Cargando…
Thrombin induces heme oxygenase-1 expression in human synovial fibroblasts through protease-activated receptor signaling pathways
INTRODUCTION: Thrombin is a key factor in the stimulation of fibrin deposition, angiogenesis, and proinflammatory processes. Abnormalities in these processes are primary features of osteoarthritis (OA). Heme oxygenase (HO)-1 is a stress-inducible rate-limiting enzyme in heme degradation that confers...
Autores principales: | , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2012
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3446465/ https://www.ncbi.nlm.nih.gov/pubmed/22541814 http://dx.doi.org/10.1186/ar3815 |
_version_ | 1782243981858439168 |
---|---|
author | Liu, Ju-Fang Hou, Sheng-Mou Tsai, Chun-Hao Huang, Chun-Yin Yang, Wei-Hung Tang, Chih-Hsin |
author_facet | Liu, Ju-Fang Hou, Sheng-Mou Tsai, Chun-Hao Huang, Chun-Yin Yang, Wei-Hung Tang, Chih-Hsin |
author_sort | Liu, Ju-Fang |
collection | PubMed |
description | INTRODUCTION: Thrombin is a key factor in the stimulation of fibrin deposition, angiogenesis, and proinflammatory processes. Abnormalities in these processes are primary features of osteoarthritis (OA). Heme oxygenase (HO)-1 is a stress-inducible rate-limiting enzyme in heme degradation that confers cytoprotection against oxidative injury. Here, we investigated the intracellular signaling pathways involved in thrombin-induced HO-1 expression in human synovial fibroblasts (SFs). METHODS: Thrombin-mediated HO-1 expression was assessed with quantitative real-time (q)PCR. The mechanisms of action of thrombin in different signaling pathways were studied by using Western blotting. Knockdown of protease-activated receptor (PAR) proteins was achieved by transfection with siRNA. Chromatin immunoprecipitation assays were used to study in vivo binding of Nrf2 to the HO-1 promoter. Transient transfection was used to examine HO-1 activity. RESULTS: Osteoarthritis synovial fibroblasts (OASFs) showed significant expression of thrombin, and expression was higher than in normal SFs. OASFs stimulation with thrombin induced concentration- and time-dependent increases in HO-1 expression. Pharmacologic inhibitors or activators and genetic inhibition by siRNA of protease-activated receptors (PARs) revealed that the PAR1 and PAR3 receptors, but not the PAR4 receptor, are involved in thrombin-mediated upregulation of HO-1. Thrombin-mediated HO-1 expression was attenuated by thrombin inhibitor (PPACK), PKCδ inhibitor (rottlerin), or c-Src inhibitor (PP2). Stimulation of cells with thrombin increased PKCδ, c-Src, and Nrf2 activation. CONCLUSION: Our results suggest that the interaction between thrombin and PAR1/PAR3 increases HO-1 expression in human synovial fibroblasts through the PKCδ, c-Src, and Nrf2 signaling pathways. |
format | Online Article Text |
id | pubmed-3446465 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-34464652012-09-20 Thrombin induces heme oxygenase-1 expression in human synovial fibroblasts through protease-activated receptor signaling pathways Liu, Ju-Fang Hou, Sheng-Mou Tsai, Chun-Hao Huang, Chun-Yin Yang, Wei-Hung Tang, Chih-Hsin Arthritis Res Ther Research Article INTRODUCTION: Thrombin is a key factor in the stimulation of fibrin deposition, angiogenesis, and proinflammatory processes. Abnormalities in these processes are primary features of osteoarthritis (OA). Heme oxygenase (HO)-1 is a stress-inducible rate-limiting enzyme in heme degradation that confers cytoprotection against oxidative injury. Here, we investigated the intracellular signaling pathways involved in thrombin-induced HO-1 expression in human synovial fibroblasts (SFs). METHODS: Thrombin-mediated HO-1 expression was assessed with quantitative real-time (q)PCR. The mechanisms of action of thrombin in different signaling pathways were studied by using Western blotting. Knockdown of protease-activated receptor (PAR) proteins was achieved by transfection with siRNA. Chromatin immunoprecipitation assays were used to study in vivo binding of Nrf2 to the HO-1 promoter. Transient transfection was used to examine HO-1 activity. RESULTS: Osteoarthritis synovial fibroblasts (OASFs) showed significant expression of thrombin, and expression was higher than in normal SFs. OASFs stimulation with thrombin induced concentration- and time-dependent increases in HO-1 expression. Pharmacologic inhibitors or activators and genetic inhibition by siRNA of protease-activated receptors (PARs) revealed that the PAR1 and PAR3 receptors, but not the PAR4 receptor, are involved in thrombin-mediated upregulation of HO-1. Thrombin-mediated HO-1 expression was attenuated by thrombin inhibitor (PPACK), PKCδ inhibitor (rottlerin), or c-Src inhibitor (PP2). Stimulation of cells with thrombin increased PKCδ, c-Src, and Nrf2 activation. CONCLUSION: Our results suggest that the interaction between thrombin and PAR1/PAR3 increases HO-1 expression in human synovial fibroblasts through the PKCδ, c-Src, and Nrf2 signaling pathways. BioMed Central 2012 2012-04-27 /pmc/articles/PMC3446465/ /pubmed/22541814 http://dx.doi.org/10.1186/ar3815 Text en Copyright ©2012 Tang et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Article Liu, Ju-Fang Hou, Sheng-Mou Tsai, Chun-Hao Huang, Chun-Yin Yang, Wei-Hung Tang, Chih-Hsin Thrombin induces heme oxygenase-1 expression in human synovial fibroblasts through protease-activated receptor signaling pathways |
title | Thrombin induces heme oxygenase-1 expression in human synovial fibroblasts through protease-activated receptor signaling pathways |
title_full | Thrombin induces heme oxygenase-1 expression in human synovial fibroblasts through protease-activated receptor signaling pathways |
title_fullStr | Thrombin induces heme oxygenase-1 expression in human synovial fibroblasts through protease-activated receptor signaling pathways |
title_full_unstemmed | Thrombin induces heme oxygenase-1 expression in human synovial fibroblasts through protease-activated receptor signaling pathways |
title_short | Thrombin induces heme oxygenase-1 expression in human synovial fibroblasts through protease-activated receptor signaling pathways |
title_sort | thrombin induces heme oxygenase-1 expression in human synovial fibroblasts through protease-activated receptor signaling pathways |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3446465/ https://www.ncbi.nlm.nih.gov/pubmed/22541814 http://dx.doi.org/10.1186/ar3815 |
work_keys_str_mv | AT liujufang thrombininduceshemeoxygenase1expressioninhumansynovialfibroblaststhroughproteaseactivatedreceptorsignalingpathways AT houshengmou thrombininduceshemeoxygenase1expressioninhumansynovialfibroblaststhroughproteaseactivatedreceptorsignalingpathways AT tsaichunhao thrombininduceshemeoxygenase1expressioninhumansynovialfibroblaststhroughproteaseactivatedreceptorsignalingpathways AT huangchunyin thrombininduceshemeoxygenase1expressioninhumansynovialfibroblaststhroughproteaseactivatedreceptorsignalingpathways AT yangweihung thrombininduceshemeoxygenase1expressioninhumansynovialfibroblaststhroughproteaseactivatedreceptorsignalingpathways AT tangchihhsin thrombininduceshemeoxygenase1expressioninhumansynovialfibroblaststhroughproteaseactivatedreceptorsignalingpathways |