Inhibitors of Mitogen-activated Protein Kinases Differentially Regulate Eosinophil-activating Cytokine Release from Human Airway Smooth Muscle

Hallsworth, Matthew P.; Moir, Lyn M.; Lai, Dilys; Hirst, Stuart J John

doi:10.1164/ajrccm.164.4.2011004

Cited by 70 publications

(74 citation statements)

References 38 publications

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“…Similarly, the effect of SB-203580 on cytokine-induced mediator release has also been evaluated in human cultured ASMC. SB-203580 specifically inhibited cytokineinduced p38 kinase activity at concentrations up to 10 M and only partially inhibited JNK and ERK activity at 100 M (29). It is unlikely that inhibition of JNK activity contributed to the inhibitory action of SB-203580 on s-FKN release observed at the 20 M concentration used in this study.…”

Section: Discussionmentioning

confidence: 61%

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Fractalkine/CX₃CL1 production by human airway smooth muscle cells: induction by IFN-γ and TNF-α and regulation by TGF-β and corticosteroids

Sukkar

Issa

Xie

et al. 2004

American Journal of Physiology-Lung Cellular and Molecular Phys

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mokine synthesis by airway smooth muscle cells (ASMC) may be an important process underlying inflammatory cell recruitment in airway inflammatory diseases such as asthma and chronic obstructive pulmonary disease (COPD). Fractalkine (FKN) is a recently described CX 3C chemokine that has dual functions, serving as both a cell adhesion molecule and a chemoattractant for monocytes and T cells, expressing its unique receptor, CX3CR1. We investigated FKN expression by human ASMC in response to the proinflammatory cytokines IL-1␤, TNF-␣, and IFN-␥, the T helper 2-type cytokines IL-4, IL-10, and IL-13, and the fibrogenic cytokine transforming growth factor (TGF)-␤. Neither of these cytokines alone had any significant effect on ASMC FKN production. Combined stimulation with IFN-␥ and TNF-␣ induced FKN mRNA and protein expression in a time-and concentration-dependent manner. TGF-␤ had a significant inhibitory effect on cytokine-induced FKN mRNA and protein expression. Dexamethasone (10 Ϫ8 -10 Ϫ6 M) significantly upregulated cytokineinduced FKN mRNA and protein expression. Finally, we used selective inhibitors of the mitogen-activated protein kinases c-Jun NH2-terminal kinase (JNK) (SP-610025), p38 (SB-203580), and extracellular signal-regulated kinase (PD-98095) to investigate their role in FKN production. SP-610025 (25 M) and SB-203580 (20 M), but not PD-98095, significantly attenuated cytokine-induced FKN protein synthesis. IFN-␥-and TNF-␣-induced JNK phosphorylation remained unaltered in the presence of TGF-␤ but was inhibited by dexamethasone, indicating that JNK is not involved in TGF-␤-or dexamethasone-mediated regulation of FKN production. In summary, FKN production by human ASMC in vitro is regulated by inflammatory and anti-inflammatory factors. chemokine; airway inflammation; asthma; chronic obstructive pulmonary disease THERE IS NOW SUBSTANTIAL EVIDENCE suggesting that airway smooth muscle cells (ASMC), by virtue of their immunomodulatory functions, may be involved in regulating airway inflammatory responses in diseases such as asthma and chronic obstructive pulmonary disease (COPD). ASMC express adhesion molecules and synthesize many cytokines and chemokines such as granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-8, eotaxin, and RANTES. These mediators are critically implicated in the recruitment, survival, and activation of key effector cells, such as eosinophils, neutrophils, and T lymphocytes, in asthma and COPD (10).Fractalkine (FKN), also known as CX 3 C ligand 1 (CX 3 CL1), is a novel chemokine belonging to the CX 3 C chemokine family. FKN is a multidomain molecule expressed on the cell surface and consists of a transmembrane region and a heavily glycosylated mucin-like stalk that extends from the cell surface, holding the chemokine domain at its tip (5). FKN also exists as a soluble glycoprotein that is generated by proteolytic cleavage of the full-length molecule at a membrane-proximal site (27,32,55). The physiological relevance of membrane and soluble FKN is not known; however, it is cu...

