di Penta A.,Institute of Biomedical Research August Pi Sunyer IDIBAPS Hospital Clinic of Barcelona |
di Penta A.,University of the Basque Country |
Moreno B.,Institute of Biomedical Research August Pi Sunyer IDIBAPS Hospital Clinic of Barcelona |
Reix S.,Autonomous University of Barcelona |
And 8 more authors.
PLoS ONE | Year: 2013
Background: 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. Methods/Principal Findings: 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. Conclusion: 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. © 2013 di Penta et al.
PubMed | Juntendo University, Institute of Biomedical Research August Pi Sunyer IDIBAPS Hospital Clinic of Barcelona, University of the Basque Country and National Institute of Neuroscience
Type: Journal Article | Journal: PloS one | Year: 2013
Celecoxib is a selective cyclooxygenase-2 (COX2) inhibitor. We have previously shown that celecoxib inhibits experimental autoimmune encephalomyelitis (EAE) in COX-2-deficient mice, suggestive for a mode of action involving COX2-independent pathways. In the present study, we tested the effect of a trifluoromethyl analogue of celecoxib (TFM-C) with 205-fold lower COX-2 inhibitory activity in two models of neuroinflammation, i.e. cerebellar organotypic cultures challenged with LPS and the EAE mouse model for multiple sclerosis. TFM-C inhibited secretion of IL-1, IL-12 and IL-17, enhanced that of TNF- and RANTES, reduced neuronal axonal damage and protected from oxidative stress in the organotypic model. TFM-C blocked TNF- release in microglial cells through a process involving intracellular retention, but induced TNF- secretion in primary astrocyte cultures. Finally, we demonstrate that TFM-C and celecoxib ameliorated EAE with equal potency. This coincided with reduced secretion of IL-17 and IFN- by MOG-reactive T-cells and of IL-23 and inflammatory cytokines by bone marrow-derived dendritic cells. Our study reveals that non-coxib analogues of celecoxib may have translational value in the treatment of neuro-inflammatory conditions.
PubMed | University of Navarra, Institute of Biomedical Research August Pi Sunyer IDIBAPS Hospital Clinic of Barcelona and Navarra Health Service Hospital
Type: | Journal: Journal of attention disorders | Year: 2015
Group A Streptococcus has been associated with ADHD, tic disorders (TD), and obsessive-compulsive disorder (OCD) through anti-basal ganglia antibodies (ABGA).We investigated the association between ABGA and streptococcal exposure with behavioral, motor, and cognitive measures in 38 children with ADHD not comorbid to OCD or TD (nc-ADHD) and in 38 healthy children. An additional group of 15 children with TD and/or OCD was examined.ABGA titers were present in 3% of nc-ADHD patients and controls but in 27% of TD and/or OCD patients. Evidence of streptococcal exposure was similar between ADHD patients and controls living in the same urban area. Behavioral, motor, and cognitive measures were not associated with anti-streptococcal antibodies.ABGA do not distinguish nc-ADHD from controls. The differences in the frequency of streptococcal exposure in previous studies are determined by the dynamic nature of the infection rather than the behavioral phenotype of ADHD.