Oxidative stress and proinflammatory cytokines contribute to demyelination and axonal damage in a cerebellar culture model of neuroinflammation.

<h4>Background</h4>Demyelination and axonal damage are critical processes in the pathogenesis of multiple sclerosis (MS). Oxidative stress and pro-inflammatory cytokines elicited by inflammation mediates tissue damage.<h4>Methods/principal findings</h4>To monitor the demyelin...

Descripción completa

Guardado en:
Detalles Bibliográficos
Autores principales: Alessandra di Penta, Beatriz Moreno, Stephanie Reix, Begoña Fernandez-Diez, Maite Villanueva, Oihana Errea, Nagore Escala, Koen Vandenbroeck, Joan X Comella, Pablo Villoslada
Formato: article
Lenguaje:EN
Publicado: Public Library of Science (PLoS) 2013
Materias:
R
Q
Acceso en línea:https://doaj.org/article/927b9a4c1ce74559acbcc044cb2ad8a1
Etiquetas: Agregar Etiqueta
Sin Etiquetas, Sea el primero en etiquetar este registro!
id oai:doaj.org-article:927b9a4c1ce74559acbcc044cb2ad8a1
record_format dspace
spelling oai:doaj.org-article:927b9a4c1ce74559acbcc044cb2ad8a12021-11-18T07:57:06ZOxidative stress and proinflammatory cytokines contribute to demyelination and axonal damage in a cerebellar culture model of neuroinflammation.1932-620310.1371/journal.pone.0054722https://doaj.org/article/927b9a4c1ce74559acbcc044cb2ad8a12013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23431360/?tool=EBIhttps://doaj.org/toc/1932-6203<h4>Background</h4>Demyelination and axonal damage are critical processes in the pathogenesis of multiple sclerosis (MS). Oxidative stress and pro-inflammatory cytokines elicited by inflammation mediates tissue damage.<h4>Methods/principal findings</h4>To monitor the demyelination and axonal injury associated with microglia activation we employed a model using cerebellar organotypic cultures stimulated with lipopolysaccharide (LPS). Microglia activated by LPS released pro-inflammatory cytokines (IL-1β, IL-6 and TNFα), and increased the expression of inducible nitric oxide synthase (iNOS) and production of reactive oxygen species (ROS). This activation was associated with demyelination and axonal damage in cerebellar cultures. Axonal damage, as revealed by the presence of non-phosphorylated neurofilaments, mitochondrial accumulation in axonal spheroids, and axonal transection, was associated with stronger iNOS expression and concomitant increases in ROS. Moreover, we analyzed the contribution of pro-inflammatory cytokines and oxidative stress in demyelination and axonal degeneration using the iNOS inhibitor ethyl pyruvate, a free-scavenger and xanthine oxidase inhibitor allopurinol, as well as via blockage of pro-inflammatory cytokines using a Fc-TNFR1 construct. We found that blocking microglia activation with ethyl pyruvate or allopurinol significantly decreased axonal damage, and to a lesser extent, demyelination. Blocking TNFα significantly decreased demyelination but did not prevented axonal damage. Moreover, the most common therapy for MS, interferon-beta, was used as an example of an immunomodulator compound that can be tested in this model. In vitro, interferon-beta treatment decreased oxidative stress (iNOS and ROS levels) and the release of pro-inflammatory cytokines after LPS stimulation, reducing axonal damage.<h4>Conclusion</h4>The model of neuroinflammation using cerebellar culture stimulated with endotoxin mimicked myelin and axonal damage mediated by the combination of oxidative stress and pro-inflammatory cytokines. This model may both facilitate understanding of the events involved in neuroinflammation and aid in the development of neuroprotective therapies for the treatment of MS and other neurodegenerative diseases.Alessandra di PentaBeatriz MorenoStephanie ReixBegoña Fernandez-DiezMaite VillanuevaOihana ErreaNagore EscalaKoen VandenbroeckJoan X ComellaPablo VillosladaPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 2, p e54722 (2013)
institution DOAJ
collection DOAJ
language EN
topic Medicine
R
Science
Q
spellingShingle Medicine
R
Science
Q
Alessandra di Penta
Beatriz Moreno
Stephanie Reix
Begoña Fernandez-Diez
Maite Villanueva
Oihana Errea
Nagore Escala
Koen Vandenbroeck
Joan X Comella
Pablo Villoslada
Oxidative stress and proinflammatory cytokines contribute to demyelination and axonal damage in a cerebellar culture model of neuroinflammation.