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Section: Discussionmentioning

confidence: 61%

“…ASMC secretory responses of certain cytokines and chemokines are, in part, mediated by activation of the MAPK pathway (2,4,29,47). MAPK are a family of serine/threonine kinases, and at least three major groups that differ in their substrate specificity have been characterized: ERK, JNK, and p38 MAPK.…”

Section: Discussionmentioning

confidence: 99%

Fractalkine/CX₃CL1 production by human airway smooth muscle cells: induction by IFN-γ and TNF-α and regulation by TGF-β and corticosteroids

Sukkar

Issa

Xie

et al. 2004

American Journal of Physiology-Lung Cellular and Molecular Phys

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“…Davies et al (9) found that U-0126 reduced the in vitro kinase activity of p38␣ MAPK by ϳ25%. In contrast, studies in rat cardiomyocytes (45) and in human airway smooth muscle cells (17) have shown that U-0126 selectively inhibits p44/p42 MAPK activity without affecting p38 MAPK activity. To determine whether U-0126 inhibits the activity of p38 MAPK in response to insulin in adipocytes, we conducted an in vitro p38 MAPK assay.…”

Section: Impaired Phosphorylation Of P44/p42 Mapk By Pi3k Inhibitorsmentioning

confidence: 87%

MEK inhibitors impair insulin-stimulated glucose uptake in 3T3-L1 adipocytes

Harmon¹,

Paul²,

Patel³

2004

American Journal of Physiology-Endocrinology and Metabolism

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.-In 3T3-L1 adipocytes, insulin activates three major signaling cascades, the phosphoinositide 3-kinase (PI3K) pathway, the Cbl pathway, and the mitogen-activated protein kinase (MAPK) pathway. Although PI3K and Cbl mediate insulin-stimulated glucose uptake by promoting the translocation of the insulin-responsive glucose transporter (GLUT4) to the plasma membrane, the MAPK pathway does not have an established role in insulin-stimulated glucose uptake. We demonstrate in this report that PI3K inhibitors also inhibit the MAPK pathway. To investigate the role of the MAPK pathway separately from that of the PI3K pathway in insulin-stimulated glucose uptake, we used two specific inhibitors of MAPK kinase (MEK) activity, PD-98059 and U-0126, which reduced insulin-stimulated glucose uptake by ϳ33 and 50%, respectively. Neither MEK inhibitor affected the activation of Akt or PKC/, downstream signaling molecules in the PI3K pathway. Inhibition of MEK with U-0126 did not prevent GLUT4 from translocating to the plasma membrane, nor did it inhibit the subsequent docking and fusion of GLUT4-myc with the plasma membrane. MEK inhibitors affected glucose transport mediated by GLUT4 but not GLUT1. Importantly, the presence of MEK inhibitors only at the time of the transport assay markedly impaired both insulin-stimulated glucose uptake and MAPK signaling. Conversely, removal of MEK inhibitors before the transport assay restored glucose uptake and MAPK signaling. Collectively, our studies suggest a possible role for MEK in the activation of GLUT4. p44/p42 mitogen-activated protein kinase; glucose transporter 4; glucose transport; U-0126; PD-98059 THE BINDING OF INSULIN to its receptor triggers multiple signaling pathways that participate broadly in cellular growth and differentiation and in the metabolism of lipids and glucose. To maintain glucose homeostasis, insulin not only inhibits the production of glucose in the liver but also stimulates the transport of glucose into muscle and adipose tissue for subsequent use or storage. Autophosphorylation of the insulin receptor tyrosine kinase results in the recruitment and tyrosine phosphorylation of substrates and the activation of three major pathways, the phosphoinositide 3-kinase (PI3K) pathway, the mitogen-activated protein kinase (MAPK) pathway, and the Cbl pathway (see review in Ref.3). PI3K catalyzes the formation of phosphatidylinositol-3,4,5-trisphosphate, or PIP 3 , an allosteric activator of phosphoinositide-dependent kinase (PDK). Targets of PDK include Akt and the atypical PKC isoforms, which, when activated via phosphorylation, stimulate the translocation of the insulin-responsive glucose transporter (GLUT4)-containing vesicles to the plasma membrane. Phospholipase C-␥1 (PLC-␥1), another downstream target of PI3K in insulin signaling, also facilitates GLUT4 translocation (13,25). Although the relative contributions of PLC-␥1, Akt, and atypical PKCs to GLUT4 translocation remain controversial, studies have demonstrated the necessity of PI3K activity for insulin-stimulated...

