VIP inhibits basal and histamine-stimulated proliferation of human airway smooth muscle cells

1995 ◽  
Vol 268 (6) ◽  
pp. L1047-L1051 ◽  
Author(s):  
K. Maruno ◽  
A. Absood ◽  
S. I. Said
Keyword(s):  
Smooth Muscle ◽  
Airway Smooth Muscle ◽  
Thymidine Incorporation ◽  
Inhibitory Effect ◽  
Airway Smooth Muscle Cells ◽  
Dependent Manner ◽  
Cyclic Monophosphate ◽  
Cell Counts ◽  
Dependent Protein ◽  
Camp Levels

Airway smooth muscle (ASM) cell proliferation contributes to increased airway resistance in bronchial asthma. We have examined the modulation of ASM proliferation by vasoactive intestinal peptide (VIP), a cotransmitter of airway relaxation. Human ASM cells were grown in culture as a monolayer. VIP (1.0 nM-1.0 microM) inhibited proliferation in a dose-dependent manner by up to 82% on day 2, but the related peptide glucagon had no effect. Histamine (100 nM-100 microM) increased cell counts by 66%, but in the presence of VIP, cell counts and [3H]thymidine incorporation were reduced by up to 55%. Adenosine 3',5'-cyclic monophosphate (cAMP)-promoting agents, including 3-isobutyl-1-methylxanthine, forskolin, and 8-bromo-adenosine 3',5'-cyclic monophosphate, alone and especially combined with VIP, reduced cell counts and [3H]thymidine incorporation, in correlation with cAMP levels. KT-5720 (1.0 nM-1.0 microM), a selective inhibitor of cAMP-dependent protein kinase A (PKA), abolished the inhibitory effect of VIP. The results show that VIP selectively and potently inhibits human ASM cell growth and multiplication, and nullifies the mitogenic effect of histamine, by a PKA-mediated mechanism. A deficiency of VIP may lead to ASM hyperplasia due to unopposed stimulation by endogenous mitogens.

Effects of cAMP on serotonin evoked calcium transients in cultured rat airway smooth muscle cells

1997 ◽  
Vol 272 (5) ◽  
pp. L865-L871 ◽  
Author(s):  
B. Tolloczko ◽  
Y. L. Jia ◽  
J. G. Martin
Keyword(s):  
Smooth Muscle ◽  
Protein Kinase ◽  
Airway Smooth Muscle ◽  
Airway Smooth Muscle Cells ◽  
Intracellular Camp ◽  
Dependent Protein Kinase ◽  
Cyclic Monophosphate ◽  
Dependent Protein ◽  
High Concentrations

Agents increasing intracellular adenosine 3',5'-cyclic monophosphate (cAMP) cause relaxation of airway smooth muscle. However, the mechanisms of their action are not fully understood. We investigated the role of cAMP in the modulation of intracellular Ca2+ concentration ([Ca2+]i) transients evoked by serotonin (5-HT) in cultured rat tracheal smooth muscle (TSM) cells. Forskolin (10(-7) M) caused a significant elevation of intracellular cAMP and a 60% relaxation of tracheal rings contracted with 5-HT but did not affect [Ca2+]i in TSM cells. Forskolin (10(-5) M) completely relaxed tracheal rings and significantly decreased [Ca2+]i during the sustained phase of the 5-HT response. Forskolin-induced relaxation was attenuated by the cAMP-dependent protein kinase A (PKA) inhibitor Rp diastereomer of cAMP (Rp-cAMPS; 10(-4) M) and by the guanosine 3',5'-cyclic monophosphate (cGMP)-dependent protein kinase (PKG) inhibitor [Rp isomer of 8-(4-chlorophenylthio)-guanosine 3',5'-cyclic monophosphorothioate, 10(-4) M]. The effects of forskolin on [Ca2+]i were not altered by the PKA inhibitor but were abolished by the PKG inhibitor and thapsigargin. These results indicate that, in rat TSM, the relaxant effects of high concentrations of cAMP may be mediated, at least in part, by facilitating the sequestration of Ca2+ into intracellular stores by a mechanism involving PKG.

