Pulmonary and Critical Care Medicine Section, Departments of Internal Medicine and Pharmacology, University of Nebraska Medical Center, Omaha, Nebraska 68198-5125
![]() |
ABSTRACT |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
Bradykinin is a multifunctional mediator of
inflammation believed to have a role in asthma, a disorder associated
with remodeling of extracellular connective tissue. Using contraction
of collagen gels as an in vitro model of wound contraction, we assessed
the effects of bradykinin tissue on remodeling. Human fetal lung
fibroblasts were embedded in type I collagen gels and cultured for 5 days. After release, the floating gels were cultured in the presence of
bradykinin. Bradykinin significantly stimulated contraction in a
concentration- and time-dependent manner. Coincubation with phosphoramidon augmented the effect of 109 and
10
8 M bradykinin. A B2 receptor antagonist
attenuated the effect of bradykinin, whereas a B1 receptor
antagonist had no effect, suggesting that the effect is mediated by the
B2 receptor. An inhibitor of intracellular Ca2+
mobilization abolished the response; addition of EGTA to the culture
medium attenuated the contraction of control gels but did not modulate
the response to bradykinin. In contrast, the phospholipase C inhibitor
U-73122 and the protein kinase C inhibitors staurosporine and
GF-109203X attenuated the responses. These data suggest that by
augmenting the contractility of fibroblasts, bradykinin may have an
important role in remodeling of extracellular matrix that may result in
tissue dysfunction in chronic inflammatory diseases, such as asthma.
asthma; fibroblasts; phospholipase C; three-dimensional collagen gels
![]() |
INTRODUCTION |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
BRADYKININ IS A NONAPEPTIDE that has an important role in trauma and inflammation. Bradykinin affects vascular tone and permeability, increases secretion of mucus, and contracts smooth muscle cells (38, 46). Bradykinin has been proposed as a putative mediator of asthma because asthmatic subjects are hyperresponsive to bradykinin and because immunoreactive kinins are increased in the bronchoalveolar lavage fluids of asthmatic patients (2, 42, 43).
Rearrangement of extracellular matrix is an important aspect of both normal wound healing and fibrosis. In the airways, peribronchial fibrosis is a feature of both asthma and chronic bronchitis. This lesion may lead to the narrowing of small airways and could contribute to the fixed airflow limitation that compromises respiratory function. Fibroblasts are known to generate a traction force and to participate in tissue rearrangement (15, 39). This suggests that modulation of fibroblast contraction by inflammatory mediators like bradykinin might lead to altered tissue structure.
An in vitro model for extracellular matrix rearrangement is the
three-dimensional system of fibroblasts cultured in a native collagen
gel (3, 11, 40). When fibroblasts are cultured in such a
collagen gel, the gels are contracted by the traction force that
fibroblasts generate. Platelet-derived growth factor, transforming
growth factor-, and fetal calf serum (FCS) have been known to
augment the contraction (3, 21, 34), whereas
-adrenergic agonists and PGE2 inhibit contraction
(10, 31). In the present study, the effect of bradykinin
on fibroblast contractility was investigated with this model. The
results indicate that bradykinin can augment fibroblast-mediated gel
contraction. Bradykinin therefore may affect the fibrotic process by
augmenting fibroblast contractility and thus could contribute to the
formation of abnormal tissue architecture as well as to acute inflammation.
![]() |
METHODS |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
Materials. Type I collagen (rat tail tendon collagen, RTTC) was extracted from rat tail tendons by a previously published method (11). Briefly, tendons were excised from rat tails, and the tendon sheath and other connective tissues were removed carefully. After repeated washing with phosphate-buffered saline (GIBCO BRL) and 95% ethanol, type I collagen was extracted in 4 mM acetic acid. Protein concentration was determined by weighing a lyophilized aliquot from each lot of collagen solution. SDS-PAGE routinely demonstrated no detectable proteins other than type I collagen.
