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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

Ouabain is a classic Na + K + ATPase ligand and it has been described to have neuroprotective effects on neurons and glial cells at nanomolar concentrations. In the present work, the neuroprotective and immunomodulatory potential of ouabain was evaluated in neonatal rat retinal cells using an optic...

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Published in:Neurochemical research 2022-03, Vol.47 (3), p.723-738
Main Authors: Mázala-de-Oliveira, Thalita, de Figueiredo, Camila Saggioro, de Rezende Corrêa, Gustavo, da Silva, Mayra Santos, Miranda, Renan Lyra, de Azevedo, Mariana Almeida, Cossenza, Marcelo, dos Santos, Aline Araujo, Giestal-de-Araujo, Elizabeth
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cited_by cdi_FETCH-LOGICAL-c424t-88120f15338be5c901767ea85164dc2f55cac96ab543c37a9aa80a249cc02d663
cites cdi_FETCH-LOGICAL-c424t-88120f15338be5c901767ea85164dc2f55cac96ab543c37a9aa80a249cc02d663
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container_title Neurochemical research
container_volume 47
creator Mázala-de-Oliveira, Thalita
de Figueiredo, Camila Saggioro
de Rezende Corrêa, Gustavo
da Silva, Mayra Santos
Miranda, Renan Lyra
de Azevedo, Mariana Almeida
Cossenza, Marcelo
dos Santos, Aline Araujo
Giestal-de-Araujo, Elizabeth
description Ouabain is a classic Na + K + ATPase ligand and it has been described to have neuroprotective effects on neurons and glial cells at nanomolar concentrations. In the present work, the neuroprotective and immunomodulatory potential of ouabain was evaluated in neonatal rat retinal cells using an optic nerve axotomy model in vitro. After axotomy, cultured retinal cells were treated with ouabain (3 nM) at different periods. The levels of important inflammatory receptors in the retina such as TNFR1/2, TLR4, and CD14 were analyzed. We observed that TNFR1, TLR4, and CD14 were decreased in all tested periods (15 min, 45 min, 24 h, and 48 h). On the other hand, TNFR2 was increased after 24 h, suggesting an anti-inflammatory potential for ouabain. Moreover, we showed that ouabain also decreased Iba-1 (microglial marker) density. Subsequently, analyses of retrograde labeling of retinal ganglion cells (RGC) were performed after 48 h and showed that ouabain-induced RGC survival depends on autophagy. Using an autophagy inhibitor (3-methyladenine), we observed a complete blockage of the ouabain effect. Western blot analyses showed that ouabain increases the levels of autophagy proteins (LC3 and Beclin-1) coupled to p-CREB transcription factor and leads to autophagosome formation. Additionally, we found that the ratio of cleaved/pro-caspase-3 did not change after ouabain treatment; however, p-JNK density was enhanced. Also, ouabain decreased reactive oxygen species production immediately after axotomy. Taken together, our results suggest that ouabain controls neuroinflammation in the retina following optic nerve axotomy and promotes RGC neuroprotection through activation of the autophagy pathway.
doi_str_mv 10.1007/s11064-021-03481-0
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ispartof Neurochemical research, 2022-03, Vol.47 (3), p.723-738
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1573-6903
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subjects Adenosine Triphosphatases - metabolism
Adenosine Triphosphatases - pharmacology
Animals
Autophagy
Autophagy - physiology
Axotomy
Biochemistry
Biomedical and Life Sciences
Biomedicine
Caspase-3
CD14 antigen
Cell Biology
Cell Survival
Cyclic AMP response element-binding protein
Density
Glial cells
Immunomodulation
Inflammation
Na+/K+-exchanging ATPase
Neonates
Neurochemistry
Neuroinflammatory Diseases
Neurology
Neuronal-glial interactions
Neuroprotection
Neurosciences
Optic nerve
Optic Nerve - physiology
Original Paper
Ouabain
Ouabain - metabolism
Ouabain - pharmacology
Rats
Reactive oxygen species
Reactive Oxygen Species - metabolism
Retina
Retina - metabolism
Retinal cells
Retinal ganglion cells
TLR4 protein
Toll-like receptors
Tumor necrosis factor receptors
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
url http://sfxeu10.hosted.exlibrisgroup.com/loughborough?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-25T14%3A01%3A40IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Ouabain-Na+/K+-ATPase%20Signaling%20Regulates%20Retinal%20Neuroinflammation%20and%20ROS%20Production%20Preventing%20Neuronal%20Death%20by%20an%20Autophagy-Dependent%20Mechanism%20Following%20Optic%20Nerve%20Axotomy%20In%20Vitro&rft.jtitle=Neurochemical%20research&rft.au=M%C3%A1zala-de-Oliveira,%20Thalita&rft.date=2022-03-01&rft.volume=47&rft.issue=3&rft.spage=723&rft.epage=738&rft.pages=723-738&rft.issn=0364-3190&rft.eissn=1573-6903&rft_id=info:doi/10.1007/s11064-021-03481-0&rft_dat=%3Cproquest_cross%3E2628902021%3C/proquest_cross%3E%3Cgrp_id%3Ecdi_FETCH-LOGICAL-c424t-88120f15338be5c901767ea85164dc2f55cac96ab543c37a9aa80a249cc02d663%3C/grp_id%3E%3Coa%3E%3C/oa%3E%3Curl%3E%3C/url%3E&rft_id=info:oai/&rft_pqid=2628902021&rft_id=info:pmid/34783975&rfr_iscdi=true