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© 1995 American Heart Association, Inc.
Articles |
Adrenomedullin as a Novel Antimigration Factor of Vascular Smooth Muscle Cells
From the First Department of Internal Medicine, Osaka (Japan) City University Medical School.
Correspondence to Takeshi Horio, MD, Division of Hypertension and Atherosclerosis, First Department of Internal Medicine, Osaka City University Medical School, 1-5-7 Asahi-machi, Abeno-ku, Osaka 545, Japan.
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Abstract |
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 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
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Abstract The present study investigated the effect of adrenomedullin, a novel vasorelaxant peptide, on the migration of cultured rat vascular smooth muscle cells (SMCs) by using the Boyden-chamber method. Fetal calf serum (FCS) and platelet-derived growth factor (PDGF)–BB strongly stimulated SMC migration. Adrenomedullin clearly inhibited SMC migration stimulated with 5% and 10% FCS in a concentration-dependent manner. The migration induced by 10 and 25 ng/mL PDGF-BB was also inhibited by adrenomedullin in a concentration-dependent manner. Inhibition by adrenomedullin of FCS- and PDGF-induced SMC migration was paralleled by an increase in the cellular level of cAMP. In fact, the percent increase in cAMP level was strongly correlated with the percent decrease in migration activity of SMCs after treatment with adrenomedullin. 8-Bromo cAMP, a cAMP analogue, reproduced the inhibition by adrenomedullin of FCS- and PDGF-induced SMC migration. An activator of adenylate cyclase, forskolin, also reduced FCS- and PDGF-induced SMC migration. These data indicate that adrenomedullin inhibits the migration of SMCs stimulated with FCS and PDGF, probably through a cAMP-dependent process. On the basis of these results and the finding that adrenomedullin is synthesized in and secreted from vascular endothelial cells, adrenomedullin may play a role as a local antimigration factor in some pathophysiological states.
Key Words: adrenomedullin • migration • smooth muscle cells
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Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Vascular endothelial cells and SMCs play important roles in atherogenesis. Migration of arterial medial SMCs into the intimal layer is a key process in intimal thickening in atherosclerotic lesions.1 2 A number of factors are known to stimulate SMC migration, such as PDGF,3 interleukin-1,4 transforming growth factor-ß,5 fibronectin,6 fibrinogen,7 and oxidized low-density lipoprotein.8 These factors are derived from blood components or secreted by vascular cells, including endothelial cells.
Vascular endothelial cells also secrete several kinds of vasoconstrictive or vasorelaxant agents. Endothelin, a potent endothelial cell–derived vasoconstrictor,9 is mitogenic for SMCs.10 In contrast, nitric oxide and C-type natriuretic peptide, which are endothelial cell–derived vasodilators,11 12 have been shown to exert antiproliferative effects on SMCs.13 14 Taken together, vascular endothelial cells may take part in the modulation of atherogenesis as well as the regulation of vascular tone by releasing various vasoactive factors.
A very recent study15 has demonstrated that adrenomedullin, a novel peptide originally isolated from human pheochromocytoma,16 is produced and released from cultured vascular endothelial cells. This 52–amino acid peptide shows slight homology with calcitonin gene–related peptide and increases cAMP in rat platelets.16 Intravenous administration of adrenomedullin is found to elicit a potent and long-lasting hypotensive effect attributable to a vascular resistance reduction in anesthetized rats.17 Furthermore, it is shown that rat vascular SMCs have specific binding sites for adrenomedullin.18 19 Therefore, adrenomedullin secreted from endothelial cells is deduced to act as a paracrine regulator in the local control of vascular tone and SMC function. It is still unclear, however, whether adrenomedullin has biological actions on SMCs other than vasorelaxation. There are no data about its effect on various events during atherogenesis in SMCs. Therefore, we conducted such a study, examining the effect of adrenomedullin on the migration of cultured rat aortic SMCs and investigating the relation between the change in SMC migration by adrenomedullin and the level of cAMP in cells.
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Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Materials
PDGF (recombinant AA, AB, and BB), 3-isobutyl-1-methylxanthine, 8-bromo cAMP, and BSA were purchased from Sigma Chemical Co. Rat adrenomedullin was purchased from Peptide Institute. Type I collagen was purchased from Koken Inc. DMEM, trypsin, penicillin, streptomycin, EDTA, and FCS were purchased from GIBCO Laboratories. Flasks and multiwell plates were purchased from Becton Dickinson & Co. The cAMP assay kit was purchased from Yamasa Shoyu Co, Ltd. Diff-Quick staining solution was purchased from Green-Cross Corp. Forskolin was provided as a gift by Nihon Kayaku Co.
