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NOD-Like Receptor Protein 3 Inflammasome Priming and Activation in Barrett’s Epithelial Cells

BACKGROUND & AIMS: Microbial molecular products incite intestinal inflammation by activating Toll-like receptors (TLRs) and inflammasomes of the innate immune system. This system’s contribution to esophageal inflammation is not known. Gram-negative bacteria, which dominate the esophageal microbi...

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Autores principales: Nadatani, Yuji, Huo, Xiaofang, Zhang, Xi, Yu, Chunhua, Cheng, Edaire, Zhang, Qiuyang, Dunbar, Kerry B., Theiss, Arianne, Pham, Thai H., Wang, David H., Watanabe, Toshio, Fujiwara, Yasuhiro, Arakawa, Tetsuo, Spechler, Stuart J., Souza, Rhonda F.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5042564/
https://www.ncbi.nlm.nih.gov/pubmed/27777967
http://dx.doi.org/10.1016/j.jcmgh.2016.03.006
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author Nadatani, Yuji
Huo, Xiaofang
Zhang, Xi
Yu, Chunhua
Cheng, Edaire
Zhang, Qiuyang
Dunbar, Kerry B.
Theiss, Arianne
Pham, Thai H.
Wang, David H.
Watanabe, Toshio
Fujiwara, Yasuhiro
Arakawa, Tetsuo
Spechler, Stuart J.
Souza, Rhonda F.
author_facet Nadatani, Yuji
Huo, Xiaofang
Zhang, Xi
Yu, Chunhua
Cheng, Edaire
Zhang, Qiuyang
Dunbar, Kerry B.
Theiss, Arianne
Pham, Thai H.
Wang, David H.
Watanabe, Toshio
Fujiwara, Yasuhiro
Arakawa, Tetsuo
Spechler, Stuart J.
Souza, Rhonda F.
author_sort Nadatani, Yuji
collection PubMed
description BACKGROUND & AIMS: Microbial molecular products incite intestinal inflammation by activating Toll-like receptors (TLRs) and inflammasomes of the innate immune system. This system’s contribution to esophageal inflammation is not known. Gram-negative bacteria, which dominate the esophageal microbiome in reflux esophagitis, produce lipopolysaccharide (LPS), a TLR4 ligand. TLR4 signaling produces pro-interleukin (IL)1β, pro-IL18, and NOD-like receptor protein 3 (NLRP3), which prime the NLRP3 inflammasome. Subsequent NLRP3 inflammasome activation cleaves caspase-1, inducing secretion of proinflammatory cytokines and pyroptosis (inflammatory cell death). We explored LPS effects on NLRP3 inflammasome priming and activation in esophageal cells. METHODS: We exposed esophageal squamous and Barrett’s epithelial cells to LPS and measured the following: (1) TLR4, pro-IL1β, pro-IL18, and NLRP3 expression; (2) caspase-1 activity; (3) tumor necrosis factor-α, IL8, IL1β, and IL18 secretion; (4) lactate dehydrogenase (LDH) release (a pyroptosis marker); and (5) mitochondrial reactive oxygen species (ROS). As inhibitors, we used acetyl-Tyr-Val-Ala-Asp-CHO for caspase-1, small interfering RNA for NLRP3, and (2-(2,2,6,6,-Tetramethylpiperidin-1-oxyl-4-ylamino)-2-oxoethyl)triphenylphosphonium chloride for mitochondrial ROS. RESULTS: Squamous and Barrett’s cells expressed similar levels of TLR4, but LPS induced TLR4 signaling that increased tumor necrosis factor-α and IL8 secretion only in Barrett’s cells. Barrett’s cells treated with LPS showed increased expression of pro-IL18, pro-IL1β, and NLRP3, and increased mitochondrial ROS levels, caspase-1 activity, IL1β and IL18 secretion, and LDH release. Acetyl-Tyr-Val-Ala-Asp-CHO, NLRP3 small interfering RNA, and Mito-TEMPO all blocked LPS-induced IL1β and IL18 secretion and LDH release. CONCLUSIONS: In Barrett’s cells, LPS both primes and activates the NLRP3 inflammasome, causing secretion of proinflammatory cytokines and pyroptosis. By triggering molecular events promoting inflammation, the esophageal microbiome might contribute to inflammation-mediated carcinogenesis in Barrett’s esophagus.