Role of phospholipase C and tyrosine kinase systems in growth response of human airway smooth muscle cells

1996 ◽  
Vol 270 (5) ◽  
pp. L795-L802 ◽  
Author(s):  
S. De ◽  
E. T. Zelazny ◽  
J. F. Souhrada ◽  
M. Souhrada
Keyword(s):  
Smooth Muscle ◽  
Tyrosine Kinase ◽  
Cell Count ◽  
Phospholipase C ◽  
Airway Smooth Muscle ◽  
Thymidine Incorporation ◽  
Airway Smooth Muscle Cells ◽  
Dependent Manner ◽  
Human Airway Smooth Muscle ◽  
Human Airway

The primary culture of confluent human airway smooth muscle (ASM) cells were exposed up to 5 days to human recombinant interleukin (IL)-1 beta in the presence of indomethacin and 1% fetal bovine serum. The proliferation was assessed by a [3H]thymidine incorporation and direct cell count. We found that IL-1 beta significantly increased thymidine incorporation into and cell count of ASM cells in a concentration-dependent manner. Pretreatment of cells with specific polyclonal antibodies against platelet-derived growth factor (PDGF-BB homodimer) completely inhibited the IL-1 beta-induced increase in thymidine incorporation. The PDGF-BB, at the concentrations of 1.5 and 2.5 ng/ml, stimulated the proliferation of ASM cells. The proliferation action of IL-1 beta was potentiated when PDGF-BB was added into the medium in combination with IL-1 beta. Pretreatment of cells with genistein (0.37 microM), a specific tyrosine kinase inhibitor, attenuated the proliferative effect of IL-1 beta and PDGF-BB. To clarify whether these growth stimuli (IL-1 beta and PDGF-BB) activated phospholipase C (PLC), we examined the formation of phosphatidylinositols. We observed that both agents significantly increased phosphoinositide turnover. In contrast, genistein pretreatment (0.37 microM) prevented formation of inositol 1,4,5-trisphosphate (IP3), as induced by IL-1 beta and/or PDGF-BB. This study demonstrates that both IL-1 beta and PDGF-BB could induce proliferation of ASM cells through the activation of tyrosine kinase and PLC, which in turn stimulate the formation of IP3, a second messenger molecule.

Muscarinic inhibition of adenylyl cyclase regulates intracellular calcium in single airway smooth muscle cells

1996 ◽  
Vol 270 (2) ◽  
pp. L208-L214 ◽  
Author(s):  
J. M. Madison ◽  
H. Yamaguchi
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Intracellular Calcium ◽  
Adenylyl Cyclase ◽  
Airway Smooth Muscle ◽  
Muscle Cells ◽  
Inhibitory Effect ◽  
Airway Smooth Muscle Cells ◽  
Muscarinic Agonists ◽  
Cyclic Monophosphate

To determine whether muscarinic agonists attenuated isoproterenol-stimulated decreases in intracellular calcium concentration ([Ca2+]i), changes in [Ca2+]i were measured in single airway smooth muscle cells using ratiometric analysis of fura 2 fluorescence. Isoproterenol (10(-5) M) and 8-bromoadenosine 3',5'-cyclic monophosphate (8-BrcAMP) decreased [Ca2+]i by 24 +/- 3% (P < 0.05, n= 28) and 17 +/- 1% (P < 0.05, n = 6), respectively. The decreased [Ca2+]i in response to isoproterenol was inhibited by propranolol (10(-6) M) and Rp-adenosine 3',5'-cyclic monophosphothioate (Rp-cAMPS) (10-40 microM). In subsequent experiments assessing the effects of muscarinic agonists, isoproterenol did not decrease [Ca2+]i in the presence of carbachol (5 x 10(-8) M) (6 +/- 8% increase; NS, n = 8). To determine the mechanism underlying this inhibitory effect of carbachol, cells were loaded with 4,5-dimethoxy-2-nitrobenzyl adenosine-3',5'-cyclic monophosphate (caged cAMP). For cells loaded with 20 microM caged cAMP, photolysis of caged cAMP decreased basal [Ca2+]i by 28 +/- 3% (P < 0.05, n = 12). In the presence of carbachol (5 x 10(-8) M), photolysis of caged cAMP still induced a 27 +/- 4% decrease in [Ca2+]i (P < 0.05, n = 12). We concluded that a low concentration of carbachol did attenuate isoproterenol-stimulated decreases in [Ca2+]i. Because low concentrations of carbachol attenuated the decreases in [Ca2+]i stimulated by isoproterenol but not the comparable decreases stimulated by cAMP directly, we concluded that the inhibition of adenylyl cyclase activity by muscarinic agonists contributed to the regulation of [Ca2+]i in airway smooth muscle cells. The findings suggested that physiological levels of cholinergic stimulation inhibit adenylyl cyclase, thereby attenuating the effects that beta-adrenergic agonists have on [Ca2+]i.