Culture of SMCs
Rat vascular SMCs were grown from the aortic explants of
Sprague-Dawley rats and were cultured in DMEM containing 10%
FCS, penicillin (50 U/mL), and streptomycin (50 µg/mL) as previously
described.20 Cells were identified as SMCs according to
their morphological and immunocytochemical
characteristics.21 Briefly, these cells showed a typical
"hill-and-valley" growth pattern and had positive
fluorescence with antibodies against 
-smooth muscle
actin but negative fluorescence against factor VIII antigen.
Cultures were maintained at 37°C with atmospheric air and 5%
CO2. Cells were subcultured after treatment with 0.25%
trypsin and 0.02% EDTA. Subconfluent SMCs in their third to seventh
passages were used for the experiments.
Migration Assay
Migration of SMCs was assayed by a modification of the
Boyden-chamber method using microchemotaxis chambers (Neuro Probe
Inc) and polycarbonate filters (Nucleopore Corp) with pores of 5.0-µm
diameter, as previously reported by Koyama et al.22 In all
experiments, collagen-coated filters were used. Briefly, the
membranes were treated with 0.5N acetic acid and then incubated for 48
to 72 hours at 25°C in a collagen solution (100 µg/mL type I
collagen in 0.5N acetic acid). They were then air-dried. Cultured
SMCs were trypsinized and suspended at a concentration of
1.5x105 cells per milliliter in DMEM supplemented with
0.4% BSA. The cell number was counted with an electronic cell counter
(model ZB1, Coulter Electronics). A 200-µL volume of SMC suspension
(3.0x104 cells) was placed in the upper chamber, and 40
µL of DMEM/0.4% BSA containing a migration factor such as FCS or
PDGF was placed in the lower chamber. The chamber was incubated at
37°C under 5% CO2 in air for 2 to 8 hours. After
incubation, the filter was removed, and the SMCs on the upper side of
the filter were scraped off. The SMCs that had migrated to the lower
side of the filter were fixed in methanol, stained with Diff-Quick
staining solution, and counted under a microscope for quantification of
SMC migration. Migration activity was expressed as the number of cells
that had migrated per high-power field (x400). In experiments to
determine the effects of adrenomedullin, 8-bromo cAMP, and forskolin on
SMC migration, these agents were added to the lower chamber before the
incubation.
Adhesion Assay
SMC adhesion to the filter was assayed under conditions
identical to the SMC migration assay by using microchemotaxis chambers
and polycarbonate filters with pores of 5.0-µm diameter that had been
precoated with type I collagen. Cultured SMCs were trypsinized and
suspended at a concentration of 5.0x104 cells per
milliliter in DMEM supplemented with 0.4% BSA. A 200-µL volume of
SMC suspension (1.0x104 cells) was placed in the upper
chamber, and 40 µL of DMEM/0.4% BSA containing FCS, PDGF,
adrenomedullin, or a combination of these factors was placed in the
lower chamber. The chamber was incubated at 37°C under 5%
CO2 in air for 4 hours. After incubation, the filter was
removed and gently washed to remove nonattached cells. The adherent
SMCs on the upper side of the filter were fixed in methanol, stained
with Diff-Quick staining solution, and counted under a microscope
(x400) for quantification of SMC adhesion.
cAMP Measurement
After preincubation, cells grown in multiwell plates were washed
twice with serum-free medium and were then stimulated mainly for 30
minutes with various concentrations of rat adrenomedullin dissolved in
DMEM with 0.5 mmol/L 3-isobutyl-1-methylxanthine, a phosphodiesterase
inhibitor, as described previously.23 24 The
reaction was stopped by rapid aspiration and the addition of 2 mL of
ice-cold 65% ethanol. After evaporation by a centrifugal
evaporator, the dry residue was dissolved in an assay buffer according
to the reported method in our laboratory.25 The cAMP
levels were determined by radioimmunoassay performed with the cAMP
assay kit.
Calculations and Statistical Analysis
The statistical significance of differences in the results was
evaluated by using an unpaired ANOVA, and P values were
calculated by Scheffé's method.26 A level of
P<.05 was accepted as statistically significant.