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“…The release of several cytokines, GM-CSF, regulated on activation normal T cells expressed and secreted (RANTES), and eotaxin, is dependent on the activation of the MAPKs in a complex fashion (9). Hallsworth et al (9) showed that GM-CSF release was dependent on the activation of p42/p44 but was suppressed by the activation of p38, and more recently Oltmanns et al (16) demonstrated the involvement of JNK in the TNF-␣-and IL-1␤-induced expression of GM-CSF, IL-8, and RANTES. These few examples demonstrate the complexity of cell responses to activation of the MAPKs.…”

Section: Discussionmentioning

confidence: 99%

Mechanisms of serum potentiation of GM-CSF production by human airway smooth muscle cells

Lalor¹,

Truong²,

Henness³

et al. 2004

American Journal of Physiology-Lung Cellular and Molecular Phys

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Inflammation and vascular leakage are prevalent in asthma. This study aimed to elucidate the mechanisms involved in serum potentiation of cytokine-induced granulocyte macrophage colony stimulating factor (GM-CSF) production by human airway smooth muscle cells and to identify possible factors responsible. Serum-deprived cells at low density were stimulated with TNF-α and IL-1β for 24 h. Human AB serum (10%), inhibitors of RNA and protein synthesis or specific signaling molecules, or known smooth muscle mitogens were then added for 24 h. Culture supernatants were analyzed for GM-CSF levels, and cells were harvested to assess viability, cell cycle progression, GM-CSF-specific mRNA content, and p38 phosphorylation. Serum potentiated GM-CSF release when added before, together with (maximal), or after the cytokines. The potentiation involved both new GM-CSF-specific mRNA production and protein synthesis. The mitogens IGF, PDGF, and thrombin all potentiated GM-CSF release, and neutralizing antibodies for EGF, IGF, and PDGF reduced the serum potentiation. Inhibitor studies ruled as unlikely the involvement of p70S6kinase and the MAPK p42/p44, two signaling pathways implicated in proliferation, and the involvement of the MAPK JNK, while establishing roles for p38 MAPK and NF-κB in the potentiation of GM-CSF release. Detection of significant p38 phosphorylation in response to serum stimulation, through Western blotting, further demonstrated the involvement of p38. These studies have provided evidence to support p38 being targeted to interrupt the cycle of inflammation, vascular leakage and cytokine production in asthma.

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Inhibitors of Mitogen-activated Protein Kinases Differentially Regulate Eosinophil-activating Cytokine Release from Human Airway Smooth Muscle

Cited by 70 publications

References 38 publications

Fractalkine/CX₃CL1 production by human airway smooth muscle cells: induction by IFN-γ and TNF-α and regulation by TGF-β and corticosteroids

Fractalkine/CX₃CL1 production by human airway smooth muscle cells: induction by IFN-γ and TNF-α and regulation by TGF-β and corticosteroids

MEK inhibitors impair insulin-stimulated glucose uptake in 3T3-L1 adipocytes

Mechanisms of serum potentiation of GM-CSF production by human airway smooth muscle cells

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Inhibitors of Mitogen-activated Protein Kinases Differentially Regulate Eosinophil-activating Cytokine Release from Human Airway Smooth Muscle

Cited by 70 publications

References 38 publications

Fractalkine/CX3CL1 production by human airway smooth muscle cells: induction by IFN-γ and TNF-α and regulation by TGF-β and corticosteroids

Fractalkine/CX3CL1 production by human airway smooth muscle cells: induction by IFN-γ and TNF-α and regulation by TGF-β and corticosteroids

MEK inhibitors impair insulin-stimulated glucose uptake in 3T3-L1 adipocytes

Mechanisms of serum potentiation of GM-CSF production by human airway smooth muscle cells

Contact Info

Product

Resources

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Fractalkine/CX₃CL1 production by human airway smooth muscle cells: induction by IFN-γ and TNF-α and regulation by TGF-β and corticosteroids

Fractalkine/CX₃CL1 production by human airway smooth muscle cells: induction by IFN-γ and TNF-α and regulation by TGF-β and corticosteroids