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Lipopolysaccharide pretreatment increases protease-activated receptor-2 expression and monocyte chemoattractant protein-1 secretion in vascular endothelial cells

BACKGROUND: This study investigated whether lipopolysaccharide (LPS) increase protease-activated receptor-2 (PAR-2) expression and enhance the association between PAR-2 expression and chemokine production in human vascular endothelial cells (ECs). METHODS: The morphology of ECs was observed through...

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Autores principales: Chao, Hung-Hsing, Chen, Po-Yuan, Hao, Wen-Rui, Chiang, Wei-Ping, Cheng, Tzu-Hurng, Loh, Shih-Hurng, Leung, Yuk-Man, Liu, Ju-Chi, Chen, Jin-Jer, Sung, Li-Chin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5688698/
https://www.ncbi.nlm.nih.gov/pubmed/29141644
http://dx.doi.org/10.1186/s12929-017-0393-1
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author Chao, Hung-Hsing
Chen, Po-Yuan
Hao, Wen-Rui
Chiang, Wei-Ping
Cheng, Tzu-Hurng
Loh, Shih-Hurng
Leung, Yuk-Man
Liu, Ju-Chi
Chen, Jin-Jer
Sung, Li-Chin
author_facet Chao, Hung-Hsing
Chen, Po-Yuan
Hao, Wen-Rui
Chiang, Wei-Ping
Cheng, Tzu-Hurng
Loh, Shih-Hurng
Leung, Yuk-Man
Liu, Ju-Chi
Chen, Jin-Jer
Sung, Li-Chin
author_sort Chao, Hung-Hsing
collection PubMed
description BACKGROUND: This study investigated whether lipopolysaccharide (LPS) increase protease-activated receptor-2 (PAR-2) expression and enhance the association between PAR-2 expression and chemokine production in human vascular endothelial cells (ECs). METHODS: The morphology of ECs was observed through microphotography in cultured human umbilical vein ECs (EA. hy926 cells) treated with various LPS concentrations (0, 0.25, 0.5, 1, and 2 μg/mL) for 24 h, and cell viability was assessed using the MTT assay. Intracellular calcium imaging was performed to assess agonist (trypsin)-induced PAR-2 activity. Western blotting was used to explore the LPS-mediated signal transduction pathway and the expression of PAR-2 and adhesion molecule monocyte chemoattractant protein-1 (MCP-1) in ECs. RESULTS: Trypsin stimulation increased intracellular calcium release in ECs. The calcium influx was augmented in cells pretreated with a high LPS concentration (1 μg/mL). After 24 h treatment of LPS, no changes in ECs viability or morphology were observed. Western blotting revealed that LPS increased PAR-2 expression and enhanced trypsin-induced extracellular signal-regulated kinase (ERK)/p38 phosphorylation and MCP-1 secretion. However, pretreatment with selective ERK (PD98059), p38 mitogen-activated protein kinase (MAPK) (SB203580) inhibitors, and the selective PAR-2 antagonist (FSLLRY-NH2) blocked the effects of LPS-activated PAR-2 on MCP-1 secretion. CONCLUSIONS: Our findings provide the first evidence that the bacterial endotoxin LPS potentiates calcium mobilization and ERK/p38 MAPK pathway activation and leads to the secretion of the pro-inflammatory chemokine MCP-1 by inducing PAR-2 expression and its associated activity in vascular ECs. Therefore, PAR-2 exerts vascular inflammatory effects and plays an important role in bacterial infection-induced pathological responses. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12929-017-0393-1) contains supplementary material, which is available to authorized users.