IL-1 beta and IL-6 induce hyperplasia and hypertrophy of cultured guinea pig airway smooth muscle cells

1995 ◽  
Vol 78 (4) ◽  
pp. 1555-1563 ◽  
Author(s):  
S. De ◽  
E. T. Zelazny ◽  
J. F. Souhrada ◽  
M. Souhrada
Keyword(s):  
Smooth Muscle ◽  
Guinea Pig ◽  
Protein Content ◽  
Airway Smooth Muscle ◽  
Dna Content ◽  
Thymidine Incorporation ◽  
Interleukin 1 ◽  
Total Protein Content ◽  
Leucine Incorporation ◽  
Airway Smooth Muscle Cells

Guinea pig airway smooth muscle (ASM) cells were maintained in a primary tissue culture (passages 1–3). Cells were exposed to human recombinant interleukin-1 beta (IL-1 beta; 20–100 pg/ml) or interleukin-6 (IL-6; 1–4 ng/ml) in the presence of indomethacin (1 microgram/ml) for up to 5 days. Proliferation of ASM cells was assessed with two techniques, direct counting of cells with a hemacytometer and [3H]thymidine incorporation corrected for total protein content. Hypertrophy of ASM cells was assessed by [3H]leucine incorporation (evaluation of protein synthesis), determination of total DNA content, DNA content per cell, and protein content per cell. We observed that the exposure of ASM cells to human recombinant IL-1 beta or IL-6, in all studied concentrations, significantly increased the number of cells as well as [3H]thymidine incorporation into ASM cells. We also found that exposure of ASM to these two cytokines increased [3H]leucine incorporation into the ASM cells and increased protein content and DNA content per single cell. These changes were also concentration dependent. We conclude that the two proinflammatory cytokines, IL-1 beta and IL-6, which are present in asthmatic lungs, increased the proliferation of ASM cells (hyperplasia) as well as their overall size and size of their nuclei, as measured by biochemical markers. These findings are compatible with the presence of ASM hypertrophy.

Mechanisms of cyclic nucleotide-induced relaxation in canine tracheal smooth muscle

1995 ◽  
Vol 268 (3) ◽  
pp. L407-L413 ◽  
Author(s):  
I. McGrogan ◽  
S. Lu ◽  
S. Hipworth ◽  
L. Sormaz ◽  
R. Eng ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Cyclic Nucleotide ◽  
Cyclopiazonic Acid ◽  
Inhibitory Effect ◽  
Beta Agonist ◽  
Cyclic Monophosphate ◽  
Median Effective Concentration ◽  
Camp Levels ◽  
Ca2 Channels

