Role of ERK MAP kinases in responses of cultured  human airway smooth muscle cells to IL-1β

We have previously reported that interleukin (IL)-1β causes β-adrenergic hyporesponsiveness in cultured human airway smooth muscle cells by increasing cyclooxygenase-2 (COX-2) expression and prostanoid formation. The purpose of this study was to determine whether extracellular signal-regulated kinases (ERKs) are involved in these events. Levels of phosphorylated ERK (p42 and p44) increased 8.3- and 13-fold, respectively, 15 min after treatment with IL-1β (20 ng/ml) alone. Pretreating cells with the mitogen-activated protein kinase kinase inhibitor PD-98059 or U-126 (2 h before IL-1β treatment) decreased ERK phosphorylation. IL-1β (20 ng/ml for 22 h) alone caused a marked induction of COX-2 and increased basal PGE2 release 28-fold ( P < 0.001). PD-98059 (100 μM) and U-126 (10 μM) each decreased COX-2 expression when administered before IL-1β treatment. In control cells, PD-98059 and U-126 had no effect on basal or arachidonic acid (AA; 10 μM)-stimulated PGE2 release, but both inhibitors caused a significant decrease in bradykinin (BK; 1 μM)-stimulated PGE2 release, consistent with a role for ERK in the activation of phospholipase A2 by BK. In IL-1β-treated cells, prior administration of PD-98059 caused 81, 92 and 40% decreases in basal and BK- and AA-stimulated PGE2 release, respectively ( P < 0.01), whereas administration of PD-98059 20 h after IL-1β resulted in only 38 and 43% decreases in basal and BK-stimulated PGE2release, respectively ( P < 0.02) and had no effect on AA-stimulated PGE2 release. IL-1β attenuated isoproterenol-induced decreases in human airway smooth muscle stiffness as measured by magnetic twisting cytometry, and PD-98059 or U-126 abolished this effect in a concentration-dependent manner. These results are consistent with the hypothesis that ERKs are involved early in the signal transduction pathway through which IL-1β induces PGE2 synthesis and β-adrenergic hyporesponsiveness and that ERKs act by inducing COX-2 and activating phospholipase A2.

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Functional TRPV4 channels are expressed in human airway smooth muscle cells

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.00393.2003 ◽

2004 ◽

Vol 287 (2) ◽

pp. L272-L278 ◽

Cited By ~ 124

Author(s):

Yanlin Jia ◽

Xin Wang ◽

LoriAnn Varty ◽

Charles A. Rizzo ◽

Richard Yang ◽

...

Keyword(s):

Smooth Muscle ◽

Smooth Muscle Cells ◽

Airway Smooth Muscle ◽

Transient Receptor Potential ◽

Muscle Cells ◽

Ruthenium Red ◽

Airway Smooth Muscle Cells ◽

Dependent Manner ◽

Human Airway Smooth Muscle ◽

Human Airway

Hypotonic stimulation induces airway constriction in normal and asthmatic airways. However, the osmolarity sensor in the airway has not been characterized. TRPV4 (also known as VR-OAC, VRL-2, TRP12, OTRPC4), an osmotic-sensitive cation channel in the transient receptor potential (TRP) channel family, was recently cloned. In the present study, we show that TRPV4 mRNA was expressed in cultured human airway smooth muscle cells as analyzed by RT-PCR. Hypotonic stimulation induced Ca2+influx in human airway smooth muscle cells in an osmolarity-dependent manner, consistent with the reported biological activity of TRPV4 in transfected cells. In cultured muscle cells, 4α-phorbol 12,13-didecanoate (4-αPDD), a TRPV4 ligand, increased intracellular Ca2+level only when Ca2+was present in the extracellular solution. The 4-αPDD-induced Ca2+response was inhibited by ruthenium red (1 μM), a known TRPV4 inhibitor, but not by capsazepine (1 μM), a TRPV1 antagonist, indicating that 4-αPDD-induced Ca2+response is mediated by TRPV4. Verapamil (10 μM), an L-type voltage-gated Ca2+channel inhibitor, had no effect on the 4-αPDD-induced Ca2+response, excluding the involvement of L-type Ca2+channels. Furthermore, hypotonic stimulation elicited smooth muscle contraction through a mechanism dependent on membrane Ca2+channels in both isolated human and guinea pig airways. Hypotonicity-induced airway contraction was not inhibited by the L-type Ca2+channel inhibitor nifedipine (1 μM) or by the TRPV1 inhibitor capsazepine (1 μM). We conclude that functional TRPV4 is expressed in human airway smooth muscle cells and may act as an osmolarity sensor in the airway.