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spelling pubmed-50425642016-12-15 NOD-Like Receptor Protein 3 Inflammasome Priming and Activation in Barrett’s Epithelial Cells Nadatani, Yuji Huo, Xiaofang Zhang, Xi Yu, Chunhua Cheng, Edaire Zhang, Qiuyang Dunbar, Kerry B. Theiss, Arianne Pham, Thai H. Wang, David H. Watanabe, Toshio Fujiwara, Yasuhiro Arakawa, Tetsuo Spechler, Stuart J. Souza, Rhonda F. Cell Mol Gastroenterol Hepatol Original Research BACKGROUND & AIMS: Microbial molecular products incite intestinal inflammation by activating Toll-like receptors (TLRs) and inflammasomes of the innate immune system. This system’s contribution to esophageal inflammation is not known. Gram-negative bacteria, which dominate the esophageal microbiome in reflux esophagitis, produce lipopolysaccharide (LPS), a TLR4 ligand. TLR4 signaling produces pro-interleukin (IL)1β, pro-IL18, and NOD-like receptor protein 3 (NLRP3), which prime the NLRP3 inflammasome. Subsequent NLRP3 inflammasome activation cleaves caspase-1, inducing secretion of proinflammatory cytokines and pyroptosis (inflammatory cell death). We explored LPS effects on NLRP3 inflammasome priming and activation in esophageal cells. METHODS: We exposed esophageal squamous and Barrett’s epithelial cells to LPS and measured the following: (1) TLR4, pro-IL1β, pro-IL18, and NLRP3 expression; (2) caspase-1 activity; (3) tumor necrosis factor-α, IL8, IL1β, and IL18 secretion; (4) lactate dehydrogenase (LDH) release (a pyroptosis marker); and (5) mitochondrial reactive oxygen species (ROS). As inhibitors, we used acetyl-Tyr-Val-Ala-Asp-CHO for caspase-1, small interfering RNA for NLRP3, and (2-(2,2,6,6,-Tetramethylpiperidin-1-oxyl-4-ylamino)-2-oxoethyl)triphenylphosphonium chloride for mitochondrial ROS. RESULTS: Squamous and Barrett’s cells expressed similar levels of TLR4, but LPS induced TLR4 signaling that increased tumor necrosis factor-α and IL8 secretion only in Barrett’s cells. Barrett’s cells treated with LPS showed increased expression of pro-IL18, pro-IL1β, and NLRP3, and increased mitochondrial ROS levels, caspase-1 activity, IL1β and IL18 secretion, and LDH release. Acetyl-Tyr-Val-Ala-Asp-CHO, NLRP3 small interfering RNA, and Mito-TEMPO all blocked LPS-induced IL1β and IL18 secretion and LDH release. CONCLUSIONS: In Barrett’s cells, LPS both primes and activates the NLRP3 inflammasome, causing secretion of proinflammatory cytokines and pyroptosis. By triggering molecular events promoting inflammation, the esophageal microbiome might contribute to inflammation-mediated carcinogenesis in Barrett’s esophagus. Elsevier 2016-03-19 /pmc/articles/PMC5042564/ /pubmed/27777967 http://dx.doi.org/10.1016/j.jcmgh.2016.03.006 Text en © 2016 The Authors http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Original Research
Nadatani, Yuji
Huo, Xiaofang
Zhang, Xi
Yu, Chunhua
Cheng, Edaire
Zhang, Qiuyang
Dunbar, Kerry B.
Theiss, Arianne
Pham, Thai H.
Wang, David H.
Watanabe, Toshio
Fujiwara, Yasuhiro
Arakawa, Tetsuo
Spechler, Stuart J.
Souza, Rhonda F.
NOD-Like Receptor Protein 3 Inflammasome Priming and Activation in Barrett’s Epithelial Cells
title NOD-Like Receptor Protein 3 Inflammasome Priming and Activation in Barrett’s Epithelial Cells
title_full NOD-Like Receptor Protein 3 Inflammasome Priming and Activation in Barrett’s Epithelial Cells
title_fullStr NOD-Like Receptor Protein 3 Inflammasome Priming and Activation in Barrett’s Epithelial Cells
title_full_unstemmed NOD-Like Receptor Protein 3 Inflammasome Priming and Activation in Barrett’s Epithelial Cells
title_short NOD-Like Receptor Protein 3 Inflammasome Priming and Activation in Barrett’s Epithelial Cells
title_sort nod-like receptor protein 3 inflammasome priming and activation in barrett’s epithelial cells
topic Original Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5042564/
https://www.ncbi.nlm.nih.gov/pubmed/27777967
http://dx.doi.org/10.1016/j.jcmgh.2016.03.006
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