The effects of exogeneous cyclopiazonic acid (CPA, 10 microM), a selective inhibitor of the sarcoplasmic reticulum (SR) Ca2+ adenosinetriphosphatase, on cyclic nucleotide-induced relaxations of canine airway smooth muscle were examined. Strips of tracheal muscle were precontracted with carbachol (50% median effective concentration, 0.1 microM) or with 60 mM KCl. The beta-agonist isoproterenol (ISO, 10 microM) relaxed the tissue by approximately 50%. The relaxation was reduced in the presence of CPA when L-type Ca2+ channels were available but not when these were blocked by 0.1 microM nifedipine. Forskolin (1.0 microM), an adenylate cyclase activator, was less effective at inhibiting the contraction than ISO, and addition of CPA did not block its inhibitory effect as effectively as when ISO was used. Radioimmunoassay indicated that both these agents raised adenosine 3',5'-cyclic monophosphate (cAMP) levels to the same degree. Very little relaxation of the precontracted smooth muscle was elicited by 3 mM 8-bromo-adenosine 3',5'-cyclic monophosphate (8-BrcAMP), and addition of CPA had no effect. Sodium nitroprusside (100 microM) and 8-bromo-guanosine 3',5'-cyclic monophosphate (10 mM) inhibited contraction to a greater degree than any agent that raised cAMP. These inhibitions were greatly reduced in the presence of CPA when L-type Ca2+ channels were available. We conclude that pumping of Ca2+ into SR plays a major role guanosine 3',5'-cyclic monophosphate-produced but not cAMP-induced relaxation; L-type Ca2+ channels must be available for the relaxant role of Ca2+ pumping into the SR to be expressed; and ISO-induced relaxation may not involve primarily elevation of the cAMP.(ABSTRACT TRUNCATED AT 250 WORDS)

scholarly journals Cytokines and Growth Factors Promote Airway Smooth Muscle Cell Proliferation

2012 ◽  
Vol 2012 ◽  
pp. 1-13 ◽  
Author(s):  
R. Stamatiou ◽  
E. Paraskeva ◽  
K. Gourgoulianis ◽  
P.-A. Molyvdas ◽  
A. Hatziefthimiou
Keyword(s):  
Smooth Muscle ◽  
Growth Factors ◽  
Smooth Muscle Cell ◽  
Airway Smooth Muscle ◽  
Muscle Cell ◽  
Thymidine Incorporation ◽  
Cell Number ◽  
Airway Smooth Muscle Cell ◽  
Specific Cell ◽  
Dependent Manner

Chronic airway diseases, such as asthma or chronic obstructive pulmonary disease, are characterized by the presence in the airways of inflammation factors, growth factors and cytokines, which promote airway wall remodelling. The aim of this study was to investigate the effect of cytokines and growth factors on airway smooth muscle cell (ASMC) proliferation, phenotype and responsiveness. Incubation of serum starved human bronchial ASMCs with TNF-α, TGF, bFGF, and PDGF, but not IL-1β, increased methyl-[3H]thymidine incorporation and cell number, mediated by the PI3K and MAPK signalling pathways. Regarding rabbit tracheal ASMC proliferation, TNF-α, IL-1β, TGF, and PDGF increased methyl-[3H]thymidine incorporation in a PI3K- and MAPK-dependent manner. bFGF increased both methyl-[3H]thymidine incorporation and cell number. Moreover, incubation with TGF, bFGF and PDGF appears to drive human ASMCs towards a synthetic phenotype, as shown by the reduction of the percentage of cells expressing SM-α actin. In addition, the responsiveness of epithelium-denuded rabbit tracheal strips to carbachol was not significantly altered after 3-day treatment with bFGF. In conclusion, all the tested cytokines and growth factors increased ASMC proliferation to a different degree, depending on the specific cell type, with bronchial ASMCs being more prone to proliferation than tracheal ASMCs.

Response of airway smooth muscle cells to TGF-beta 1: effects on growth and synthesis of glycosaminoglycans

1996 ◽  
Vol 271 (6) ◽  
pp. L910-L917 ◽  
Author(s):  
P. N. Black ◽  
P. G. Young ◽  
S. J. Skinner
Keyword(s):  
Smooth Muscle ◽  
Growth Factor ◽  
Smooth Muscle Cells ◽  
Cell Size ◽  
Airway Smooth Muscle ◽  
Thymidine Incorporation ◽  
Muscle Cells ◽  
Cell Number ◽  
Airway Smooth Muscle Cells ◽  
Tgf Beta