description <h4>Background</h4>Demyelination and axonal damage are critical processes in the pathogenesis of multiple sclerosis (MS). Oxidative stress and pro-inflammatory cytokines elicited by inflammation mediates tissue damage.<h4>Methods/principal findings</h4>To monitor the demyelination and axonal injury associated with microglia activation we employed a model using cerebellar organotypic cultures stimulated with lipopolysaccharide (LPS). Microglia activated by LPS released pro-inflammatory cytokines (IL-1β, IL-6 and TNFα), and increased the expression of inducible nitric oxide synthase (iNOS) and production of reactive oxygen species (ROS). This activation was associated with demyelination and axonal damage in cerebellar cultures. Axonal damage, as revealed by the presence of non-phosphorylated neurofilaments, mitochondrial accumulation in axonal spheroids, and axonal transection, was associated with stronger iNOS expression and concomitant increases in ROS. Moreover, we analyzed the contribution of pro-inflammatory cytokines and oxidative stress in demyelination and axonal degeneration using the iNOS inhibitor ethyl pyruvate, a free-scavenger and xanthine oxidase inhibitor allopurinol, as well as via blockage of pro-inflammatory cytokines using a Fc-TNFR1 construct. We found that blocking microglia activation with ethyl pyruvate or allopurinol significantly decreased axonal damage, and to a lesser extent, demyelination. Blocking TNFα significantly decreased demyelination but did not prevented axonal damage. Moreover, the most common therapy for MS, interferon-beta, was used as an example of an immunomodulator compound that can be tested in this model. In vitro, interferon-beta treatment decreased oxidative stress (iNOS and ROS levels) and the release of pro-inflammatory cytokines after LPS stimulation, reducing axonal damage.<h4>Conclusion</h4>The model of neuroinflammation using cerebellar culture stimulated with endotoxin mimicked myelin and axonal damage mediated by the combination of oxidative stress and pro-inflammatory cytokines. This model may both facilitate understanding of the events involved in neuroinflammation and aid in the development of neuroprotective therapies for the treatment of MS and other neurodegenerative diseases.
format article
author Alessandra di Penta
Beatriz Moreno
Stephanie Reix
Begoña Fernandez-Diez
Maite Villanueva
Oihana Errea
Nagore Escala
Koen Vandenbroeck
Joan X Comella
Pablo Villoslada
author_facet Alessandra di Penta
Beatriz Moreno
Stephanie Reix
Begoña Fernandez-Diez
Maite Villanueva
Oihana Errea
Nagore Escala
Koen Vandenbroeck
Joan X Comella
Pablo Villoslada
author_sort Alessandra di Penta
title Oxidative stress and proinflammatory cytokines contribute to demyelination and axonal damage in a cerebellar culture model of neuroinflammation.
title_short Oxidative stress and proinflammatory cytokines contribute to demyelination and axonal damage in a cerebellar culture model of neuroinflammation.
title_full Oxidative stress and proinflammatory cytokines contribute to demyelination and axonal damage in a cerebellar culture model of neuroinflammation.
title_fullStr Oxidative stress and proinflammatory cytokines contribute to demyelination and axonal damage in a cerebellar culture model of neuroinflammation.
title_full_unstemmed Oxidative stress and proinflammatory cytokines contribute to demyelination and axonal damage in a cerebellar culture model of neuroinflammation.
title_sort oxidative stress and proinflammatory cytokines contribute to demyelination and axonal damage in a cerebellar culture model of neuroinflammation.
publisher Public Library of Science (PLoS)
publishDate 2013
url https://doaj.org/article/927b9a4c1ce74559acbcc044cb2ad8a1
work_keys_str_mv AT alessandradipenta oxidativestressandproinflammatorycytokinescontributetodemyelinationandaxonaldamageinacerebellarculturemodelofneuroinflammation
AT beatrizmoreno oxidativestressandproinflammatorycytokinescontributetodemyelinationandaxonaldamageinacerebellarculturemodelofneuroinflammation
AT stephaniereix oxidativestressandproinflammatorycytokinescontributetodemyelinationandaxonaldamageinacerebellarculturemodelofneuroinflammation
AT begonafernandezdiez oxidativestressandproinflammatorycytokinescontributetodemyelinationandaxonaldamageinacerebellarculturemodelofneuroinflammation
AT maitevillanueva oxidativestressandproinflammatorycytokinescontributetodemyelinationandaxonaldamageinacerebellarculturemodelofneuroinflammation
AT oihanaerrea oxidativestressandproinflammatorycytokinescontributetodemyelinationandaxonaldamageinacerebellarculturemodelofneuroinflammation
AT nagoreescala oxidativestressandproinflammatorycytokinescontributetodemyelinationandaxonaldamageinacerebellarculturemodelofneuroinflammation
AT koenvandenbroeck oxidativestressandproinflammatorycytokinescontributetodemyelinationandaxonaldamageinacerebellarculturemodelofneuroinflammation
AT joanxcomella oxidativestressandproinflammatorycytokinescontributetodemyelinationandaxonaldamageinacerebellarculturemodelofneuroinflammation
AT pablovilloslada oxidativestressandproinflammatorycytokinescontributetodemyelinationandaxonaldamageinacerebellarculturemodelofneuroinflammation
_version_ 1718422745057853440