Bradykinin, D-Arg-[Hyp3,Thi5,8,D-Phe7]-bradykinin, NFibroblasts. Human fetal lung fibroblasts (HFL1) and human bronchial fibroblasts (HBF; ATCC CCD-14Br) were obtained from the American Type Culture Collection (Manassas, VA). The cells were cultured in 100-mm tissue culture dishes (Falcon, Becton Dickinson Labware, Lincoln Park, NJ) with Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% FCS, 50 U/ml penicillin, 50 µg/ml streptomycin, and 2.5 µg/ml Fungizone. The fibroblasts were usually passaged weekly. Subconfluent fibroblasts were trypsinized (trypsin-EDTA; 0.05% trypsin and 0.5 mM EDTA-4Na) and used for collagen gel culture. Fibroblasts used in these experiments were between cell passages 15 and 20 (HFL1) or 5 and 8 (HBF).
Preparation of collagen gels. Collagen gels were prepared as described previously (32). Briefly, RTTC, distilled water, 4× concentrated DMEM, and cell suspensions were mixed so that the final mixture resulted in 0.75 mg/ml of collagen, 3 × 105 cells/ml, a physiological ionic strength, and 1× DMEM. Then 500-µl aliquots of the mixture (1.5 × 105 cells/gel) were put into each well of 24-well tissue culture plates (Falcon). Gelation was generally completed within 10 min at 37°C. After gelation, 500 µl of serum free culture medium were placed over the collagen gels. The overlayered medium was changed every other day. Ham's F-12-DMEM (GIBCO BRL) was supplemented with hydrocortisone (0.1 µM), insulin (2.5 µg/ml), bovine pituitary extract (0.25%), epidermal growth factor (2.5 ng/ml), and transferrin (5 µg/ml) and used as a serum-free culture medium (4).
Rapid contraction assay. The effect of bradykinin on the fibroblast-mediated gel contraction was examined by a modification of a previously published method (32, 40). Briefly, fibroblasts were cultured in collagen gels for 5 days. After being washed three times with 1 ml of DMEM each for a 30-min incubation, the collagen gels were released from the tissue culture plates with a scalpel and transferred to 60-mm tissue culture dishes (Falcon), which contained 5 ml of DMEM with or without bradykinin and other reagents. The gels were then incubated at 37°C under a 5% CO2 atmosphere for the indicated period. Various reagents were added before release or simultaneously with release as described separately. The contraction of collagen gels was quantified by measuring the area of gels using an Optomax V image analyzer (Analytical Measuring Systems, Essex, England). Data are expressed as the percentage of each gel compared with the area measured immediately after release.
Statistical evaluation. The results are expressed as means ± SE of three separate determinations determined from triplicate gels within each experiment. Inasmuch as the number of cell passages, the batch of RTTC, and culture conditions can affect the gel contraction, all data shown in each figure were taken from a single experiment. Results, however, were always confirmed by repeating each experiment on separate occasions at least three times. The data were analyzed using an unpaired two-tailed Student's t-test and two-way ANOVA with repeated measures. Comparisons were considered statistically significant at P < 0.05.
![]() |
RESULTS |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
Time- and bradykinin concentration-dependent augmentation of
fibroblast-mediated contraction of collagen gel.
Bradykinin significantly augmented fibroblast-mediated collagen gel
contraction in a time- and concentration-dependent manner (Figs.
1 and 2).
There was no detectable contraction of the control gels in 30 min.
After 60 min, control gels had contracted 10.0 ± 1.4%, and they
continued to contract throughout the 360-min observation period. In
contrast, with 109 or 10
7 M bradykinin,
there was detectable contraction after 30 min of incubation.
Contraction at 30 min, as a percent decrease in area, was 13.1 ± 1.7% with 10
9 M bradykinin and 26.0 ± 1.6% with
10
7 M. After 60-min incubation, the contraction was
21.3 ± 0.3% with 10
9 M bradykinin and 32.2 ± 1.6% with 10
7 M (Fig. 1). Augmented contraction could be
observed with 10
7 M bradykinin throughout the 360-min
incubation. The difference between 10
9 M bradykinin and
control, observable after 30 min, was diminished by 360 min.
Bradykinin-augmented contraction was concentration dependent over the
entire range of 10
10 to 10
6 M tested (Fig.
2). Because a "plateau" was not observed, it was not possible to
calculate an EC50 (the concentration of bradykinin at which
a 50% of maximal effect of bradykinin is induced). To determine
whether adult airway fibroblasts could respond to bradykinin, HBF were
similarly tested. Bradykinin (10
7 M) resulted in
augmented contraction (83.6 ± 0.8% of original size) compared
with control (95.4 ± 1.1% of original size, P < 0.01).
|
|
Effect of protease inhibitors on bradykinin augmented fibroblast
gel contraction.