Transforming growth factor-beta (TGF-beta) is formed in the airways and may have a role in airway remodeling in asthma. We have studied the effects of TGF-beta on bovine airway smooth muscle cells (BASMC) in vitro. Thymidine incorporation by BASMC was inhibited after a 24-h incubation with TGF-beta 1. In contrast, thymidine incorporation by BASMC was stimulated (35.1 +/- 11.2%) after a 48-h incubation with 1 ng/ml TGF-beta 1. Cell number was also increased (25.9 +/- 7.6%) after a 72-h incubation with 3 ng/ml TGF-beta 1. TGF-beta 1 also increased cell size at 72 h, with a 24.3 +/- 6.2% increase in cell, diameter. Increases in BASMC size were accompanied by increased [3H]proline incorporation into cell protein. In cells from any individual animal, there was a strong inverse correlation (r = -0.97) between changes in cell number and cell size. In cells from some animals, the main effect of TGF-beta 1 was to promote an increase in cell number, whereas in others the predominant effect was cell hypertrophy. In contrast epidermal growth factor (EGF) led to an increase in thymidine incorporation and cell number in all preparations but did not increase cell size. TGF-beta 1 also promoted secretion of glycosaminoglycans into culture medium by BASMC with a preferential increase in hyaluronan secretion (4.5-fold) after 24 h. Latent TGF-beta (0.89 +/- 0.06 ng/ml) was also detected in conditioned medium from cultured BASMC, and TGF-beta 1 expression was demonstrated with RNA extracts from BASMC. Varying degrees of both smooth muscle cell hypertrophy and hyperplasia occur in asthma. These results obtained with airway smooth muscle cells indicate that TGF-beta could play a role in the structural changes seen in asthma.

scholarly journals The contribution of inositol 1,4,5-trisphosphate and ryanodine receptors to agonist-induced Ca2+ signaling of airway smooth muscle cells

2009 ◽  
Vol 297 (2) ◽  
pp. L347-L361 ◽  
Author(s):  
Yan Bai ◽  
Martin Edelmann ◽  
Michael J. Sanderson
Keyword(s):  
Wave Propagation ◽  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Airway Smooth Muscle ◽  
Ryanodine Receptors ◽  
Muscle Cells ◽  
Airway Smooth Muscle Cells ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Inositol 1,4,5 Trisphosphate

The relative contribution of inositol 1,4,5-trisphosphate (IP3) receptors (IP3Rs) and ryanodine receptors (RyRs) to agonist-induced Ca2+ signaling in mouse airway smooth muscle cells (SMCs) was investigated in lung slices with phase-contrast or laser scanning microscopy. At room temperature (RT), methacholine (MCh) or 5-hydroxytryptamine (5-HT) induced Ca2+ oscillations and an associated contraction in small airway SMCs. The subsequent exposure to an IP3R antagonist, 2-aminoethoxydiphenyl borate (2-APB), inhibited the Ca2+ oscillations and induced airway relaxation in a concentration-dependent manner. 2-APB also inhibited Ca2+ waves generated by the photolytic release of IP3. However, the RyR antagonist ryanodine had no significant effect, at any concentration, on airway contraction or agonist- or IP3-induced Ca2+ oscillations or Ca2+ wave propagation. By contrast, a second RyR antagonist, tetracaine, relaxed agonist-contracted airways and inhibited agonist-induced Ca2+ oscillations in a concentration-dependent manner. However, tetracaine did not affect IP3-induced Ca2+ release or wave propagation nor the Ca2+ content of SMC Ca2+ stores as evaluated by Ca2+-release induced by caffeine. Conversely, both ryanodine and tetracaine completely blocked agonist-independent slow Ca2+ oscillations induced by KCl. The inhibitory effects of 2-APB and absence of an effect of ryanodine on MCh-induced airway contraction or Ca2+ oscillations of SMCs were also observed at 37°C. In Ca2+-permeable SMCs, tetracaine inhibited agonist-induced contraction without affecting intracellular Ca2+ levels indicating that relaxation also resulted from a reduction in Ca2+ sensitivity. These results indicate that agonist-induced Ca2+ oscillations in mouse small airway SMCs are primary mediated via IP3Rs and that tetracaine induces relaxation by both decreasing Ca2+ sensitivity and inhibiting agonist-induced Ca2+ oscillations via an IP3-dependent mechanism.

