Effects of neuropeptides on human lung fibroblast proliferation and chemotaxis

An increase in subepithelial mesenchymal cells and associated connective tissue is a feature of bronchial asthma. We determined whether neuropeptides could modulate fibroblast activity, particularly with respect to proliferation and chemotaxis. Human lung fibroblasts were cultured with neurokinin A (NKA), substance P (SP), vasoactive intestinal peptide (VIP), and calcitonin-gene-related peptide (CGRP). After 48 h, fibroblast proliferation was measured by a colorimetric assay based on the uptake and subsequent release of methylene blue. The chemotactic response to neuropeptides was determined with the use of a modified Boyden chamber. Both NKA and SP (10(-7)-10(-4) M) stimulated human lung fibroblast proliferation in HFL1 and IMR-90 fibroblasts. VIP and CGRP had no effect on fibroblast proliferation. NKA alone stimulated fibroblast chemotaxis maximally at 10(-10) M. Neutral endopeptidase (NEP) activity of 0.52 and 5.2 pmol/10(6) cells was assayed in IMR-90 and Hs68 fibroblasts, respectively. Phosphoramidon (5 x 10(-6)-10(-5) M), an NEP inhibitor, enhanced fibroblast proliferation in a dose-dependent manner. Thus neuropeptides have the potential to cause activation of mesenchymal cells, and neuropeptide release may contribute to the structural abnormalities observed in asthmatic airways.

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Glucocorticoid-induced contractility and F-actin content of human lung fibroblasts in three-dimensional culture

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.2000.278.1.l13 ◽

2000 ◽

Vol 278 (1) ◽

pp. L13-L18 ◽

Cited By ~ 5

Author(s):

Hiroyuki Miki ◽

Tadashi Mio ◽

Sonoko Nagai ◽

Yuma Hoshino ◽

Takeo Tsutsumi ◽

...

Keyword(s):

Wound Healing ◽

Human Lung ◽

Healing Process ◽

Lung Fibroblast ◽

Lung Fibroblasts ◽

Wound Healing Process ◽

Architectural Distortion ◽

Type I ◽

Dependent Manner ◽

Human Lung Fibroblast

Fibroblast contractility plays a useful role in the wound healing process but contributes to architectural distortion in the lungs. Glucocorticoids (GCs) have been reported to reduce dermal fibroblast contractility, which may result in delaying wound healing, but the effects on lung fibroblasts are unknown. In this study, we examined how human lung fibroblast contractility is altered in the presence of GCs. Lung fibroblast cell lines ( n = 5) were established from normal parts of surgically resected lung tissue. The effects of GCs on contractility were investigated with a type I collagen gel contraction assay. Filamentous actin (F-actin) content was detected by confocal microscopy and measured with a fluorescent phalloidin binding assay. GCs augmented fibroblast contraction in a concentration-dependent manner, with an approximate EC50 of 1.8 × 10−8 M, whereas other steroid derivatives had no effects. GC contractility needed de novo protein synthesis. The GC-induced increase in contractility was found to be consistent with an increase in F-actin content. In conclusion, lung fibroblast contractility was enhanced with GCs through an upregulation of lung fibroblast F-actin.

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Differential regulation of metalloproteinase production, proliferation and chemotaxis of human lung fibroblasts by PDGF, Interleukin-1β and TNF-α

Mediators of Inflammation ◽

10.1080/09629350020002895 ◽

2000 ◽

Vol 9 (3-4) ◽

pp. 155-160 ◽

Cited By ~ 64

Author(s):

Masahiro Sasaki ◽

Masayuki Kashima ◽

Takefumi Ito ◽

Akiko Watanabe ◽

Noriko Izumiyama ◽

...

Keyword(s):