Peptidase degradation of small peptides might be an important
regulatory process in vivo (27, 42). Because fibroblasts are known to have two membrane-bound peptidases, neutral endopeptidase (NEP) (24, 28) and angiotensin-converting enzyme (ACE)
(37, 45), which are capable of cleaving bradykinin, the
effects of the NEP inhibitors phosphoramidon and thiorphan and the ACE
inhibitor captopril (9) were evaluated. The effect of
109 and 10
8 M bradykinin on fibroblast gel
contraction was significantly increased in the presence of 1 µM
phosphoramidon (Fig. 3, P < 0.01). In contrast, captopril showed no effect, and the addition of
captopril to phosphoramidon did not further augment contraction. Thiorphan (10 µM) also augmented the effect of bradykinin similarly to the effect of phosphoramidon (data not shown).
|
Effect of B1 and B2 antagonists.
The effect of bradykinin is mediated by specific receptors, which
belong to two major categories, B1 and B2
(7, 35). To evaluate which receptor mediates the effect of
bradykinin on fibroblast gel contraction, the effects of a
B1 receptor competitive antagonist
des-Arg9,[Leu8]-bradykinin and
B2 receptor competitive antagonists
D-Arg-[Hyp3,Thi5,8, D-Phe7]-bradykinin
and
N-adamantaneacetyl-D-Arg-[Hyp3,Thi5,8,D-Phe7]-bradykinin
(26) were evaluated. As shown in Fig.
4, the B2 antagonist
N
-adamantaneacetyl-D-Arg-[Hyp3,Thi5,8,D-Phe7]-bradykinin
attenuated the effect of bradykinin (P < 0.01 by ANOVA), whereas the B1 antagonist
des-Arg9,[Leu8]-bradykinin had no effect. The
B2 antagonist
D-Arg-[Hyp3,Thi5,8, D-Phe7]-bradykinin
also inhibited the effect of bradykinin (data not shown). Thus the
B2 receptor appears to mediate the enhancing effect of
bradykinin on fibroblast contraction of collagen gels.
|
Signal transduction pathways mediating the effects of bradykinin.
Because the effects of bradykinin in some cells are known to be
mediated by a PTX-sensitive G protein (6, 44), the effect of PTX on bradykinin-augmented fibroblast-mediated collagen gel contraction was investigated. PTX treatment did not affect the fibroblast-mediated collagen gel contraction in either the absence or
the presence of bradykinin (Fig. 5).
|
|
|
|
![]() |
DISCUSSION |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
The current study demonstrates that bradykinin augments fibroblast-mediated collagen gel contraction. Addition of a NEP inhibitor further augmented the effect of bradykinin. The B2 receptor appeared to mediate the effect because a B2 receptor competitive antagonist attenuated the response, whereas a B1-receptor competitive antagonist did not. The PKC inhibitors staurosporine and GF-109203X and the PLC inhibitor U-73122 inhibited control and bradykinin-induced contraction. The Ca2+ antagonist TMB-8 abolished the bradykinin-stimulated response. PTX treatment and the addition of EGTA to the culture medium failed to modulate the response to bradykinin. Thus the effect of bradykinin seemed to depend on PLC and PKC activation and intracellular Ca2+ mobilization.
Rearrangement of extracellular matrix is a crucial process in both wound healing and fibrosis. One aspect of the rearrangement of extracellular connective tissue matrix is contraction. The contraction of wounded tissue can minimize the area to be covered by epithelium and may promote healing. The contraction of tissue and matrix in chronic diseases, however, may lead to disruption of tissue architecture and cause tissue dysfunction (12, 15, 17). Whereas the mechanisms involved in the contraction of fibrotic tissues during repair processes are not fully described, fibroblasts can generate a traction force and can participate in this process (22, 39). When fibroblasts are cultured in a three-dimensional native type I collagen gel, they contract the collagen gel. This phenomenon has been considered to be a model of tissue rearrangement (17). In the current study, this model was used to investigate the contractility of fibroblasts.