Role of STIM1/Orai1-mediated store-operated Ca2+ entry in airway smooth muscle cell proliferation

2011 ◽  
Vol 110 (5) ◽  
pp. 1256-1263 ◽  
Author(s):  
Jin-jing Zou ◽  
Ya-dong Gao ◽  
Shuang Geng ◽  
Jiong Yang
Keyword(s):  
Smooth Muscle ◽  
Mrna Expression ◽  
Airway Smooth Muscle ◽  
Inhibitory Effect ◽  
Airway Smooth Muscle Cell ◽  
Characteristic Change ◽  
Airway Smooth Muscle Cells ◽  
Channel Blockers ◽  
Asthma Patients ◽  
Underlying Mechanisms

Hyperplasia of airway smooth muscle cells (ASMCs) is a characteristic change of chronic asthma patients. However, the underlying mechanisms that trigger this process are not yet completely understood. Store-operated Ca2+ (SOC) entry (SOCE) occurs in response to the intracellular sarcoplasma reticulum (SR)/endoplasmic reticulum (ER) Ca2+ store depletion. SOCE plays an important role in regulating Ca2+ signaling and cellular responses of ASMCs. Stromal interaction molecule (STIM)1 has been proposed as an ER/SR Ca2+ sensor and translocates to the ER underneath the plasma membrane upon depletion of the ER Ca2+ store, where it interacts with Orai1, the molecular component of SOC channels, and brings about SOCE. STIM1 and Orai1 have been proved to mediate SOCE of ASMCs. In this study, we investigated whether STIM1/Orai1-mediated SOCE is involved in rat ASMC proliferation. We found that SOCE was upregulated during ASMC proliferation accompanied by a mild increase of STIM1 and a significant increase of Orai1 mRNA expression, whereas the proliferation of ASMCs was partially inhibited by the SOC channel blockers SKF-96365, NiCl2, and BTP-2. Suppressing the mRNA expression of STIM1 or Orai1 with specific short hairpin RNA resulted in the attenuation of SOCE and ASMC proliferation. Moreover, after knockdown of STIM1 or Orai1, the SOC channel blocker SKF-96365 had no inhibitory effect on the proliferation of ASMCs anymore. These results suggested that STIM1/Orai1-mediated SOCE is involved in ASMC proliferation.

TGF-β1 stimulates IL-8 release, COX-2 expression, and PGE2release in human airway smooth muscle cells

2000 ◽  
Vol 279 (1) ◽  
pp. L201-L207 ◽  
Author(s):  
Choong Yi Fong ◽  
Linhua Pang ◽  
Elaine Holland ◽  
Alan J. Knox
Keyword(s):  
Smooth Muscle ◽  
Airway Smooth Muscle ◽  
Transforming Growth Factor ◽  
Airway Smooth Muscle Cells ◽  
Dependent Manner ◽  
Human Airway Smooth Muscle ◽  
Human Airway ◽  
Cox Inhibitor ◽  
Cox 2 ◽  
Tgf Β1

We have recently shown that endogenous prostanoids are critical in bradykinin-stimulated interleukin (IL)-8 release from human airway smooth muscle (ASM) cells. In this study, we tested the ability of transforming growth factor (TGF)-β1 to stimulate IL-8 release, cyclooxygenase (COX)-2 expression and PGE2 generation in cultured human ASM cells and explored the role of COX products and COX-2 induction on IL-8 release. TGF-β1 stimulated IL-8 release, COX-2 induction, and PGE2 generation in a concentration- and time-dependent manner. Maximal IL-8 release was achieved with 10 ng/ml of TGF-β1 after 16 h of incubation, which was inhibited by the transcription inhibitor actinomycin D and the corticosteroid dexamethasone but was not affected by the nonselective COX inhibitor indomethacin and the selective COX-2 inhibitor NS-398 despite their inhibition on TGF-β1-induced PGE2 release. These results show for the first time that TGF-β1 stimulates IL-8 release, COX-2 induction, and PGE2 generation in human ASM cells and that PGE2 generation is not critical for TGF-β1-induced IL-8 release. These findings suggest that TGF-β1 may play an important role in the pathophysiology of asthma.

Sign in / Sign up

Export Citation Format

Share Document