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Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Effects of FCS and PDGF on SMC Migration
FCS and PDGF-BB stimulated the migration of SMCs in a time-dependent manner (Table 1
). SMC migration
induced by these agents increased during the initial 4 hours of
incubation, after which the rate of increase showed a significant
decline. Therefore, the subsequent studies on SMC migration were
performed with cells incubated for 4 hours.
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FCS (1% to 10%) and PDGF-BB (5 to 25 ng/mL) strongly stimulated SMC migration in a dose-dependent manner (data not shown). The stimulatory effect of PDGF-AB on SMC migration was much weaker than that of PDGF-BB, and PDGF-AA did not induce SMC migration at all (data not shown), as previously described by Koyama and colleagues.27 28 In later experiments, stimulation of SMC migration was performed with 5% or 10% FCS and 10 or 25 ng/mL PDGF-BB.
Effect of Adrenomedullin on SMC Migration Stimulated With FCS
and PDGF
The effect of adrenomedullin on the migration of SMCs treated with
FCS is shown in Fig 1. Adrenomedullin potently inhibited
SMC migration after stimulation with both 5% and 10% FCS. This
inhibition was concentration dependent. The effect of adrenomedullin on
the migration of SMCs treated with PDGF-BB is shown in Fig 2. The inhibitory effect of adrenomedullin
on SMC migration after stimulation with PDGF-BB was essentially the
same as that after stimulation with FCS. Adrenomedullin clearly
inhibited PDGF-BB (10 and 25 ng/mL)–induced SMC migration in a
concentration-dependent manner. Although nonstimulated SMCs
exhibited a little migration activity, adrenomedullin
(10-7 and 10-6 mol/L) did not inhibit this
basal activity (Table 2).
|
) or 10% (
)
FCS. Migration activities are expressed as the number of cells per
high-power field (HPF), and the values are given as the mean±SD of
four measurements. *Significant difference compared with FCS alone.
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) or 25 ng/mL
(
) PDGF-BB. Migration activities are expressed as the number of
cells per high-power field (HPF), and the values are given as the
mean±SD of four measurements. 
Significant difference compared with
PDGF-BB alone.
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Effect of Adrenomedullin on SMC Adhesion
The effect of adrenomedullin on the adhesion of SMCs to the filter
in the absence or presence of FCS or PDGF-BB is shown in Fig 3. Adrenomedullin (10-6 mol/L) had no
significant effect on cell adhesion, either in the absence or presence
of 10% FCS or 25 ng/mL PDGF-BB.
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Effect of Adrenomedullin on Cellular cAMP Level in SMCs Treated
With FCS and PDGF
In the presence of 5% FCS or 10 ng/mL PDGF-BB, cellular cAMP
levels rapidly increased after treatment of the cells with
adrenomedullin, and the increased cAMP levels were sustained over 4
hours (Fig 4A). The increase in cellular cAMP levels
induced by adrenomedullin for 30 minutes was dose dependent (Fig 4B).
Furthermore, there was a significant correlation between the percent
increase in cellular cAMP level and the percent decrease in migration
activity (Fig 5).
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Effect of 8-Bromo cAMP and Forskolin on SMC Migration Stimulated
With FCS and PDGF
To elucidate whether the inhibitory effect of
adrenomedullin on the migration of SMCs after stimulation with FCS and
PDGF is causally linked to the increase in cellular cAMP, we examined
the effect of 8-bromo cAMP, a cAMP analogue, on SMC migration treated
with FCS and PDGF-BB. Inhibition of FCS-induced and PDGF-BB–induced
SMC migration by adrenomedullin could be reproduced by this analogue at
concentrations of 10-4 and 10-3 mol/L (Fig 6). Furthermore, the effect of forskolin, an
activator of adenylate cyclase, on SMC
migration treated with FCS and PDGF-BB was examined. The addition of
forskolin also reduced FCS-induced and PDGF-BB–induced SMC migration
in a dose-dependent manner (Fig 7).