Bradykinin is a nonapeptide that has been implicated in the response to
trauma and injury (38, 42). Bradykinin is known to
increase vascular permeability and secretion of mucus, to cause vasodilation, and to contract smooth muscle both in vivo and in vitro
(46). Increased levels of bradykinin have been reported in
bronchoalveolar lavage fluids from asthmatic patients (2). Bradykinin also provokes bronchoconstriction in asthmatic subjects (14). Here we reported that bradykinin (1010
to 10
6 M) augmented the contraction of type I collagen
gels by HFL1 or HBF cells. The concentrations of bradykinin used in
this study are close to the bradykinin amount found in human sputum
(23). Thus bradykinin may have an important role in asthma
and other inflammatory diseases.
Peptidases that are responsible for the breakdown of bradykinin may have important roles in modulating bradykinin-induced effects (27, 42). Although many proteases are able to hydrolyze kinins, two membrane-bound enzymes, NEP and ACE, seem to have important roles (9). NEP, which is present in a variety of airway cells, can cleave many peptides including bradykinin, substance P, neurokinin A, and vasoactive intestinal peptide (24, 28). ACE also is present in a variety of cells, including fibroblasts, and can degrade bradykinin (33, 37, 45). In the current study, the NEP inhibitors phosphoramidon and thiorphan augmented the effect of bradykinin, whereas the ACE inhibitor captopril had no effect. These results suggest that the effect of bradykinin can be modulated by peptidase activity and that under the culture conditions used, NEP has a more important role than ACE.
The receptors mediating responses to bradykinin have typically been divided into two major subtypes, B1 and B2 (35). The B1 receptor is not expressed at significant levels in normal tissues, but its synthesis can be induced after tissue injury or inflammation. The B2 receptor is constitutively expressed in various kinds of cells including smooth muscle cells, certain neurons, fibroblasts, and epithelial cells and is responsible for many of bradykinin's diverse biological effects. The current study suggests that the effects of bradykinin on fibroblast-mediated collagen gel contraction are mediated by the B2 receptor. Consistent with this observation, lung fibroblasts have been demonstrated to express the B2 receptor (1, 25).
Bradykinin activates subcellular responses not only directly but also
indirectly through the release of other mediators, including arachidonic acid metabolites, nitric oxide, platelet-activating factor,
tumor necrosis factor (TNF)-, interleukin (IL)-1, norepinephrine and
neuropeptides, depending on the cell types and responses (7, 35,
42). The effects of the cyclooxygenase inhibitor indomethacin, the lipooxygenase inhibitors NDGA and Wy-50295M (16), the
nitric oxide synthesis inhibitor
NG-monomethyl-L-arginine citrate,
and the nitric oxide generator nitroprusside were evaluated.
These reagents tested all failed to modulate the augmentation of the
fibroblast-mediated collagen gel contraction induced by bradykinin
(data not shown). Thus the effects of bradykinin appear not to be
caused indirectly by eicosanoids or nitric oxide-mediated mechanisms.
Bradykinin can lead to indirect effects through other mechanisms as
well. In this regard, IL-1, TNF-
, platelet-activating factor, and
substance P were all found to be without enhancing effect on
fibroblast-mediated gel contraction in our system (data not shown),
suggesting that the bradykinin effect is not mediated indirectly
through these agonists either.
The signaling pathway(s) by which bradykinin enhances fibroblast-mediated gel contraction was investigated. Bradykinin receptors are members of the G protein-coupled receptor superfamily, and bradykinin can induce cellular effects by activation of either PTX-sensitive or -insensitive G proteins (7, 44). PTX failed to inhibit the effects of bradykinin, suggesting involvement of a PTX-insensitive G protein pathway.
One of the PTX-insensitive signaling pathways that can be activated by bradykinin in other cells is activation of the phosphoinositide-specific PLC enzyme, leading to hydrolysis of phosphatidylinositol 4,5-bisphosphate and formation of inositol 1,4,5-trisphosphate and diacylglycerol (13, 29, 30). Inositol trisphosphate leads to mobilization of intracellular calcium and diacylglycerol leads to activation of PKC. It has been reported that the stimulatory effects on fibroblast-mediated gel contraction of serum, endothelin-1, and platelet-derived growth factor are dependent on activation of PKC (8, 18-20). Our results with bradykinin are consistent with these data, although the contraction assay used by Guidry and associates (18-20) was slightly different from that used in the current study. Importantly, inhibition of PLC or PKC in the present study inhibited control gel contraction as well as bradykinin-augmented contraction. Thus it is possible that PKC and PLC are needed for gel contraction, and their role in bradykinin signal transduction in this regard is not yet established.
