Lipopolysaccharide enhances bradykinin-induced signal transduction via activation of Ras/Raf/MEK/MAPK in canine tracheal smooth muscle cells

Shue Fen Luo, Chuan Chwan Wang, Chi Tso Chiu, Chin Sung Chien, Li Der Hsiao, Chien-Huang Lin, Chuen Mao Yang

Research output: Contribution to journalArticle

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Abstract

1. Bacterial lipopolysaccharide (LPS) was found to induce inflammatory responses and to enhance bronchial hyperreactivity to several contractile agonists. However, the implication of LPS in the pathogenesis of bronchial hyperreactivity was not completely understood. Therefore, in this study, we investigated the effect of LPS on mitogen-activated protein kinase (MAPK) activation associated with potentiation of bradykinin (BK)-induced inositol phosphates (IPs) accumulation and Ca 2+ mobilization in canine cultured tracheal smooth muscle cells (TSMCs). 2. LPS stimulated phosphorylation of p42/p44 MAPK in a time- and concentration-dependent manner using a Western blot analysis against a specific phosphorylated form of MAPK antibody. Maximal stimulation of the p42 and p44 MAPK isoforms occurred after 7 min-incubation and the maximal effect was achieved with 100 μg ml -1 LPS. 3. Pretreatment of TSMCs with LPS potentiated BK-induced IPs accumulation and Ca 2+ mobilization. However, there was no effect on the IPs response induced by endothelin-1, 5-hydroxytryptamine, and carbachol. In addition, pretreatment with PDGF-BB enhanced BK-induced IPs response. 4. These enhancements by LPS and PDGF-BB might be due to an increase in BK B 2 receptor density (B(max)) in TSMCs, characterized by competitive inhibition of [ 3H]-BK binding using B 1 and B 2 receptor-selective reagents. 5. The enhancing effects of LPS and PDGF-BB were attenuated by PD98059, an inhibitor of MAPK kinase (MEK), suggesting that the effect of LPS may share a common signalling pathway with PDGF-BB in TSMCs. 6. Furthermore, overexpression of dominant negative mutants, H-Ras-15A and Raf-N4, significantly suppressed p42/p44 MAPK activation induced by LPS and PDGF-BB, indicating that Ras and Raf may be required for activation of these kinases. 7 These results suggest that the augmentation of BK-induced responses produced by LPS might be, at least in part, mediated through activation of Ras/Raf/MEK/MAPK pathway in TSMCs.

Original languageEnglish
Pages (from-to)1799-1808
Number of pages10
JournalBritish Journal of Pharmacology
Volume130
Issue number8
Publication statusPublished - 2000

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Mitogen-Activated Protein Kinase Kinases
Bradykinin
Mitogen-Activated Protein Kinases
Smooth Muscle Myocytes
Lipopolysaccharides
Canidae
Signal Transduction
Inositol Phosphates
Mitogen-Activated Protein Kinase 1
Bronchial Hyperreactivity
Carbachol
Endothelin-1
Serotonin
Protein Isoforms
Phosphotransferases
Western Blotting
Phosphorylation
platelet-derived growth factor BB

Keywords

  • Bradykinin
  • Ca
  • Inositol phosphates
  • Lipopolysaccharide
  • MAPK
  • MEK

ASJC Scopus subject areas

  • Pharmacology

Cite this

Lipopolysaccharide enhances bradykinin-induced signal transduction via activation of Ras/Raf/MEK/MAPK in canine tracheal smooth muscle cells. / Luo, Shue Fen; Wang, Chuan Chwan; Chiu, Chi Tso; Chien, Chin Sung; Hsiao, Li Der; Lin, Chien-Huang; Yang, Chuen Mao.

In: British Journal of Pharmacology, Vol. 130, No. 8, 2000, p. 1799-1808.

Research output: Contribution to journalArticle

Luo, Shue Fen ; Wang, Chuan Chwan ; Chiu, Chi Tso ; Chien, Chin Sung ; Hsiao, Li Der ; Lin, Chien-Huang ; Yang, Chuen Mao. / Lipopolysaccharide enhances bradykinin-induced signal transduction via activation of Ras/Raf/MEK/MAPK in canine tracheal smooth muscle cells. In: British Journal of Pharmacology. 2000 ; Vol. 130, No. 8. pp. 1799-1808.
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T1 - Lipopolysaccharide enhances bradykinin-induced signal transduction via activation of Ras/Raf/MEK/MAPK in canine tracheal smooth muscle cells

