RalA exerts an inhibitory effect on IL-1β/IL-18 secretion by blocking NLRP3 inflammasome activation in levornidazole-treated human THP-1 macrophages.
Animals
Female
Humans
Inflammasomes
/ immunology
Interleukin-18
/ immunology
Interleukin-1beta
/ immunology
Macrophages
/ drug effects
Mice, Inbred C57BL
NLR Family, Pyrin Domain-Containing 3 Protein
/ immunology
Ornidazole
/ pharmacology
RNA, Small Interfering
/ genetics
THP-1 Cells
ral GTP-Binding Proteins
/ genetics
Conformational change
IL-1β/IL-18
NLRP3
RalA
THP-1 macrophages
Journal
International immunopharmacology
ISSN: 1878-1705
Titre abrégé: Int Immunopharmacol
Pays: Netherlands
ID NLM: 100965259
Informations de publication
Date de publication:
Nov 2020
Nov 2020
Historique:
received:
19
06
2020
revised:
11
08
2020
accepted:
11
08
2020
pubmed:
1
9
2020
medline:
28
5
2021
entrez:
1
9
2020
Statut:
ppublish
Résumé
The NLRP3 inflammasome is an important mediator of inflammatory responses and its regulation is an active area of research. RalA is a Ras-like GTPase, which play pivotal roles in the biology of cells. So far, there have been very few studies on RalA regulating inflammatory responses. Bioinformatics analysis predicted that RalA might participate in the regulatory network of NLRP3 inflammasome, which has been confirmed in THP-1 macrophages. After virtual screening of compounds, it was found that levonidazole selected from our virtual small molecule compound library has the potential to bind to RalA. Of note, the interaction of RalA/levornidazole was verified by Surface Plasmon Resonance-Biacore T200, LC/MS analysis and Western blotting analysis. Molecular dynamics simulations revealed that the conformational changes of RalA might be regulated by levornidazole. Additionally, IL-1β/IL-18 secretion from ATP + LPS stimulated THP-1-derived macrophages was RalA-dependently suppressed by levornidazole, suggesting that RalA might have an inhibitory effect on NLRP3 inflammasome activation. The results of co-immunoprecipitation and RalA depletion experiments showed that levornidazole could induce RalA to block the assembly of NLRP3/ASC/pro-caspase-1 complex, thereby reducing the levels of cleaved-caspase-1 and IL-1β/IL-18 secretion. Our study has suggested an anti-inflammatory function of RalA and identified its targeting chemical compound. Overall, this study clarifies a novel pharmacological mechanism by which RalA/levornidazole inhibits NLRP3 inflammasome activation and IL-1β/IL-18 secretion.
Identifiants
pubmed: 32866784
pii: S1567-5769(20)32043-9
doi: 10.1016/j.intimp.2020.106898
pii:
doi:
Substances chimiques
Inflammasomes
0
Interleukin-18
0
Interleukin-1beta
0
NLR Family, Pyrin Domain-Containing 3 Protein
0
RNA, Small Interfering
0
Ornidazole
62XCK0G93T
ral GTP-Binding Proteins
EC 3.6.5.2
levornidazole
H7CGZ9LP53
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
106898Informations de copyright
Copyright © 2020 Elsevier B.V. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.