In conclusion, the current study demonstrates that fibroblast-mediated collagen gel contraction can be augmented by bradykinin. The ability of bradykinin to modulate this process suggests that bradykinin may have an important role in fibrotic processes. The possibility of modulating this process by targeting bradykinin receptors or the subsequent signaling pathway involved might offer a novel therapeutic opportunity in a variety of destructive and fibrotic diseases.
![]() |
ACKNOWLEDGEMENTS |
---|
We acknowledge Mary G. Illig, Art Heires, Ronald F. Etrl, and Leroy Tate for assistance with cell culture and Lillian Richards for secretarial assistance.
![]() |
FOOTNOTES |
---|
Present addresses: T. Mio, Pulmonary Medicine, Chest Disease Res. Inst., Kyoto Univ., 53 Kawamaramachi Shogoin Sukyo-ku, Kyoto, Japan 601; Y. Adachi, Dept. of Pediatrics, Toyama Med. and Pharm. Univ, 2630 Sugitani, Toyama 930-01, Japan.
Address for reprint requests and other correspondence: S. I. Rennard, Pulmonary and Critical Care Medicine Sect., Dept. of Internal Medicine, Univ. of Nebraska Medical Center, 985125 Nebraska Medical Center, Omaha, NE 68198-5125 (E-mail: srennard{at}unmc.edu).
The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
Received 27 July 2000; accepted in final form 12 February 2001.
![]() |
REFERENCES |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
1.
Baenziger, NL,
Jong YJ,
Yocum SA,
Dalemar LR,
Wilhelm B,
Vavrek R,
and
Stewart JM.
Diversity of B2 bradykinin receptors with nanomolar affinity expressed in passaged IMR90 human lung fibroblasts.
Eur J Cell Biol
58:
71-80,
1992[ISI][Medline].
2.
Baumgarten, CR,
Lehmkuhl B,
Henning R,
Burunee T,
Dorow P,
Schilling W,
and
Kunkel G.
Bradykinin and other inflammatory mediators in BAL-fluid from patients with active pulmonary inflammation.
Agents Actions Suppl
38:
475-481,
1992[Medline].
3.
Bell, E,
Ivarsson B,
and
Merrill C.
Production of a tissue-like structure by contraction of collagen lattices by human fibroblasts of different proliferative potential in vitro.
Proc Natl Acad Sci USA
76:
1274-1278,
1979[Abstract].
4.
Bottenstein, J,
Hayashi I,
Hutchings S,
Masui H,
Mather J,
McClure DB,
Ohasa S,
Rizzino A,
Sato G,
Serrero G,
Wolfe R,
and
Wu R.
The growth of cells in serum-free hormone-supplemented media.
Methods Enzymol
58:
94-109,
1979[Medline].
6.
Bueb, JL,
Mousli M,
Landry Y,
and
Bronner C.
A pertussis toxin-sensitive G protein is required to induce histamine release from rat peritoneal mast cells by bradykinin.
Agents Actions
30:
98-101,
1990[ISI][Medline].
7.
Burbach, JP,
and
Meijer OC.
The structure of neuropeptide receptors.
Eur J Pharmacol
227:
1-18,
1992[Medline].
8.
Choudhury, P,
Chen W,
and
Hunt RC.
Production of platelet-derived growth factor by interleukin-1 beta and transforming growth factor-beta-stimulated retinal pigment epithelial cells leads to contraction of collagen gels.
Invest Ophthalmol Vis Sci
38:
824-833,
1997[Abstract].
9.
Dusser, DJ,
Nadel JA,
Sekizawa K,
Graf PD,
and
Borson DB.
Neutral endopeptidase and angiotensin converting enzyme inhibitors potentiate kinin-induced contraction of ferret trachea.
J Pharmacol Exp Ther
244:
531-536,
1988[Abstract].
10.
Ehrlich, HP,
and
Wyler DJ.
Fibroblast contraction of collagen lattices in vitro: inhibition by chronic inflammatory cell mediators.
J Cell Physiol
116:
345-351,
1983[ISI][Medline].
11.
Elsdale, T,
and
Bard J.
Collagen substrata for studies on cell behavior.
J Cell Biol
54:
626-637,
1972
12.
Evans, JN,
Kelley J,
Low RB,
and
Adler KB.
