neurotrophins and their receptors: A convergence point for many signalling pathways","volume":"4","author":"Chao","year":"2003","journal-title":"Nat. Rev. Neurosci."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"321","DOI":"10.1002\/jez.1401160206","article-title":"Selective growth stimulating effects of mouse sarcoma on the sensory and sympathetic nervous system of the chick embryo","volume":"116","author":"Hamburger","year":"1951","journal-title":"J. Exp. Zool."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"603","DOI":"10.1038\/nrn1726","article-title":"The yin and yang of neurotrophin action","volume":"6","author":"Lu","year":"2005","journal-title":"Nat. Rev. Neurosci."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Reinhard, S.M., Razak, K., and Ethell, I.M. (2015). A delicate balance: Role of MMP-9 in brain development and pathophysiology of neurodevelopmental disorders. Front. Cell Neurosci., 29.","DOI":"10.3389\/fncel.2015.00280"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"11995","DOI":"10.1074\/jbc.M403172200","article-title":"Activation of the Trk signaling pathway by extracellular zinc: Role of metalloproteinases","volume":"280","author":"Hwang","year":"2005","journal-title":"JBC"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"4643","DOI":"10.1038\/s41467-021-24810-5","article-title":"Stress-primed secretory autophagy promotes extracellular BDNF maturation by enhancing MMP9 secretion","volume":"12","author":"Martinelli","year":"2021","journal-title":"Nat. Commun."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"232","DOI":"10.1038\/nrrheum.2010.4","article-title":"IL-1 pathways in inflammation and human diseases","volume":"6","author":"Gabay","year":"2010","journal-title":"Nat. Rev. Rheumatol."},{"key":"ref_23","first-page":"489","article-title":"Effects of axotomy and intraocular administration of NT-4, NT-3 and Brain-derived neurotrophic factor on the survival of adult rat retinal ganglion cells: A quantitative In Vivo Study","volume":"37","author":"Salvador","year":"1996","journal-title":"Invest. Ophthalmol. Vis. Sci."},{"key":"ref_24","first-page":"135","article-title":"Effects of neurotrophins on the survival and regrowth of injured retinal neurons","volume":"196","author":"Aguayo","year":"1996","journal-title":"Ciba Found. Symp."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1038","DOI":"10.1523\/JNEUROSCI.18-03-01038.1998","article-title":"Free radical scavenging and inhibition of nitric oxide synthase potentiates the neurotrophic effect of brain-derived neurotrophic factor on axotomized retinal ganglion cells in vivo","volume":"18","author":"Cellerino","year":"1998","journal-title":"J. Neurosci."},{"key":"ref_26","first-page":"966","article-title":"BDNF enhances retinal ganglion cell survival in cats with optic nerve damage","volume":"42","author":"Chen","year":"2001","journal-title":"Invest. Ophthalmol. Vis. Sci."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1632","DOI":"10.1073\/pnas.91.5.1632","article-title":"Effects of ocular injury and administration of Brain-derived neurotrophic factor on survival and regrowth of axotomized retinal ganglion cells","volume":"91","author":"Clarke","year":"1994","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"304","DOI":"10.1016\/0006-8993(93)90695-J","article-title":"Intravitreal injections of neurotrophic factors support the survival of axotomized retinal ganglion cells in adult rats in vivo","volume":"602","author":"Mey","year":"1993","journal-title":"Brain Res."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"32","DOI":"10.1016\/j.exer.2009.02.015","article-title":"Effects of different neurotrophic factors on the survival of retinal ganglion cells after a complete intraorbital nerve crush injury: A quantitative in vivo study","volume":"89","author":"Agudo","year":"2009","journal-title":"Exp. Eye Res."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Kimura, A., Namekata, K., Guo, X., Harada, C., and Harada, T. (2016). Neuroprotection, growth factors, and BDNF-TrkB signalling in retinal degeneration. Int. J. Mol. Sci., 17.","DOI":"10.3390\/ijms17091584"},{"key":"ref_31","first-page":"3460","article-title":"Retrograde axonal transport of BDNF in retinal ganglion cells is blocked by acute IOP elevation in rats","volume":"41","author":"Quigley","year":"2000","journal-title":"Invest. Ophthalmol. Vis. Sci."