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Ciprofloxacin and levofloxacin attenuate microglia inflammatory response via TLR4/NF-kB pathway

BACKGROUND: Neuroinflammation is the response of the central nervous system to events that interfere with tissue homeostasis and represents a common denominator in virtually all neurological diseases. Activation of microglia, the principal immune effector cells of the brain, contributes to neuronal...

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Autores principales: Zusso, Morena, Lunardi, Valentina, Franceschini, Davide, Pagetta, Andrea, Lo, Rita, Stifani, Stefano, Frigo, Anna Chiara, Giusti, Pietro, Moro, Stefano
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6637517/
https://www.ncbi.nlm.nih.gov/pubmed/31319868
http://dx.doi.org/10.1186/s12974-019-1538-9
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author Zusso, Morena
Lunardi, Valentina
Franceschini, Davide
Pagetta, Andrea
Lo, Rita
Stifani, Stefano
Frigo, Anna Chiara
Giusti, Pietro
Moro, Stefano
author_facet Zusso, Morena
Lunardi, Valentina
Franceschini, Davide
Pagetta, Andrea
Lo, Rita
Stifani, Stefano
Frigo, Anna Chiara
Giusti, Pietro
Moro, Stefano
author_sort Zusso, Morena
collection PubMed
description BACKGROUND: Neuroinflammation is the response of the central nervous system to events that interfere with tissue homeostasis and represents a common denominator in virtually all neurological diseases. Activation of microglia, the principal immune effector cells of the brain, contributes to neuronal injury by release of neurotoxic products. Toll-like receptor 4 (TLR4), expressed on the surface of microglia, plays an important role in mediating lipopolysaccharide (LPS)-induced microglia activation and inflammatory responses. We have previously shown that curcumin and some of its analogues harboring an α,β-unsaturated 1,3-diketone moiety, able to coordinate the magnesium ion, can interfere with LPS-mediated TLR4–myeloid differentiation protein-2 (MD-2) signaling. Fluoroquinolone (FQ) antibiotics are compounds that contain a keto-carbonyl group that binds divalent ions, including magnesium. In addition to their antimicrobial activity, FQs are endowed with immunomodulatory properties, but the mechanism underlying their anti-inflammatory activity remains to be defined. The aim of the current study was to elucidate the molecular mechanism of these compounds in the TLR4/NF-κB inflammatory signaling pathway. METHODS: The putative binding mode of five FQs [ciprofloxacin (CPFX), levofloxacin (LVFX), moxifloxacin, ofloxacin, and delafloxacin] to TLR4–MD-2 was determined using molecular docking simulations. The effect of CPFX and LVFX on LPS-induced release of IL-1β and TNF-α and NF-κB activation was investigated in primary microglia by ELISA and fluorescence staining. The interaction of CPFX and LVFX with TLR4–MD-2 complex was assessed by immunoprecipitation followed by Western blotting using Ba/F3 cells. RESULTS: CPFX and LVFX bound to the hydrophobic region of the MD-2 pocket and inhibited LPS-induced secretion of pro-inflammatory cytokines and activation of NF-κB in primary microglia. Furthermore, these FQs diminished the binding of LPS to TLR4–MD-2 complex and decreased the resulting TLR4–MD-2 dimerization in Ba/F3 cells. CONCLUSIONS: These results provide new insight into the mechanism of the anti-inflammatory activity of CPFX and LVFX, which involves, at least in part, the activation of TLR4/NF-κB signaling pathway. Our findings might facilitate the development of new molecules directed at the TLR4–MD-2 complex, a potential key target for controlling neuroinflammation. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12974-019-1538-9) contains supplementary material, which is available to authorized users.