Increased contractility of isolated lung parenchyma in an animal model of pulmonary fibrosis induced by bleomycin.
Am Rev Respir Dis
125:
89-94,
1982[ISI][Medline].
13.
Field, JL,
Butt SK,
Morton IKM,
and
Hall JM.
Bradykinin B2 receptors and coupling mechanisms in the smooth muscle of the guinea-pig taenia caeci.
Br J Pharmacol
113:
607-613,
1994[Abstract].
14.
Fuller, RW,
Dixon CMS,
Cuss FMC,
and
Barnes PJ.
Bradykinin-induced bronchoconstriction in humans: mode of action.
Am Rev Respir Dis
135:
176-180,
1987[ISI][Medline].
15.
Gabbiani, G,
Hirschel BJ,
Ryan GB,
Statkov PR,
and
Maino G.
Granulation tissue as a contractile organ: a study of structure and function.
J Exp Med
135:
719-733,
1972[ISI][Medline].
16.
Grimes, D,
Sturm RJ,
Marinari LR,
Carlson RP,
Berkenkopf JW,
Musser JH,
Kreft AF,
and
Weichman BM.
WY-50,295 tromethamine, a novel, orally active 5-lipoxygenase inhibitor: biochemical characterization and antiallergic activity.
Eur J Pharmacol
236:
217-228,
1993[ISI][Medline].
17.
Grinnell, F.
Fibroblasts, myofibroblasts and wound contraction.
J Cell Biol
124:
401-404,
1994[ISI][Medline].
18.
Guidry, C.
Extracellular matrix contraction by fibroblasts: peptide promoters and second messengers.
Cancer Metastasis Rev
11:
45-54,
1992[ISI][Medline].
19.
Guidry, C.
Fibroblast contraction of collagen gels requires activation of protein kinase C.
J Cell Physiol
155:
358-367,
1993[ISI][Medline].
20.
Guidry, C,
and
Hardwick C.
Extracellular matrix contraction by choroidal fibroblasts: inhibition by staurosporine.
Invest Ophthalmol Vis Sci
35:
503-508,
1994[Abstract].
21.
Gullberg, D,
Tingstrom A,
Thuresson A-C,
Olsson L,
Terracio L,
Borg TK,
and
Rubin K.
1 integrin-mediated collagen gel contraction is stimulated by PDGF.
Exp Cell Res
186:
264-272,
1990[ISI][Medline].
22.
Harris, AK.
Fibroblast traction as a mechanism for collagen morphogenesis.
Nature
290:
249-251,
1981[ISI][Medline].
23.
Ichinose, M,
Takahashi T,
Sugiura H,
Endoh N,
Miura M,
Mashito Y,
and
Shirato K.
Baseline airway hyperresponsiveness and its reversible component: role of airway inflammation and airway calibre.
Eur Respir J
15:
248-253,
2000
24.
Johnson, AR,
Ashton J,
Schulz WW,
and
Erdoes EG.
Neutral metalloendopeptidase in human lung tissue and cultured cells.
Am Rev Respir Dis
132:
564-568,
1985[ISI][Medline].
25.
Jong, YJ,
Dalemar LR,
Wilhelm B,
and
Baenziger NL.
Human bradykinin B2 receptors isolated by receptor-specific monoclonal antibodies are tyrosine phosphorylated.
Proc Natl Acad Sci USA
90:
10994-10998,
1993[Abstract].
26.
Lammek, B,
Wang YK,
Gavras I,
and
Gavras H.
A novel bradykinin antagonist with improved properties.
J Pharm Pharmacol
43:
887-888,
1991[ISI][Medline].
26a.
Lechner, JF,
and
LaVeck MA.
A serum-free method for culturing normal human bronchial epithelial cells at clonal density.
J Tissue Cult Methods
9:
43-45,
1985.
27.
Lilly, CM,
Drazen JM,
and
Shore SA.
Peptidase modulation of airway effects of neuropeptides.
Proc Soc Exp Biol Med
203:
388-404,
1993[Abstract].
28.
Lorkowski, G,
Zijderhand-Bleekemolen E,
Erdoes EG,
and
von Figura K.
Neutral endopeptidase-24.11 (enkephalinase): biosynthesis and localization in human fibroblasts.
Biochem J
248:
345-350,
1987[ISI][Medline].
29.
Marsh, KA,
and
Hill SJ.