AU - Luo, Shue Fen

AU - Wang, Chuan Chwan

AU - Chiu, Chi Tso

AU - Chien, Chin Sung

AU - Hsiao, Li Der

AU - Lin, Chien-Huang

AU - Yang, Chuen Mao

PY - 2000

Y1 - 2000

N2 - 1. Bacterial lipopolysaccharide (LPS) was found to induce inflammatory responses and to enhance bronchial hyperreactivity to several contractile agonists. However, the implication of LPS in the pathogenesis of bronchial hyperreactivity was not completely understood. Therefore, in this study, we investigated the effect of LPS on mitogen-activated protein kinase (MAPK) activation associated with potentiation of bradykinin (BK)-induced inositol phosphates (IPs) accumulation and Ca 2+ mobilization in canine cultured tracheal smooth muscle cells (TSMCs). 2. LPS stimulated phosphorylation of p42/p44 MAPK in a time- and concentration-dependent manner using a Western blot analysis against a specific phosphorylated form of MAPK antibody. Maximal stimulation of the p42 and p44 MAPK isoforms occurred after 7 min-incubation and the maximal effect was achieved with 100 μg ml -1 LPS. 3. Pretreatment of TSMCs with LPS potentiated BK-induced IPs accumulation and Ca 2+ mobilization. However, there was no effect on the IPs response induced by endothelin-1, 5-hydroxytryptamine, and carbachol. In addition, pretreatment with PDGF-BB enhanced BK-induced IPs response. 4. These enhancements by LPS and PDGF-BB might be due to an increase in BK B 2 receptor density (B(max)) in TSMCs, characterized by competitive inhibition of [ 3H]-BK binding using B 1 and B 2 receptor-selective reagents. 5. The enhancing effects of LPS and PDGF-BB were attenuated by PD98059, an inhibitor of MAPK kinase (MEK), suggesting that the effect of LPS may share a common signalling pathway with PDGF-BB in TSMCs. 6. Furthermore, overexpression of dominant negative mutants, H-Ras-15A and Raf-N4, significantly suppressed p42/p44 MAPK activation induced by LPS and PDGF-BB, indicating that Ras and Raf may be required for activation of these kinases. 7 These results suggest that the augmentation of BK-induced responses produced by LPS might be, at least in part, mediated through activation of Ras/Raf/MEK/MAPK pathway in TSMCs.

AB - 1. Bacterial lipopolysaccharide (LPS) was found to induce inflammatory responses and to enhance bronchial hyperreactivity to several contractile agonists. However, the implication of LPS in the pathogenesis of bronchial hyperreactivity was not completely understood. Therefore, in this study, we investigated the effect of LPS on mitogen-activated protein kinase (MAPK) activation associated with potentiation of bradykinin (BK)-induced inositol phosphates (IPs) accumulation and Ca 2+ mobilization in canine cultured tracheal smooth muscle cells (TSMCs). 2. LPS stimulated phosphorylation of p42/p44 MAPK in a time- and concentration-dependent manner using a Western blot analysis against a specific phosphorylated form of MAPK antibody. Maximal stimulation of the p42 and p44 MAPK isoforms occurred after 7 min-incubation and the maximal effect was achieved with 100 μg ml -1 LPS. 3. Pretreatment of TSMCs with LPS potentiated BK-induced IPs accumulation and Ca 2+ mobilization. However, there was no effect on the IPs response induced by endothelin-1, 5-hydroxytryptamine, and carbachol. In addition, pretreatment with PDGF-BB enhanced BK-induced IPs response. 4. These enhancements by LPS and PDGF-BB might be due to an increase in BK B 2 receptor density (B(max)) in TSMCs, characterized by competitive inhibition of [ 3H]-BK binding using B 1 and B 2 receptor-selective reagents. 5. The enhancing effects of LPS and PDGF-BB were attenuated by PD98059, an inhibitor of MAPK kinase (MEK), suggesting that the effect of LPS may share a common signalling pathway with PDGF-BB in TSMCs. 6. Furthermore, overexpression of dominant negative mutants, H-Ras-15A and Raf-N4, significantly suppressed p42/p44 MAPK activation induced by LPS and PDGF-BB, indicating that Ras and Raf may be required for activation of these kinases. 7 These results suggest that the augmentation of BK-induced responses produced by LPS might be, at least in part, mediated through activation of Ras/Raf/MEK/MAPK pathway in TSMCs.

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KW - Ca

KW - Inositol phosphates

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