Evaluation of protein arginine deiminase-4 inhibitor in TNBS- induced colitis in mice.
Animals
Anti-Inflammatory Agents
/ pharmacology
Colitis
/ chemically induced
Colon
/ drug effects
Crohn Disease
/ chemically induced
Cytokines
/ blood
Disease Models, Animal
Extracellular Traps
/ drug effects
Female
Mice, Inbred BALB C
Ornithine
/ analogs & derivatives
Protein-Arginine Deiminase Type 4
/ antagonists & inhibitors
Trinitrobenzenesulfonic Acid
2,4,6-trinitrobenzene sulfonic acid
Cl-amidine
Neutrophil extracellular traps
Protein arginine deiminase-4
Journal
International immunopharmacology
ISSN: 1878-1705
Titre abrégé: Int Immunopharmacol
Pays: Netherlands
ID NLM: 100965259
Informations de publication
Date de publication:
Jul 2020
Jul 2020
Historique:
received:
27
02
2020
revised:
05
05
2020
accepted:
07
05
2020
pubmed:
18
5
2020
medline:
12
3
2021
entrez:
17
5
2020
Statut:
ppublish
Résumé
Many evidences indicated that neutrophil extracellular traps (NETs) are involved in the pathogenesis of inflammatory bowel disease (IBD). Citrullination of histones by Protein Arginine Deiminase-4 (PAD4) is central for NETs formation. This paper aimed to explore the definite role of NETs in mouse model of Crohn's disease (CD) with 2,4,6-trinitrobenzene sulfonic acid (TNBS). The expression of NETs-associated proteins and mRNAs in colon tissue were detected by immunohistochemistry and Real-time Quantitative PCR (QPCR) respectively. Neutrophils were isolated and stimulated in vitro to form NETs. In addition, we also administered Cl-amidine, PAD4 inhibitor, resulting in less NETs formation to investigate protective effect by measuring weight loss, gross bleeding, colon length, myeloperoxidase (MPO) activity, and cytokine expression in mice. The results showed enhanced expression of Ly6G, citrullinated histone H3 (CitH3), and PAD4 in TNBS-induced colitis mice and higher ability of neutrophil to produce NETs in vitro. Blocking NETs formation through Cl-amidine effectively alleviated the clinical colitis index and tissue inflammation in TNBS mice, regulated the expression of pro- or anti-inflammatory cytokines. In addition, Cl-amidine reduced the gene expression of PAD4 and the expression of NETs-associated proteins in the colon of TNBS mice and inhibited the formation of NETs in vitro. Our data showed that Cl-amidine could alleviate the clinical colitis index in TNBS mice to some extend and suggested blocking NETs formation through inhibition of PAD4 as therapeutic targets for the treatment of CD.
Sections du résumé
BACKGROUND AND AIM
OBJECTIVE
Many evidences indicated that neutrophil extracellular traps (NETs) are involved in the pathogenesis of inflammatory bowel disease (IBD). Citrullination of histones by Protein Arginine Deiminase-4 (PAD4) is central for NETs formation. This paper aimed to explore the definite role of NETs in mouse model of Crohn's disease (CD) with 2,4,6-trinitrobenzene sulfonic acid (TNBS).
METHODS
METHODS
The expression of NETs-associated proteins and mRNAs in colon tissue were detected by immunohistochemistry and Real-time Quantitative PCR (QPCR) respectively. Neutrophils were isolated and stimulated in vitro to form NETs. In addition, we also administered Cl-amidine, PAD4 inhibitor, resulting in less NETs formation to investigate protective effect by measuring weight loss, gross bleeding, colon length, myeloperoxidase (MPO) activity, and cytokine expression in mice.
RESULTS
RESULTS
The results showed enhanced expression of Ly6G, citrullinated histone H3 (CitH3), and PAD4 in TNBS-induced colitis mice and higher ability of neutrophil to produce NETs in vitro. Blocking NETs formation through Cl-amidine effectively alleviated the clinical colitis index and tissue inflammation in TNBS mice, regulated the expression of pro- or anti-inflammatory cytokines. In addition, Cl-amidine reduced the gene expression of PAD4 and the expression of NETs-associated proteins in the colon of TNBS mice and inhibited the formation of NETs in vitro.
CONCLUSIONS
CONCLUSIONS
Our data showed that Cl-amidine could alleviate the clinical colitis index in TNBS mice to some extend and suggested blocking NETs formation through inhibition of PAD4 as therapeutic targets for the treatment of CD.
Identifiants
pubmed: 32416455
pii: S1567-5769(20)30543-9
doi: 10.1016/j.intimp.2020.106583
pii:
doi:
Substances chimiques
Anti-Inflammatory Agents
0
Cytokines
0
N-alpha-benzoyl-N5-(2-chloro-1-iminoethyl)-L-ornithine amide
0
Trinitrobenzenesulfonic Acid
8T3HQG2ZC4
Ornithine
E524N2IXA3
Protein-Arginine Deiminase Type 4
EC 3.5.3.15
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
106583Informations 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.