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spelling pubmed-56886982017-11-24 Lipopolysaccharide pretreatment increases protease-activated receptor-2 expression and monocyte chemoattractant protein-1 secretion in vascular endothelial cells Chao, Hung-Hsing Chen, Po-Yuan Hao, Wen-Rui Chiang, Wei-Ping Cheng, Tzu-Hurng Loh, Shih-Hurng Leung, Yuk-Man Liu, Ju-Chi Chen, Jin-Jer Sung, Li-Chin J Biomed Sci Research BACKGROUND: This study investigated whether lipopolysaccharide (LPS) increase protease-activated receptor-2 (PAR-2) expression and enhance the association between PAR-2 expression and chemokine production in human vascular endothelial cells (ECs). METHODS: The morphology of ECs was observed through microphotography in cultured human umbilical vein ECs (EA. hy926 cells) treated with various LPS concentrations (0, 0.25, 0.5, 1, and 2 μg/mL) for 24 h, and cell viability was assessed using the MTT assay. Intracellular calcium imaging was performed to assess agonist (trypsin)-induced PAR-2 activity. Western blotting was used to explore the LPS-mediated signal transduction pathway and the expression of PAR-2 and adhesion molecule monocyte chemoattractant protein-1 (MCP-1) in ECs. RESULTS: Trypsin stimulation increased intracellular calcium release in ECs. The calcium influx was augmented in cells pretreated with a high LPS concentration (1 μg/mL). After 24 h treatment of LPS, no changes in ECs viability or morphology were observed. Western blotting revealed that LPS increased PAR-2 expression and enhanced trypsin-induced extracellular signal-regulated kinase (ERK)/p38 phosphorylation and MCP-1 secretion. However, pretreatment with selective ERK (PD98059), p38 mitogen-activated protein kinase (MAPK) (SB203580) inhibitors, and the selective PAR-2 antagonist (FSLLRY-NH2) blocked the effects of LPS-activated PAR-2 on MCP-1 secretion. CONCLUSIONS: Our findings provide the first evidence that the bacterial endotoxin LPS potentiates calcium mobilization and ERK/p38 MAPK pathway activation and leads to the secretion of the pro-inflammatory chemokine MCP-1 by inducing PAR-2 expression and its associated activity in vascular ECs. Therefore, PAR-2 exerts vascular inflammatory effects and plays an important role in bacterial infection-induced pathological responses. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12929-017-0393-1) contains supplementary material, which is available to authorized users. BioMed Central 2017-11-15 /pmc/articles/PMC5688698/ /pubmed/29141644 http://dx.doi.org/10.1186/s12929-017-0393-1 Text en © The Author(s). 2017 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
Chao, Hung-Hsing
Chen, Po-Yuan
Hao, Wen-Rui
Chiang, Wei-Ping
Cheng, Tzu-Hurng
Loh, Shih-Hurng
Leung, Yuk-Man
Liu, Ju-Chi
Chen, Jin-Jer
Sung, Li-Chin
Lipopolysaccharide pretreatment increases protease-activated receptor-2 expression and monocyte chemoattractant protein-1 secretion in vascular endothelial cells
title Lipopolysaccharide pretreatment increases protease-activated receptor-2 expression and monocyte chemoattractant protein-1 secretion in vascular endothelial cells
title_full Lipopolysaccharide pretreatment increases protease-activated receptor-2 expression and monocyte chemoattractant protein-1 secretion in vascular endothelial cells
title_fullStr Lipopolysaccharide pretreatment increases protease-activated receptor-2 expression and monocyte chemoattractant protein-1 secretion in vascular endothelial cells
title_full_unstemmed Lipopolysaccharide pretreatment increases protease-activated receptor-2 expression and monocyte chemoattractant protein-1 secretion in vascular endothelial cells
title_short Lipopolysaccharide pretreatment increases protease-activated receptor-2 expression and monocyte chemoattractant protein-1 secretion in vascular endothelial cells
title_sort lipopolysaccharide pretreatment increases protease-activated receptor-2 expression and monocyte chemoattractant protein-1 secretion in vascular endothelial cells
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5688698/
https://www.ncbi.nlm.nih.gov/pubmed/29141644
http://dx.doi.org/10.1186/s12929-017-0393-1
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