Cargando…

Thrombin Induces COX-2 and PGE(2) Expression via PAR1/PKCalpha/MAPK-Dependent NF-kappaB Activation in Human Tracheal Smooth Muscle Cells

The inflammation of the airway and lung could be triggered by upregulation cyclooxygenase (COX)-2 and prostaglandin E(2) (PGE(2)) induced by various proinflammatory factors. COX-2 induction by thrombin has been shown to play a vital role in various inflammatory diseases. However, in human tracheal s...

Descripción completa

Detalles Bibliográficos
Autores principales: Yang, Chien-Chung, Hsiao, Li-Der, Shih, Ya-Fang, Hsu, Chih-Kai, Hu, Chia-Yu, Yang, Chuen-Mao
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Hindawi 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9042634/
https://www.ncbi.nlm.nih.gov/pubmed/35497094
http://dx.doi.org/10.1155/2022/4600029
_version_ 1784694702428127232
author Yang, Chien-Chung
Hsiao, Li-Der
Shih, Ya-Fang
Hsu, Chih-Kai
Hu, Chia-Yu
Yang, Chuen-Mao
author_facet Yang, Chien-Chung
Hsiao, Li-Der
Shih, Ya-Fang
Hsu, Chih-Kai
Hu, Chia-Yu
Yang, Chuen-Mao
author_sort Yang, Chien-Chung
collection PubMed
description The inflammation of the airway and lung could be triggered by upregulation cyclooxygenase (COX)-2 and prostaglandin E(2) (PGE(2)) induced by various proinflammatory factors. COX-2 induction by thrombin has been shown to play a vital role in various inflammatory diseases. However, in human tracheal smooth muscle cells (HTSMCs), how thrombin enhanced the levels of COX-2/PGE(2) is not completely characterized. Thus, in this study, the levels of COX-2 expression and PGE(2) synthesis induced by thrombin were determined by Western blot, promoter-reporter assay, real-time PCR, and ELISA kit. The various signaling components involved in the thrombin-mediated responses were differentiated by transfection with siRNAs and selective pharmacological inhibitors. The role of NF-κB was assessed by a chromatin immunoprecipitation (ChIP) assay, immunofluorescent staining, as well as Western blot. Our results verified that thrombin markedly triggered PGE(2) secretion via COX-2 upregulation which were diminished by the inhibitor of thrombin (PPACK), PAR1 (SCH79797), G(i/o) protein (GPA2), G(q) protein (GPA2A), PKCα (Gö6976), p38 MAPK (SB202190), JNK1/2 (SP600125), MEK1/2 (U0126), or NF-κB (helenalin) and transfection with siRNA of PAR1, G(q)α, G(i)α, PKCα, JNK2, p38, p42, or p65. Moreover, thrombin induced PAR1-dependent PKCα phosphorylation in HTSMCs. We also observed that thrombin induced p38 MAPK, JNK1/2, and p42/p44 MAPK activation through a PAR1/PKCα pathway. Thrombin promoted phosphorylation of NF-κB p65, leading to nuclear translocation and binding to the COX-2 promoter element to enhance promoter activity, which was reduced by Gö6976, SP600125, SB202190, or U0126. These findings supported that COX-2/PGE(2) expression triggered by thrombin was engaged in PAR1/G(q) or G(i/o)/PKCα/MAPK-dependent NF-κB activation in HTSMCs.
format Online
Article
Text
id pubmed-9042634
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher Hindawi
record_format MEDLINE/PubMed
spelling pubmed-90426342022-04-27 Thrombin Induces COX-2 and PGE(2) Expression via PAR1/PKCalpha/MAPK-Dependent NF-kappaB Activation in Human Tracheal Smooth Muscle Cells Yang, Chien-Chung Hsiao, Li-Der Shih, Ya-Fang Hsu, Chih-Kai Hu, Chia-Yu Yang, Chuen-Mao Mediators Inflamm Research Article The inflammation of the airway and lung could be triggered by upregulation cyclooxygenase (COX)-2 and prostaglandin E(2) (PGE(2)) induced by various proinflammatory factors. COX-2 induction by thrombin has been shown to play a vital role in various inflammatory diseases. However, in human tracheal smooth muscle cells (HTSMCs), how thrombin enhanced the levels of