Matrix Metalloproteinase ◽

Human Lung ◽

Lung Fibroblast ◽

Differential Sensitivity ◽

Lung Fibroblasts ◽

Type I ◽

Dependent Manner ◽

Human Lung Fibroblast ◽

Human Lung Fibroblasts ◽

Tnf Α

Fibroblast migration, proliferation, extracellular matrix protein synthesis and degradation,all of which play important roles in inflammation, are them selves induced by various growth factors and cytokines. Less is known about the interaction of these substances on lung fibroblast function in pulmonary fibrosis.The goal of this study was to investigate the effects of PDGF alone and in combination with IL–1β and TNF–α on the production of human lung fibroblast matrix metalloproteinases, proliferation, and the chemotactic response. The assay for MMPs activity against FITC labeled type I and IV collagen was based on the specificity of the enzyme cleavage of collagen. Caseinolytis and gelatinolytic activities of secreted proteinases were analyzed by zymography. Fibronectin in conditioned media was measured using human lung fibronectin enzyme immunoassay. Cell proliferation was measured by 3H-Thymidine incorporation assay. Cell culture supernatants were tested for PGE2 content by ELISA. Chemotactic activity was measured using the modified Boyden chamber.Matrix metalloproteinase assay indicated that IL–1β, TNF–α and PDGF induced intestitial collagenase (MMP-1) production. MMP assay also indicated that IL–1β and TNF–α had inhibitory effects on MMP-2,9(gelatinaseA,B) production. Casein zymography confirmed that IL–1β stimulated stromlysin (matrix metalloproteinase 3; MMP–3) and gelatin zymography demonstrated that TNF–α induced MMP–9 production in human lung fibroblast, whereas PDGF alone did not. PDGF in combination with IL–1β and TNF–α induced MMP–3 and MMP–9 activity, as demonstrated by zymography. PDGF stimulated lung fibroblast proliferation in a concentration-dependent manner, whereas IL–1β and TNF–α alone had no effect. In contrast, the proliferation of human lung fibroblasts by PDGF was inhibited in the presence of IL–1β and TNF–α, and this inhibition was not a consequence of any elevation of PGE2. PDGF stimulated fibroblast chemotaxis in a concentrationdependent manner, and this stimulation was augmented by combining PDGF with IL–1β and TNF–α.These findings suggested that PDGF differentially regulated MMPs production in combination with cytokines, and further that MMP assay and zymography had differential sensitivity for detecting MMPs. The presence of cytokines with PDGF appears to modulate the proliferation and chemotaxis of human lung fibroblasts.

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PGE2Desensitizesβ-Agonist Effect on Human Lung Fibroblast-Mediated Collagen Gel Contraction through Upregulating PDE4

Mediators of Inflammation ◽

10.1155/2013/145197 ◽

2013 ◽

Vol 2013 ◽

pp. 1-9 ◽

Cited By ~ 1

Author(s):

Qiuhong Fang ◽

Yingmin Ma ◽

Jing Wang ◽

Joel Michalski ◽

Stephen I. Rennard ◽

...

Keyword(s):

Human Lung ◽

Collagen Gel ◽

Inhibitory Effect ◽

Lung Fibroblast ◽

Lung Fibroblasts ◽

Pde4 Inhibitor ◽

Dependent Manner ◽

Human Lung Fibroblast ◽

Collagen Gel Contraction ◽

Long Acting

In the current study, we investigated the effect of a long-actingβ-agonist (salmeterol) and a phosphodiesterase 4 (PDE4) inhibitor (cilomilast) on human lung fibroblast-mediated collagen gel contraction. Higher concentrations of salmeterol (10−7and 10−6 M) inhibited fibroblast-mediated collagen gel contraction. No effect was observed with cilomilast alone (up to 10−5 M). In the presence of 10−8 M salmeterol, however, cilomilast could significantly inhibit fibroblast-mediated collagen gel contraction in a concentration-dependent manner (10−7~10−5 M). Blockade of endogenous PGE2by indomethacin further potentiated the inhibitory effect of salmeterol on fibroblast-mediated collagen gel contraction, but it did not affect cilomilast's effect. Pretreatment with PGE2abolished the inhibitory effect of salmeterol, but it potentiated the inhibitory effect of cilomilast on fibroblast-mediated collagen gel contraction. Finally, indomethacin slightly inhibited PDE4C expression, while PGE2stimulated the expression of PDE4A and -4C in human lung fibroblasts. These findings suggest that long-actingβ-agonist and PDE4 inhibitor have a synergistic effect in regulating fibroblast tissue repair functions and that PGE2can modulate the effect ofβ-agonist and PDE4 inhibitor at least in part through the mechanism of regulating PDE4 expression.

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Cyclic Mechanical Deformation Stimulates Human Lung Fibroblast Proliferation and Autocrine Growth Factor Activity

American Journal of Respiratory Cell and Molecular Biology ◽

10.1165/ajrcmb/9.2.126 ◽

1993 ◽

Vol 9 (2) ◽

pp. 126-133 ◽

Cited By ~ 42

Author(s):

Jill E. Bishop ◽

John J. Mitchell ◽

P. Marlene Absher ◽

Linda Baldor ◽

Henry A. Geller ◽

...

Keyword(s):

Growth Factor ◽

Human Lung ◽

Mechanical Deformation ◽

Lung Fibroblast ◽

Fibroblast Proliferation ◽

Factor Activity ◽

Autocrine Growth ◽

Human Lung Fibroblast ◽

Autocrine Growth Factor

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Surfactant downregulates synthesis of DNA and inflammatory mediators in normal human lung fibroblasts

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.1996.270.1.l159 ◽

1996 ◽

Vol 270 (1) ◽

pp. L159-L163 ◽

Cited By ~ 3

Author(s):

M. J. Thomassen ◽

J. M. Antal ◽

B. P. Barna ◽

L. T. Divis ◽

D. P. Meeker ◽

...