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Faculty Opinions recommendation of Macrolides inhibit IL17-induced IL8 and 8-isoprostane release from human airway smooth muscle cells.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.718534594.793498785 ◽

2014 ◽

Author(s):

Charles Feldman

Keyword(s):

Smooth Muscle ◽

Smooth Muscle Cells ◽

Airway Smooth Muscle ◽

Muscle Cells ◽

Airway Smooth Muscle Cells ◽

Human Airway Smooth Muscle ◽

Human Airway

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TNFα and IFNγ Phosphorylate RNA Polymerase II To Synergistically Enhance CXCL-10 Release from Human Airway Smooth Muscle Cells.

10.1164/ajrccm-conference.2009.179.1_meetingabstracts.a6312 ◽

2009 ◽

Author(s):

DL Clarke ◽

S Jindarat ◽

D Proud ◽

L Corbett ◽

L Pang ◽

...

Keyword(s):

Smooth Muscle ◽

Rna Polymerase ◽

Smooth Muscle Cells ◽

Rna Polymerase Ii ◽

Airway Smooth Muscle ◽

Muscle Cells ◽

Airway Smooth Muscle Cells ◽

Human Airway Smooth Muscle ◽

Human Airway

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TNFα and IFNγ Synergistically Enhance Transcriptional Activation ofCXCL10in Human Airway Smooth Muscle Cells via STAT-1, NF-κB, and the Transcriptional Coactivator CREB-binding Protein

Journal of Biological Chemistry ◽

10.1074/jbc.m109.099952 ◽

2010 ◽

Vol 285 (38) ◽

pp. 29101-29110 ◽

Cited By ~ 57

Author(s):

Deborah L. Clarke ◽

Rachel L. Clifford ◽

Sarawut Jindarat ◽

David Proud ◽

Linhua Pang ◽

...

Keyword(s):

Smooth Muscle ◽

Smooth Muscle Cells ◽

Airway Smooth Muscle ◽

Transcriptional Activation ◽

Muscle Cells ◽

Airway Smooth Muscle Cells ◽

Creb Binding Protein ◽

Transcriptional Coactivator ◽

Human Airway Smooth Muscle ◽

Human Airway

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Urokinase potentiates PDGF-induced chemotaxis of human airway smooth muscle cells

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.00092.2002 ◽

2003 ◽

Vol 284 (6) ◽

pp. L1020-L1026 ◽

Cited By ~ 34

Author(s):

Stephen M. Carlin ◽

Michael Roth ◽

Judith L. Black

Keyword(s):

Smooth Muscle ◽

Smooth Muscle Cells ◽

Airway Smooth Muscle ◽

Kinase Inhibitor ◽

Airway Remodeling ◽

Muscle Cells ◽

Mek Inhibitor ◽

Airway Smooth Muscle Cells ◽

Human Airway Smooth Muscle ◽

Human Airway

We investigated the chemotactic action of PDGF and urokinase on human airway smooth muscle (HASM) cells in culture. Cells were put in collagen-coated transwells with 8-μm perforations, incubated for 4 h with test compounds, then fixed, stained, and counted as migrated nuclei by microscopy. Cells from all culture conditions showed some basal migration (migration in the absence of stimuli during the assay), but cells preincubated for 24 h in 10% FBS or 20 ng/ml PDGF showed higher basal migration than cells quiesced in 1% FBS. PDGFBB, PDGFAA, and PDGFABwere all chemotactic when added during the assay. PDGF chemotaxis was blocked by the phosphatidyl 3′-kinase inhibitor LY-294002, the MEK inhibitor U-0126, PGE2, formoterol, pertussis toxin, and the Rho kinase inhibitor Y-27632. Urokinase alone had no stimulatory effect on migration of quiescent cells but caused a dose-dependent potentiation of chemotaxis toward PDGF. Urokinase also potentiated the elevated basal migration of cells pretreated in 10% FBS or PDGF. This potentiating effect of urokinase appears to be novel. We conclude that PDGF and similar cytokines may be important factors in airway remodeling by redistribution of smooth muscle cells during inflammation and that urokinase may be important in potentiating the response.

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