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spelling pubmed-66375172019-07-25 Ciprofloxacin and levofloxacin attenuate microglia inflammatory response via TLR4/NF-kB pathway Zusso, Morena Lunardi, Valentina Franceschini, Davide Pagetta, Andrea Lo, Rita Stifani, Stefano Frigo, Anna Chiara Giusti, Pietro Moro, Stefano J Neuroinflammation Research BACKGROUND: Neuroinflammation is the response of the central nervous system to events that interfere with tissue homeostasis and represents a common denominator in virtually all neurological diseases. Activation of microglia, the principal immune effector cells of the brain, contributes to neuronal injury by release of neurotoxic products. Toll-like receptor 4 (TLR4), expressed on the surface of microglia, plays an important role in mediating lipopolysaccharide (LPS)-induced microglia activation and inflammatory responses. We have previously shown that curcumin and some of its analogues harboring an α,β-unsaturated 1,3-diketone moiety, able to coordinate the magnesium ion, can interfere with LPS-mediated TLR4–myeloid differentiation protein-2 (MD-2) signaling. Fluoroquinolone (FQ) antibiotics are compounds that contain a keto-carbonyl group that binds divalent ions, including magnesium. In addition to their antimicrobial activity, FQs are endowed with immunomodulatory properties, but the mechanism underlying their anti-inflammatory activity remains to be defined. The aim of the current study was to elucidate the molecular mechanism of these compounds in the TLR4/NF-κB inflammatory signaling pathway. METHODS: The putative binding mode of five FQs [ciprofloxacin (CPFX), levofloxacin (LVFX), moxifloxacin, ofloxacin, and delafloxacin] to TLR4–MD-2 was determined using molecular docking simulations. The effect of CPFX and LVFX on LPS-induced release of IL-1β and TNF-α and NF-κB activation was investigated in primary microglia by ELISA and fluorescence staining. The interaction of CPFX and LVFX with TLR4–MD-2 complex was assessed by immunoprecipitation followed by Western blotting using Ba/F3 cells. RESULTS: CPFX and LVFX bound to the hydrophobic region of the MD-2 pocket and inhibited LPS-induced secretion of pro-inflammatory cytokines and activation of NF-κB in primary microglia. Furthermore, these FQs diminished the binding of LPS to TLR4–MD-2 complex and decreased the resulting TLR4–MD-2 dimerization in Ba/F3 cells. CONCLUSIONS: These results provide new insight into the mechanism of the anti-inflammatory activity of CPFX and LVFX, which involves, at least in part, the activation of TLR4/NF-κB signaling pathway. Our findings might facilitate the development of new molecules directed at the TLR4–MD-2 complex, a potential key target for controlling neuroinflammation. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12974-019-1538-9) contains supplementary material, which is available to authorized users. BioMed Central 2019-07-18 /pmc/articles/PMC6637517/ /pubmed/31319868 http://dx.doi.org/10.1186/s12974-019-1538-9 Text en © The Author(s). 2019 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Zusso, Morena
Lunardi, Valentina
Franceschini, Davide
Pagetta, Andrea
Lo, Rita
Stifani, Stefano
Frigo, Anna Chiara
Giusti, Pietro
Moro, Stefano
Ciprofloxacin and levofloxacin attenuate microglia inflammatory response via TLR4/NF-kB pathway
title Ciprofloxacin and levofloxacin attenuate microglia inflammatory response via TLR4/NF-kB pathway
title_full Ciprofloxacin and levofloxacin attenuate microglia inflammatory response via TLR4/NF-kB pathway
title_fullStr Ciprofloxacin and levofloxacin attenuate microglia inflammatory response via TLR4/NF-kB pathway
title_full_unstemmed Ciprofloxacin and levofloxacin attenuate microglia inflammatory response via TLR4/NF-kB pathway
title_short Ciprofloxacin and levofloxacin attenuate microglia inflammatory response via TLR4/NF-kB pathway
title_sort ciprofloxacin and levofloxacin attenuate microglia inflammatory response via tlr4/nf-kb pathway
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6637517/
https://www.ncbi.nlm.nih.gov/pubmed/31319868
http://dx.doi.org/10.1186/s12974-019-1538-9
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