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Oddone, F., Roberti, G., Micera, A., Busanello, A., Bonini, S., and Quaranta, L. (2017). Exploring serum levels of Brain-derived neurotrophic factor and nerve growth factor across glaucoma stages. PLoS ONE, 12.","DOI":"10.1371\/journal.pone.0168565"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1173","DOI":"10.1007\/s00417-018-3923-y","article-title":"The effect of trabeculectomy on serum Brain-derived neurotrophic factor levels in primary open-angle glaucoma","volume":"256","author":"Uzel","year":"2018","journal-title":"Graefes Arch. Clin. Exp. Ophthalmol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"96","DOI":"10.1016\/0304-3940(94)11123-Z","article-title":"Expression of Brain-derived neurotrophic factor and its functional receptor in neonatal and adult rat retina","volume":"183","author":"Caminos","year":"1995","journal-title":"Neurosci. Lett."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"39392","DOI":"10.1074\/jbc.M110.147801","article-title":"Chronic and acute models of retinal neurodegeneration TrkA activity are neuroprotective whereas p75NTR activity is neurotoxic through a paracrine mechanism","volume":"285","author":"Bai","year":"2010","journal-title":"JBC"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"3977","DOI":"10.1523\/JNEUROSCI.22-10-03977.2002","article-title":"TrkB gene transfer protects retinal ganglion cells from axotomy-induced death in vivo","volume":"22","author":"Cheng","year":"2002","journal-title":"J. Neurosci."},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Orellana, A.M., Kinoshita, P.F., Leite, J.A., Kawamoto, E.M., and Scavone, C. (2016). Cardiotonic steroids as modulators of neuroinfammation. Front. Endocrinol., 7.","DOI":"10.3389\/fendo.2016.00010"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"34","DOI":"10.1016\/j.brainres.2013.09.052","article-title":"Anti-inflammatory substances can influence some glial cell types but not others","volume":"1539","author":"Forshammar","year":"2013","journal-title":"Brain Res."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"218","DOI":"10.1186\/s12974-014-0218-z","article-title":"Signaling function of Na, K-ATPase induced by ouabain against LPS as an inflammation model in hippocampus","volume":"11","author":"Kinoshita","year":"2014","journal-title":"J. Neuroinflammation"},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Wooff, Y., Man, S.M., Aggio-Bruce, R., Natoli, R., and Fernando, N. (2019). IL-1 Family members mediate cell death, inflammation and angiogenesis in retinal degenerative diseases. Front. Immunol., 10.","DOI":"10.3389\/fimmu.2019.01618"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"118","DOI":"10.1186\/s12974-019-1505-5","article-title":"Reactive microglia and IL1\u03b2\/IL-1R1-signaling mediate neuroprotection in excitotoxin-damaged mouse retina","volume":"16","author":"Todd","year":"2019","journal-title":"J. Neuroinflammation"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"e21735","DOI":"10.7554\/eLife.21735","article-title":"Lack of IL-1R8 in neurons causes hyperactivation of IL-1 receptor pathway and induces MECP2-dependent synaptic defects","volume":"6","author":"Tomasoni","year":"2017","journal-title":"eLife"},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Ma, X. (2016). The interleukin-1 family. Regulation of Cytokine Gene Expression in Immunity and Diseases. Advances in Experimental Medicine and Biology, Springer.","DOI":"10.1007\/978-94-024-0921-5"},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Coughlin, B.A., Christian, B., Trombley, B., and Mohr, S. (2024). Interleukin-1 receptor-dependent and-independent caspase-1 activity in retinal cells mediated by receptor interacting protein 2. Front. Cell Dev. Biol., 12.","DOI":"10.3389\/fcell.2024.1467799"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"136197","DOI":"10.1016\/j.neulet.2021.136197","article-title":"PMA treatment fosters rat retinal ganglion cell survival via TNF signaling","volume":"763","author":"Ferreira","year":"2021","journal-title":"Neurosci. Lett."},{"key":"ref_46","first-page":"917","article-title":"PKC delta activation increases neonatal rat retinal cells survival in vitro: Involvement of neurotrophins and M1 muscarinic receptors","volume":"500","author":"Braga","year":"2018","journal-title":"BBRC"},{"key":"ref_47","doi-asserted-by":"crossref","unstructured":"Saba-El-Leil, M.K., Fr\u00e9min, C., and Meloche, S. (2016). Redundancy in the world of MAP kinases: All for one. Front. Cell Dev. Biol., 4.","DOI":"10.3389\/fcell.2016.00067"}],"container-title":["Brain Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-3425\/15\/2\/123\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,8]],"date-time":"2025-10-08T10:36:36Z","timestamp":1759919796000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-3425\/15\/2\/123"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,1,26]]},"references-count":47,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2025,2]]}},"alternative-id":["brainsci15020123"],"URL":"https:\/\/doi.org\/10.3390\/brainsci15020123","relation":{},"ISSN":["2076-3425"],"issn-type":[{"type":"electronic","value":"2076-3425"}],"subject":[],"published":{"date-parts":[[2025,1,26]]}}}