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PYNOD reduces microglial inflammation and consequent neurotoxicity upon lipopolysaccharides stimulation

PYNOD, a nod-like receptors (NLR)-like protein, was indicated to inhibit NF-κB activation, caspase-1-mediated interleukin (IL)-1β release and cell apoptosis in a dose-dependent manner. Exogenous addition of recombinant PYNOD to mixed glial cultures may suppress caspase-1 activation and IL-1β secreti...

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Autores principales: Zeng, Qi, Hu, Chaofeng, Qi, Renbin, Lu, Daxiang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: D.A. Spandidos 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5996706/
https://www.ncbi.nlm.nih.gov/pubmed/29904414
http://dx.doi.org/10.3892/etm.2018.6108
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author Zeng, Qi
Hu, Chaofeng
Qi, Renbin
Lu, Daxiang
author_facet Zeng, Qi
Hu, Chaofeng
Qi, Renbin
Lu, Daxiang
author_sort Zeng, Qi
collection PubMed
description PYNOD, a nod-like receptors (NLR)-like protein, was indicated to inhibit NF-κB activation, caspase-1-mediated interleukin (IL)-1β release and cell apoptosis in a dose-dependent manner. Exogenous addition of recombinant PYNOD to mixed glial cultures may suppress caspase-1 activation and IL-1β secretion induced by Aβ. However, to the best of our knowledge, there no study has focused on the immunoregulatory effects of PYNOD specifically in microglia. The present study aimed to explore the roles of PYNOD involved in the lipopolysaccharides (LPS)-induced microglial inflammation and consequent neurotoxicity. Murine microglial BV-2 cells were transfected with pEGFP-C2-PYNOD (0–5.0 µg/ml) for 24 h and incubated with or without LPS (1 µg/ml) for a further 24 h. Cell viability was determined using MTT assay and the secretion of nitric oxide (NO), IL-1β and caspase-1 was measured using the Griess method or ELISA. Protein expression levels of NF-κB p65 and inducible nitric oxide synthase (iNOS) were detected by immunofluorescent staining and/or western blot analysis. Co-culture of BV-2 cells with human neuroblastoma cell line SK-N-SH was performed in Transwell plates and the cell viability and apoptosis (using flow cytometry) of SK-N-SH cells were determined. Results indicated that PYNOD overexpression inhibited NO secretion and iNOS protein expression induced by LPS in BV-2 cells, with no detectable cytotoxicity. PYNOD overexpression also reduced the secretion of IL-1β and caspase-1 from BV-2 cells upon LPS stimulation. These effects were dose-dependent. Additionally, PYNOD overexpression prevented LPS-induced nuclear translocation of NF-κB p65 in BV-2 cells. The growth-inhibitory and apoptosis-promoting effects of BV-2 cells towards SK-N-SH cells were alleviated as a result of PYNOD overexpression. In conclusion, PYNOD may mitigate microglial inflammation and consequent neurotoxicity.
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spelling pubmed-59967062018-06-14 PYNOD reduces microglial inflammation and consequent neurotoxicity upon lipopolysaccharides stimulation Zeng, Qi Hu, Chaofeng Qi, Renbin Lu, Daxiang Exp Ther Med Articles PYNOD, a nod-like receptors (NLR)-like protein, was indicated to inhibit NF-κB activation, caspase-1-mediated interleukin (IL)-1β release and cell apoptosis in a dose-dependent manner. Exogenous addition of recombinant PYNOD to mixed glial cultures may suppress caspase-1 activation and IL-1β secretion induced by Aβ. However, to the best of our knowledge, there no study has focused on the immunoregulatory effects of PYNOD specifically in microglia. The present study aimed to explore the roles of PYNOD involved in the lipopolysaccharides (LPS)-induced microglial inflammation and consequent neurotoxicity. Murine microglial BV-2 cells were transfected with pEGFP-C2-PYNOD (0–5.0 µg/ml) for 24 h and incubated with or without LPS (1 µg/ml) for a further 24 h. Cell viability was determined using MTT assay and the secretion of nitric oxide (NO), IL-1β and caspase-1 was measured using the Griess method or ELISA. Protein expression levels of NF-κB p65 and inducible nitric oxide synthase (iNOS) were detected by immunofluorescent staining and/or western blot analysis. Co-culture of BV-2 cells with human neuroblastoma cell line SK-N-SH was performed in Transwell plates and the cell viability and apoptosis (using flow cytometry) of SK-N-SH cells were determined. Results indicated that PYNOD overexpression inhibited NO secretion and iNOS protein expression induced by LPS in BV-2 cells, with no detectable cytotoxicity. PYNOD overexpression also reduced the secretion of IL-1β and caspase-1 from BV-2 cells upon LPS stimulation. These effects were dose-dependent. Additionally, PYNOD overexpression prevented LPS-induced nuclear translocation of NF-κB p65 in BV-2 cells. The growth-inhibitory and apoptosis-promoting effects of BV-2 cells towards SK-N-SH cells were alleviated as a result of PYNOD overexpression. In conclusion, PYNOD may mitigate microglial inflammation and consequent neurotoxicity. D.A. Spandidos 2018-06 2018-04-30 /pmc/articles/PMC5996706/ /pubmed/29904414 http://dx.doi.org/10.3892/etm.2018.6108 Text en Copyright: © Zeng et al. This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License (https://creativecommons.org/licenses/by-nc-nd/4.0/) , which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.
spellingShingle Articles
Zeng, Qi
Hu, Chaofeng
Qi, Renbin
Lu, Daxiang
PYNOD reduces microglial inflammation and consequent neurotoxicity upon lipopolysaccharides stimulation
title PYNOD reduces microglial inflammation and consequent neurotoxicity upon lipopolysaccharides stimulation
title_full PYNOD reduces microglial inflammation and consequent neurotoxicity upon lipopolysaccharides stimulation
title_fullStr PYNOD reduces microglial inflammation and consequent neurotoxicity upon lipopolysaccharides stimulation
title_full_unstemmed PYNOD reduces microglial inflammation and consequent neurotoxicity upon lipopolysaccharides stimulation
title_short PYNOD reduces microglial inflammation and consequent neurotoxicity upon lipopolysaccharides stimulation
title_sort pynod reduces microglial inflammation and consequent neurotoxicity upon lipopolysaccharides stimulation
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5996706/
https://www.ncbi.nlm.nih.gov/pubmed/29904414
http://dx.doi.org/10.3892/etm.2018.6108
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