Neuroprotective Effect of Neuroserpin in Oxygen-Glucose Deprivation- and Reoxygenation-Treated Rat Astrocytes In Vitro

Neuroserpin (NSP) reportedly exerts neuroprotective effects in cerebral ischemic animal models and patients; however, the mechanism of protection is poorly understood. We thus attempted to confirm neuroprotective effects of NSP on astrocytes in the ischemic state and then explored the relative mecha...

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Autores principales: Wang, Liang, Zhang, Yang, Asakawa, Tetsuya, Li, Wei, Han, Sha, Li, Qinying, Xiao, Baoguo, Namba, Hiroki, Lu, Chuanzhen, Dong, Qiang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4395230/
https://www.ncbi.nlm.nih.gov/pubmed/25874935
http://dx.doi.org/10.1371/journal.pone.0123932
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author Wang, Liang
Zhang, Yang
Asakawa, Tetsuya
Li, Wei
Han, Sha
Li, Qinying
Xiao, Baoguo
Namba, Hiroki
Lu, Chuanzhen
Dong, Qiang
author_facet Wang, Liang
Zhang, Yang
Asakawa, Tetsuya
Li, Wei
Han, Sha
Li, Qinying
Xiao, Baoguo
Namba, Hiroki
Lu, Chuanzhen
Dong, Qiang
author_sort Wang, Liang
collection PubMed
description Neuroserpin (NSP) reportedly exerts neuroprotective effects in cerebral ischemic animal models and patients; however, the mechanism of protection is poorly understood. We thus attempted to confirm neuroprotective effects of NSP on astrocytes in the ischemic state and then explored the relative mechanisms. Astrocytes from neonatal rats were treated with oxygen-glucose deprivation (OGD) followed by reoxygenation (OGD/R). To confirm the neuroprotective effects of NSP, we measured the cell survival rate, relative lactate dehydrogenase (LDH) release; we also performed morphological methods, namely Hoechst 33342 staining and Annexin V assay. To explore the potential mechanisms of NSP, the release of nitric oxide (NO) and TNF-α related to NSP administration were measured by enzyme-linked immunosorbent assay. The proteins related to the NF-κB, ERK1/2, and PI3K/Akt pathways were investigated by Western blotting. To verify the cause-and-effect relationship between neuroprotection and the NF-κB pathway, a NF-κB pathway inhibitor sc3060 was employed to observe the effects of NSP-induced neuroprotection. We found that NSP significantly increased the cell survival rate and reduced LDH release in OGD/R-treated astrocytes. It also reduced NO/TNF-α release. Western blotting showed that the protein levels of p-IKKBα/β and P65 were upregulated by the OGD/R treatment and such effects were significantly inhibited by NSP administration. The NSP-induced inhibition could be significantly reversed by administration of the NF-κB pathway inhibitor sc3060, whereas, expressions of p-ERK1, p-ERK2, and p-AKT were upregulated by the OGD/R treatment; however, their levels were unchanged by NSP administration. Our results thus verified the neuroprotective effects of NSP in ischemic astrocytes. The potential mechanisms include inhibition of the release of NO/TNF-α and repression of the NF-κB signaling pathways. Our data also indicated that NSP has little influence on the MAPK and PI3K/Akt pathways.
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spelling pubmed-43952302015-04-21 Neuroprotective Effect of Neuroserpin in Oxygen-Glucose Deprivation- and Reoxygenation-Treated Rat Astrocytes In Vitro Wang, Liang Zhang, Yang Asakawa, Tetsuya Li, Wei Han, Sha Li, Qinying Xiao, Baoguo Namba, Hiroki Lu, Chuanzhen Dong, Qiang PLoS One Research Article Neuroserpin (NSP) reportedly exerts neuroprotective effects in cerebral ischemic animal models and patients; however, the mechanism of protection is poorly understood. We thus attempted to confirm neuroprotective effects of NSP on astrocytes in the ischemic state and then explored the relative mechanisms. Astrocytes from neonatal rats were treated with oxygen-glucose deprivation (OGD) followed by reoxygenation (OGD/R). To confirm the neuroprotective effects of NSP, we measured the cell survival rate, relative lactate dehydrogenase (LDH) release; we also performed morphological methods, namely Hoechst 33342 staining and Annexin V assay. To explore the potential mechanisms of NSP, the release of nitric oxide (NO) and TNF-α related to NSP administration were measured by enzyme-linked immunosorbent assay. The proteins related to the NF-κB, ERK1/2, and PI3K/Akt pathways were investigated by Western blotting. To verify the cause-and-effect relationship between neuroprotection and the NF-κB pathway, a NF-κB pathway inhibitor sc3060 was employed to observe the effects of NSP-induced neuroprotection. We found that NSP significantly increased the cell survival rate and reduced LDH release in OGD/R-treated astrocytes. It also reduced NO/TNF-α release. Western blotting showed that the protein levels of p-IKKBα/β and P65 were upregulated by the OGD/R treatment and such effects were significantly inhibited by NSP administration. The NSP-induced inhibition could be significantly reversed by administration of the NF-κB pathway inhibitor sc3060, whereas, expressions of p-ERK1, p-ERK2, and p-AKT were upregulated by the OGD/R treatment; however, their levels were unchanged by NSP administration. Our results thus verified the neuroprotective effects of NSP in ischemic astrocytes. The potential mechanisms include inhibition of the release of NO/TNF-α and repression of the NF-κB signaling pathways. Our data also indicated that NSP has little influence on the MAPK and PI3K/Akt pathways. Public Library of Science 2015-04-13 /pmc/articles/PMC4395230/ /pubmed/25874935 http://dx.doi.org/10.1371/journal.pone.0123932 Text en © 2015 Wang et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Wang, Liang
Zhang, Yang
Asakawa, Tetsuya
Li, Wei
Han, Sha
Li, Qinying
Xiao, Baoguo
Namba, Hiroki
Lu, Chuanzhen
Dong, Qiang
Neuroprotective Effect of Neuroserpin in Oxygen-Glucose Deprivation- and Reoxygenation-Treated Rat Astrocytes In Vitro
title Neuroprotective Effect of Neuroserpin in Oxygen-Glucose Deprivation- and Reoxygenation-Treated Rat Astrocytes In Vitro
title_full Neuroprotective Effect of Neuroserpin in Oxygen-Glucose Deprivation- and Reoxygenation-Treated Rat Astrocytes In Vitro
title_fullStr Neuroprotective Effect of Neuroserpin in Oxygen-Glucose Deprivation- and Reoxygenation-Treated Rat Astrocytes In Vitro
title_full_unstemmed Neuroprotective Effect of Neuroserpin in Oxygen-Glucose Deprivation- and Reoxygenation-Treated Rat Astrocytes In Vitro
title_short Neuroprotective Effect of Neuroserpin in Oxygen-Glucose Deprivation- and Reoxygenation-Treated Rat Astrocytes In Vitro
title_sort neuroprotective effect of neuroserpin in oxygen-glucose deprivation- and reoxygenation-treated rat astrocytes in vitro
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4395230/
https://www.ncbi.nlm.nih.gov/pubmed/25874935
http://dx.doi.org/10.1371/journal.pone.0123932
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