Keyword(s):

Inflammatory Cytokines ◽

Inflammatory Mediators ◽

Human Lung ◽

Thymidine Incorporation ◽

Interleukin 1 ◽

S Phase ◽

Lung Fibroblast ◽

Lung Fibroblasts ◽

Human Lung Fibroblast ◽

Normal Human

The initial inflammatory event in the adult respiratory distress syndrome (ARDS) is followed by fibroproliferation and a cascade of fibroblast-derived mediators. Because lung fibroblasts may be exposed to surfactant as well as inflammatory cytokines during ARDS, we hypothesized that surfactant might modulate fibroblast activity. We previously demonstrated that surfactant inhibited production of inflammatory cytokines from endotoxin-stimulated human alveolar macrophages. In the current study the effects of surfactant on normal human lung fibroblast proliferative capacity and mediator production were examined. Both synthetic (Exosurf) and natural (Survanta) surfactant inhibited fibroblast [3H]thymidine incorporation. Examination of pre-S-phase events indicated stimulation of the immediate response gene, c-fos, and no effect on the G1/S cyclin, cyclin D1, suggesting that the surfactant block occurred elsewhere before S phase. The antioxidant N-acetyl-L-cysteine (NAC), like surfactant, inhibited [3H]thymidine incorporation. Furthermore, menadione, a generator of intracellular H2O2, stimulated fibroblast [3H]thymidine incorporation, and this was inhibited by surfactant. Interleukin-1 (IL-1)-stimulated secretion of the inflammatory mediators, IL-6 and prostaglandin E2, was also inhibited by surfactant. These data suggest that surfactant may modify lung fibroblast participation in ARDS sequelae by downregulating DNA synthesis and secondary inflammatory mediator production.

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Effects of Platelet-Derived Growth Factor Isoforms on Human Lung Fibroblast Proliferation and Procollagen Gene Expression

Experimental Lung Research ◽

10.3109/01902149309064350 ◽

1993 ◽

Vol 19 (3) ◽

pp. 327-344 ◽

Cited By ~ 30

Author(s):

Joan G. Clark ◽

David K. Madtes ◽

Ganesh Raghu

Keyword(s):

Gene Expression ◽

Growth Factor ◽

Human Lung ◽

Lung Fibroblast ◽

Platelet Derived Growth Factor ◽

Fibroblast Proliferation ◽

Human Lung Fibroblast

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MAPK pathway mediates muscarinic receptor-induced human lung fibroblast proliferation

Life Sciences ◽

10.1016/j.lfs.2007.02.027 ◽

2007 ◽

Vol 80 (24-25) ◽

pp. 2259-2262 ◽

Cited By ~ 38

Author(s):

Sonja Matthiesen ◽

Amit Bahulayan ◽

Olaf Holz ◽

Kurt Racké

Keyword(s):

Muscarinic Receptor ◽

Human Lung ◽

Mapk Pathway ◽

Lung Fibroblast ◽

Fibroblast Proliferation ◽

Human Lung Fibroblast

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Effect of heparin and related glycosaminoglycan on PDGF-induced lung fibroblast proliferation, chemotactic response and matrix metalloproteinases activity

Mediators of Inflammation ◽

10.1080/096293500411541 ◽

2000 ◽

Vol 9 (2) ◽

pp. 85-91 ◽

Cited By ~ 16

Author(s):

Masahiro Sasaki ◽

Masayuki Kashima ◽

Takefumi Ito ◽

Akiko Watanabe ◽

Masaaki Sano ◽

...

Keyword(s):

Pulmonary Fibrosis ◽

Human Lung ◽

Heparan Sulphate ◽

Inhibitory Effect ◽

Lung Fibroblast ◽

Chemotactic Response ◽

Fibroblast Proliferation ◽

Human Lung Fibroblast ◽

Fibroblast Migration

Fibroblast migration, proliferation, extacellular matrix protein synthesis and degradation are the key events in various biological and pathological processes in pulmonary fibrosis. In addition, biopsy specimens from the lungs of patients with plumomary fibrosis show increased numbers of mast cells which have metachromatic granules containing heparin, histamin and proteases. Little is known about how these products influence pulmonary fibrosis. In the present study, we investigated the effect of heparin and related glycosaminoglycans on PDGF-induced lung fibroblast proliferation and chemotactic responsein vitro. In addition, we examined the effect of heparin on both the induction of matorix metalloproteinases (MMPs) and MMPs activity in lung fibroblastsin vitro.Heparin, de-N-sulphated heparin but not heparan sulphate inhibited PDGF-induced lung fibroblast proliferation. In contrast, only heparin inhibited PDGF-stimulated human lung fibroblast chemotaxis. Negatively charged poly-L-gultamic acid had no effect on either fibroblast proliferation or chemotaxis. Thus the negative charge alone cannot account for the ant-proliferative and anti-chemotactic effects of heparin.Furthermore, heparin and heparan sulphate also had no inhibitory effect on induction of MMPs, including MMP-1 (interstitial collagenase), MMP-2 (gelatinase A) and MMP-9 (gelatinase B). Only heparin inhibited both MMP-1 and MMP-2/MMP-9 activity. Additionally, tissue inhibitor of metalloproteinase type 1 (TIMP-1) and type 2 (TIMP-2) inhibited PDGF-stimulated human lung fibroblast chemotaxis. The ability of heparin to inhibit fibroblast chemotaxis may account for the inhibitory effect of heparin on MMP activity.The above results suggested that heparin and related glycosaminoglycans differentially regulate PDGF-induced lung fibroblast proliferation, chemotaxis and MMPs activity and further that these effects may have a key role in extracellular matrix remodeling in inflammatory lung disease.

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