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Thrombin-Induced COX-2 Expression and PGE(2) Synthesis in Human Tracheal Smooth Muscle Cells: Role of PKCδ/Pyk2-Dependent AP-1 Pathway Modulation

In this study, we confirmed that thrombin significantly increases the production of COX-2 and PGE(2) in human tracheal smooth muscle cells (HTSMCs), leading to inflammation in the airways and lungs. These molecules are well-known contributors to various inflammatory diseases. Here, we investigated i...

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Autores principales: Yang, Chien-Chung, Lee, I-Ta, Lin, Yan-Jyun, Wu, Wen-Bin, Hsiao, Li-Der, Yang, Chuen-Mao
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10606820/
https://www.ncbi.nlm.nih.gov/pubmed/37894811
http://dx.doi.org/10.3390/ijms242015130
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author Yang, Chien-Chung
Lee, I-Ta
Lin, Yan-Jyun
Wu, Wen-Bin
Hsiao, Li-Der
Yang, Chuen-Mao
author_facet Yang, Chien-Chung
Lee, I-Ta
Lin, Yan-Jyun
Wu, Wen-Bin
Hsiao, Li-Der
Yang, Chuen-Mao
author_sort Yang, Chien-Chung
collection PubMed
description In this study, we confirmed that thrombin significantly increases the production of COX-2 and PGE(2) in human tracheal smooth muscle cells (HTSMCs), leading to inflammation in the airways and lungs. These molecules are well-known contributors to various inflammatory diseases. Here, we investigated in detail the involved signaling pathways using specific inhibitors and small interfering RNAs (siRNAs). Our results demonstrated that inhibitors targeting proteins such as protein kinase C (PKC)δ, proline-rich tyrosine kinase 2 (Pyk2), c-Src, epidermal growth factor receptor (EGFR), phosphatidylinositol 3-kinase (PI3K), or activator protein-1 (AP-1) effectively reduced thrombin-induced COX-2 and PGE(2) production. Additionally, transfection with siRNAs against PKCδ, Pyk2, c-Src, EGFR, protein kinase B (Akt), or c-Jun mitigated these responses. Furthermore, our observations revealed that thrombin stimulated the phosphorylation of key components of the signaling cascade, including PKCδ, Pyk2, c-Src, EGFR, Akt, and c-Jun. Thrombin activated COX-2 promoter activity through AP-1 activation, a process that was disrupted by a point-mutated AP-1 site within the COX-2 promoter. Finally, resveratrol (one of the most researched natural polyphenols) was found to effectively inhibit thrombin-induced COX-2 expression and PGE(2) release in HTSMCs through blocking the activation of Pyk2, c-Src, EGFR, Akt, and c-Jun. In summary, our findings demonstrate that thrombin-induced COX-2 and PGE(2) generation involves a PKCδ/Pyk2/c-Src/EGFR/PI3K/Akt-dependent AP-1 activation pathway. This study also suggests the potential use of resveratrol as an intervention for managing airway inflammation.
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spelling pubmed-106068202023-10-28 Thrombin-Induced COX-2 Expression and PGE(2) Synthesis in Human Tracheal Smooth Muscle Cells: Role of PKCδ/Pyk2-Dependent AP-1 Pathway Modulation Yang, Chien-Chung Lee, I-Ta Lin, Yan-Jyun Wu, Wen-Bin Hsiao, Li-Der Yang, Chuen-Mao Int J Mol Sci Article In this study, we confirmed that thrombin significantly increases the production of COX-2 and PGE(2) in human tracheal smooth muscle cells (HTSMCs), leading to inflammation in the airways and lungs. These molecules are well-known contributors to various inflammatory diseases. Here, we investigated in detail the involved signaling pathways using specific inhibitors and small interfering RNAs (siRNAs). Our results demonstrated that inhibitors targeting proteins such as protein kinase C (PKC)δ, proline-rich tyrosine kinase 2 (Pyk2), c-Src, epidermal growth factor receptor (EGFR), phosphatidylinositol 3-kinase (PI3K), or activator protein-1 (AP-1) effectively reduced thrombin-induced COX-2 and PGE(2) production. Additionally, transfection with siRNAs against PKCδ, Pyk2, c-Src, EGFR, protein kinase B (Akt), or c-Jun mitigated these responses. Furthermore, our observations revealed that thrombin stimulated the phosphorylation of key components of the signaling cascade, including PKCδ, Pyk2, c-Src, EGFR, Akt, and c-Jun. Thrombin activated COX-2 promoter activity through AP-1 activation, a process that was disrupted by a point-mutated AP-1 site within the COX-2 promoter. Finally, resveratrol (one of the most researched natural polyphenols) was found to effectively inhibit thrombin-induced COX-2 expression and PGE(2) release in HTSMCs through blocking the activation of Pyk2, c-Src, EGFR, Akt, and c-Jun. In summary, our findings demonstrate that thrombin-induced COX-2 and PGE(2) generation involves a PKCδ/Pyk2/c-Src/EGFR/PI3K/Akt-dependent AP-1 activation pathway. This study also suggests the potential use of resveratrol as an intervention for managing airway inflammation. MDPI 2023-10-13 /pmc/articles/PMC10606820/ /pubmed/37894811 http://dx.doi.org/10.3390/ijms242015130 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Yang, Chien-Chung
Lee, I-Ta
Lin, Yan-Jyun
Wu, Wen-Bin
Hsiao, Li-Der
Yang, Chuen-Mao
Thrombin-Induced COX-2 Expression and PGE(2) Synthesis in Human Tracheal Smooth Muscle Cells: Role of PKCδ/Pyk2-Dependent AP-1 Pathway Modulation
title Thrombin-Induced COX-2 Expression and PGE(2) Synthesis in Human Tracheal Smooth Muscle Cells: Role of PKCδ/Pyk2-Dependent AP-1 Pathway Modulation
title_full Thrombin-Induced COX-2 Expression and PGE(2) Synthesis in Human Tracheal Smooth Muscle Cells: Role of PKCδ/Pyk2-Dependent AP-1 Pathway Modulation
title_fullStr Thrombin-Induced COX-2 Expression and PGE(2) Synthesis in Human Tracheal Smooth Muscle Cells: Role of PKCδ/Pyk2-Dependent AP-1 Pathway Modulation
title_full_unstemmed Thrombin-Induced COX-2 Expression and PGE(2) Synthesis in Human Tracheal Smooth Muscle Cells: Role of PKCδ/Pyk2-Dependent AP-1 Pathway Modulation
title_short Thrombin-Induced COX-2 Expression and PGE(2) Synthesis in Human Tracheal Smooth Muscle Cells: Role of PKCδ/Pyk2-Dependent AP-1 Pathway Modulation
title_sort thrombin-induced cox-2 expression and pge(2) synthesis in human tracheal smooth muscle cells: role of pkcδ/pyk2-dependent ap-1 pathway modulation
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10606820/
https://www.ncbi.nlm.nih.gov/pubmed/37894811
http://dx.doi.org/10.3390/ijms242015130
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