Bradykinin B2 receptor-mediated phophoinositide hydrolysis in bovine tracheal smooth muscle cells.
Br J Pharmacol
107:
443-447,
1992[Abstract].
30.
Marsh, KA,
and
Hill JS.
Characteristics of the bradykinin-induced changes in intracellular calcium ion concentration of single bovine tracheal smooth cells.
Br J Pharmacol
110:
29-35,
1993[Abstract].
31.
Mio, T,
Adachi Y,
Carnevali S,
Romberger DJ,
Spurzem JR,
and
Rennard SI.
-Adrenergic agonists attenuate fibroblast-mediated contraction of released collagen gels.
Am J Physiol Lung Cell Mol Physiol
270:
L829-L835,
1996
32.
Mio, T,
Adachi Y,
Romberger DJ,
Ertl RF,
and
Rennard SI.
Regulation of fibroblast proliferation in three dimensional collagen gel matrix.
In Vitro Cell Dev Biol
32:
427-433,
1996[ISI].
33.
Momose, N,
Fukuo K,
Morimoto S,
and
Ogihara T.
Captopril inhibits endothelin-1 secretion from endothelial cells through bradykinin.
Hypertension
21:
921-924,
1993[Abstract].
34.
Montesano, R,
and
Orci L.
Transforming growth factor stimulates collagen-matrix contraction by fibroblasts: implication for wound healing.
Proc Natl Acad Sci USA
85:
4894-4897,
1988[Abstract].
35.
Regoli, D,
Jukic D,
Gobeil F,
and
Rhaleb NE.
Receptors for bradykinin and related kinins: a critical analysis.
Can J Physiol Pharmacol
71:
556-567,
1993[ISI][Medline].
36.
Sako, T,
Tauber AI,
Jeng AY,
Yuspa SH,
and
Blumberg PM.
Contrasting actions of staurosporine, a protein kinase C inhibitor, on human neutrophils and primary mouse epidermal cells.
Cancer Res
48:
4646-4650,
1988[Abstract].
37.
Smallridge, RC,
Gamblin GT,
and
Eil C.
Angiotensin-converting enzyme: characteristics in human skin fibroblasts.
Metabolism
35:
899-904,
1986[ISI][Medline].
38.
Stewart, JM.
The kinin system in inflammation.
Agents Actions Suppl
42:
145-157,
1993[Medline].
39.
Stopak, D,
and
Harris AK.
Connective tissue morphogenesis by fibroblast traction.
Dev Biol
90:
383-398,
1982[ISI][Medline].
40.
Tomasek, JJ,
and
Akiyama SK.
Fibroblast-mediated collagen gel contraction does not require fibronectin-alpha5 beta1 integrin interaction.
Anat Rec
234:
153-160,
1992[ISI][Medline].
41.
Toullec, D,
Pianetti P,
Coste H,
Bellevergue P,
Grand-Perret T,
Ajakane M,
Baudet V,
Boissin P,
Boursier E,
Loriolle F,
Duhamel L,
Charon D,
and
Kirilovsky J.
The bisindolylmaleimide GF 109203X is a potent and selective inhibitor of protein kinase C.
J Biol Chem
266:
15771-15781,
1991
42.
Trifilieff, A,
Da Silva A,
and
Gies JP.
Kinins and respiratory tract diseases.
Eur Respir J
6:
576-587,
1993[Abstract].
43.
Varonier, HS,
and
Panzani R.
The effect of inhalations of bradykinin on healthy and atopic (asthmatic) children.
Int Arch Allergy
34:
293-296,
1968.
44.
Venema, VJ,
Ju H,
Sun J,
Eaton DC,
Marrero MB,
and
Venema RC.
Bradykinin stimulates the tyrosine phosphorylation and bradykinin B2 receptor association of phospholipase C gamma 1 in vascular endothelial cells.
Biochem Biophys Res Commun
246:
70-75,
1998[ISI][Medline].
45.
Weinberg, KS,
Douglas WHJ,
MacNamee DR,
Lanzillo JJ,
and
Fanburg BL.
Angiotensin I-converting enzyme localization on cultured fibroblasts by immunofluorescence.
In Vitro
18:
400-406,
1982[ISI][Medline].
46.
Wilhelm, DL.
Kinins in human disease.
Annu Rev Med
22:
63-84,
1971[ISI][Medline].