G-Protein-coupled receptor agonists differentially regulate basal or tumor necrosis factor-α-stimulated activation of interleukin-6 and RANTES in human airway smooth muscle cells

Chien Da Huang, Alaina J. Ammit, Omar Tliba, Han Pin Kuo, Raymond B. Penn, Reynold A. Panettieri, Yassine Amrani

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Using thapsigargin (Tg), an agent that mobilizes calcium by directly emptying intracellular stores, we previously showed that intracellular calcium may play an important role in the regulation of intercellular adhesion molecule (ICAM)-1 gene expression induced by cytokines in human airway smooth muscle (ASM) cells. In the present study, we extended this previous observation by comparing the effect of Tg and other calcium-mobilizing G-protein-coupled receptor (GPCR) agonists on the expression of different pro-inflammatory genes in response to tumor necrosis factor (TNF)-α in ASM cells. We found that in resting cells, Tg (10-100 nM) or the bradykinin (BK) (1-10 μM) and thrombin (Thr) (1 U/ml) stimulated interleukin (IL)-6 secretion but had no effect on regulated on activation, normal T cells expressed and secreted (RANTES) levels. More importantly, such calcium-mobilizing agents significantly enhanced TNF-α-induced IL-6 secretion while RANTES secretion was abrogated. The use of luciferase-tagged IL-6 and RANTES promoter constructs demonstrated similar effects of Tg on IL-6 and RANTES genes in basal and TNF-α-stimulated conditions. The cyclic adenosine monophosphate (cAMP)-dependent pathway plays a minor role in this differential regulation of IL-6 and RANTES genes expression. 2-Aminoethoxydiphenyl borate (APB), a blocker of store-operated calcium channels (SOCs), and bisindolylmaleimide I (Bis I), a broad-spectrum protein kinase C (PKC) inhibitor, inhibited the basal and synergic effects of IL-6 secretion in response to calcium-mobilizing agents and TNF-α, but did not prevent the abrogated effect of RANTES secretion. We also found that Go-6976, a selective calcium-dependent PKC isozyme inhibitor, did not inhibit IL-6 secretion in response to GPCR agonist and TNF-α; whereas Rottlerlin, a PKC-δ inhibitor, inhibited both Thr- and TNF-α-induced expression of IL-6, while BK-induced IL-6 secretion was not affected. Interestingly, TNF-α-induced interferon regulatory factor (IRF)-1 activation was significantly inhibited by all calcium-mobilizing agents, BK, Thr and Tg. These results show that calcium-mobilizing GPCR agonists functionally interact with TNF-α to differentially regulate pro-inflammatory genes expression in human ASM cells, possibly by involving Tg-sensitive intracellular calcium stores, SOC and PKC.

Original languageEnglish
Pages (from-to)763-776
Number of pages14
JournalJournal of Biomedical Science
Volume12
Issue number5
DOIs
Publication statusPublished - Oct 1 2005
Externally publishedYes

Fingerprint

T-cells
G-Protein-Coupled Receptors
Smooth Muscle Myocytes
Muscle
Interleukin-6
Thapsigargin
Tumor Necrosis Factor-alpha
Chemical activation
Cells
T-Lymphocytes
Calcium
Protein Kinase C
Bradykinin
Gene expression
Thrombin
Protein C Inhibitor
Calcium Channels
Protein Kinase Inhibitors
Gene Expression
Genes

Keywords

  • Airway smooth muscle
  • Calcium
  • Differential regulation
  • GPCR agonists
  • IL-6 RANTES
  • PKC
  • Thapsigargin
  • TNF-α

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

G-Protein-coupled receptor agonists differentially regulate basal or tumor necrosis factor-α-stimulated activation of interleukin-6 and RANTES in human airway smooth muscle cells. / Huang, Chien Da; Ammit, Alaina J.; Tliba, Omar; Kuo, Han Pin; Penn, Raymond B.; Panettieri, Reynold A.; Amrani, Yassine.

In: Journal of Biomedical Science, Vol. 12, No. 5, 01.10.2005, p. 763-776.

Research output: Contribution to journalArticle

Huang, Chien Da ; Ammit, Alaina J. ; Tliba, Omar ; Kuo, Han Pin ; Penn, Raymond B. ; Panettieri, Reynold A. ; Amrani, Yassine. / G-Protein-coupled receptor agonists differentially regulate basal or tumor necrosis factor-α-stimulated activation of interleukin-6 and RANTES in human airway smooth muscle cells. In: Journal of Biomedical Science. 2005 ; Vol. 12, No. 5. pp. 763-776.
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T1 - G-Protein-coupled receptor agonists differentially regulate basal or tumor necrosis factor-α-stimulated activation of interleukin-6 and RANTES in human airway smooth muscle cells

AU - Huang, Chien Da

AU - Ammit, Alaina J.

AU - Tliba, Omar

AU - Kuo, Han Pin

AU - Penn, Raymond B.

AU - Panettieri, Reynold A.

AU - Amrani, Yassine

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N2 - Using thapsigargin (Tg), an agent that mobilizes calcium by directly emptying intracellular stores, we previously showed that intracellular calcium may play an important role in the regulation of intercellular adhesion molecule (ICAM)-1 gene expression induced by cytokines in human airway smooth muscle (ASM) cells. In the present study, we extended this previous observation by comparing the effect of Tg and other calcium-mobilizing G-protein-coupled receptor (GPCR) agonists on the expression of different pro-inflammatory genes in response to tumor necrosis factor (TNF)-α in ASM cells. We found that in resting cells, Tg (10-100 nM) or the bradykinin (BK) (1-10 μM) and thrombin (Thr) (1 U/ml) stimulated interleukin (IL)-6 secretion but had no effect on regulated on activation, normal T cells expressed and secreted (RANTES) levels. More importantly, such calcium-mobilizing agents significantly enhanced TNF-α-induced IL-6 secretion while RANTES secretion was abrogated. The use of luciferase-tagged IL-6 and RANTES promoter constructs demonstrated similar effects of Tg on IL-6 and RANTES genes in basal and TNF-α-stimulated conditions. The cyclic adenosine monophosphate (cAMP)-dependent pathway plays a minor role in this differential regulation of IL-6 and RANTES genes expression. 2-Aminoethoxydiphenyl borate (APB), a blocker of store-operated calcium channels (SOCs), and bisindolylmaleimide I (Bis I), a broad-spectrum protein kinase C (PKC) inhibitor, inhibited the basal and synergic effects of IL-6 secretion in response to calcium-mobilizing agents and TNF-α, but did not prevent the abrogated effect of RANTES secretion. We also found that Go-6976, a selective calcium-dependent PKC isozyme inhibitor, did not inhibit IL-6 secretion in response to GPCR agonist and TNF-α; whereas Rottlerlin, a PKC-δ inhibitor, inhibited both Thr- and TNF-α-induced expression of IL-6, while BK-induced IL-6 secretion was not affected. Interestingly, TNF-α-induced interferon regulatory factor (IRF)-1 activation was significantly inhibited by all calcium-mobilizing agents, BK, Thr and Tg. These results show that calcium-mobilizing GPCR agonists functionally interact with TNF-α to differentially regulate pro-inflammatory genes expression in human ASM cells, possibly by involving Tg-sensitive intracellular calcium stores, SOC and PKC.

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KW - Airway smooth muscle

KW - Calcium

KW - Differential regulation

KW - GPCR agonists

KW - IL-6 RANTES

KW - PKC

KW - Thapsigargin

KW - TNF-α

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