COX-2/PGE(2) is not completely characterized. Thus, in this study, the levels of COX-2 expression and PGE(2) synthesis induced by thrombin were determined by Western blot, promoter-reporter assay, real-time PCR, and ELISA kit. The various signaling components involved in the thrombin-mediated responses were differentiated by transfection with siRNAs and selective pharmacological inhibitors. The role of NF-κB was assessed by a chromatin immunoprecipitation (ChIP) assay, immunofluorescent staining, as well as Western blot. Our results verified that thrombin markedly triggered PGE(2) secretion via COX-2 upregulation which were diminished by the inhibitor of thrombin (PPACK), PAR1 (SCH79797), G(i/o) protein (GPA2), G(q) protein (GPA2A), PKCα (Gö6976), p38 MAPK (SB202190), JNK1/2 (SP600125), MEK1/2 (U0126), or NF-κB (helenalin) and transfection with siRNA of PAR1, G(q)α, G(i)α, PKCα, JNK2, p38, p42, or p65. Moreover, thrombin induced PAR1-dependent PKCα phosphorylation in HTSMCs. We also observed that thrombin induced p38 MAPK, JNK1/2, and p42/p44 MAPK activation through a PAR1/PKCα pathway. Thrombin promoted phosphorylation of NF-κB p65, leading to nuclear translocation and binding to the COX-2 promoter element to enhance promoter activity, which was reduced by Gö6976, SP600125, SB202190, or U0126. These findings supported that COX-2/PGE(2) expression triggered by thrombin was engaged in PAR1/G(q) or G(i/o)/PKCα/MAPK-dependent NF-κB activation in HTSMCs. Hindawi 2022-04-19 /pmc/articles/PMC9042634/ /pubmed/35497094 http://dx.doi.org/10.1155/2022/4600029 Text en Copyright © 2022 Chien-Chung Yang et al. https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Yang, Chien-Chung
Hsiao, Li-Der
Shih, Ya-Fang
Hsu, Chih-Kai
Hu, Chia-Yu
Yang, Chuen-Mao
Thrombin Induces COX-2 and PGE(2) Expression via PAR1/PKCalpha/MAPK-Dependent NF-kappaB Activation in Human Tracheal Smooth Muscle Cells
title Thrombin Induces COX-2 and PGE(2) Expression via PAR1/PKCalpha/MAPK-Dependent NF-kappaB Activation in Human Tracheal Smooth Muscle Cells
title_full Thrombin Induces COX-2 and PGE(2) Expression via PAR1/PKCalpha/MAPK-Dependent NF-kappaB Activation in Human Tracheal Smooth Muscle Cells
title_fullStr Thrombin Induces COX-2 and PGE(2) Expression via PAR1/PKCalpha/MAPK-Dependent NF-kappaB Activation in Human Tracheal Smooth Muscle Cells
title_full_unstemmed Thrombin Induces COX-2 and PGE(2) Expression via PAR1/PKCalpha/MAPK-Dependent NF-kappaB Activation in Human Tracheal Smooth Muscle Cells
title_short Thrombin Induces COX-2 and PGE(2) Expression via PAR1/PKCalpha/MAPK-Dependent NF-kappaB Activation in Human Tracheal Smooth Muscle Cells
title_sort thrombin induces cox-2 and pge(2) expression via par1/pkcalpha/mapk-dependent nf-kappab activation in human tracheal smooth muscle cells
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9042634/
https://www.ncbi.nlm.nih.gov/pubmed/35497094
http://dx.doi.org/10.1155/2022/4600029
work_keys_str_mv AT yangchienchung thrombininducescox2andpge2expressionviapar1pkcalphamapkdependentnfkappabactivationinhumantrachealsmoothmusclecells
AT hsiaolider thrombininducescox2andpge2expressionviapar1pkcalphamapkdependentnfkappabactivationinhumantrachealsmoothmusclecells
AT shihyafang thrombininducescox2andpge2expressionviapar1pkcalphamapkdependentnfkappabactivationinhumantrachealsmoothmusclecells
AT hsuchihkai thrombininducescox2andpge2expressionviapar1pkcalphamapkdependentnfkappabactivationinhumantrachealsmoothmusclecells
AT huchiayu thrombininducescox2andpge2expressionviapar1pkcalphamapkdependentnfkappabactivationinhumantrachealsmoothmusclecells
AT yangchuenmao thrombininducescox2andpge2expressionviapar1pkcalphamapkdependentnfkappabactivationinhumantrachealsmoothmusclecells