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Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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The present study has demonstrated for the first time that adrenomedullin inhibits the migration of SMCs stimulated with FCS and PDGF in a concentration-dependent manner. FCS-induced SMC migration was significantly inhibited by adrenomedullin at concentrations of 10-8 to 10-6 mol/L, and PDGF-BB–induced SMC migration was significantly inhibited by 10-9 to 10-6 mol/L adrenomedullin. Percent inhibition by 10-6 mol/L adrenomedullin of SMC migration stimulated with 10% FCS or 25 ng/mL PDGF-BB was 66% or 62%, respectively. Although it has been reported that immunoreactive adrenomedullin is present in human and rat plasma,29 30 the plasma concentrations (
10-12 to 10-11 mol/L) are much lower than
those of synthetic adrenomedullin which inhibited SMC migration
significantly in our in vitro study. However, local levels of
adrenomedullin in vascular tissues may be much higher than plasma
concentration, because it has recently been shown that a considerable
amount of adrenomedullin is secreted from vascular
endothelial cells.15 Taking this finding
into account, our results suggest that adrenomedullin, by acting
locally as a paracrine, inhibits the migration of SMCs after
stimulation with factors such as PDGF. The migration of
arterial medial SMCs into the intima is an important
process in intimal thickening not only in atherosclerotic lesions but
also in restenosis after angioplasty.2 31
Consequently, it is possible that adrenomedullin antagonizes the
development of these vascular lesions as a local antimigratory factor
for SMCs, although we have no direct evidence in vivo at this time. In the present study, adrenomedullin did not inhibit the basal migration activity of nonstimulated SMCs. Adrenomedullin did not suppress the cell adhesion either in the absence or presence of FCS or PDGF. Furthermore, in a trypan blue exclusion test, dead cells stained with trypan blue were not found 24 hours after treatment with 10-7 mol/L adrenomedullin. On the basis of these observations and the finding that cultured SMCs actively produce cAMP induced by adrenomedullin, it is unlikely that the inhibitory effect of adrenomedullin on SMC migration observed in the present study was due to its cytotoxicity.
We have obtained some evidence of a causal link between cAMP production and the inhibition of SMC migration by adrenomedullin. Adrenomedullin remarkably increased cAMP levels in cells, and this increase paralleled the inhibition of SMC migration treated with FCS or PDGF. A cAMP analogue and an activator of adenylate cyclase suppressed FCS- and PDGF-induced SMC migration. These results suggest that adrenomedullin inhibits SMC migration stimulated with FCS and PDGF, probably through a cAMP-dependent process. There are some possible mechanisms by which cAMP elevation by adrenomedullin inhibits PDGF-induced SMC migration. Compounds that increase cAMP and activate protein kinase A have been shown to inhibit the PDGF-BB–induced activation of MAP kinase and MAP kinase kinase in arterial SMCs.32 Therefore, cAMP-dependent protein kinase may mediate inhibition of PDGF-induced SMC migration by blocking MAP kinase signaling. Bornfeldt et al33 have reported that stimulation of phosphatidylinositol turnover, diacylglycerol formation, and intracellular Ca2+ flux, but not activation of the MAP kinase cascade, are likely to be required for chemotaxis of human arterial SMCs. Since cAMP decreases the intracellular Ca2+ level in SMCs,34 it is possible that the decreased cytosolic Ca2+ induced by cAMP suppresses the migratory activities of SMCs stimulated with PDGF. However, we have not elucidated the exact cellular mechanism by which cAMP inhibits FCS- and PDGF-induced migration of SMCs. As for the cell mitogenesis, cAMP elevates c-myc mRNA levels and increases DNA synthesis in Swiss 3T3 fibroblasts.35 In SMCs, conversely, cAMP inhibits serum- or PDGF-induced DNA synthesis.36 37 Therefore, cAMP shows the different biological actions on distinct cultured cell types, and so its functional mechanism is complicated. Further studies are required to clarify the exact cellular mechanism of the inhibition by adrenomedullin of SMC migration.
In conclusion, the present study indicates that adrenomedullin inhibits the migration of SMCs stimulated with FCS and PDGF-BB and that the increase in cellular cAMP level is likely to be involved in the inhibition of SMC migration by adrenomedullin. Adrenomedullin may play a role as a local antimigration factor against the pathogenesis of atherosclerosis and restenosis after angioplasty.
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Selected Abbreviations and Acronyms |
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Acknowledgments |
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The authors thank Makiko Ueda (Department of Pathology, Osaka City University Medical School), Nobuhiro Morisaki, and Masaki Kitahara (the Second Department of Internal Medicine, School of Medicine, Chiba University) for their helpful advice on the technique for migration assay. We also thank Atsumi Ohnishi and Tomoko Okuno for their technical assistance.
Received November 15, 1994; accepted June 6, 1995.
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 N. Nagaya, H. Okumura, M. Uematsu, W. Shimizu, F. Ono, M. Shirai, H. Mori, K. Miyatake, and K. Kangawa Repeated inhalation of adrenomedullin ameliorates pulmonary hypertension and survival in monocrotaline rats Am J Physiol Heart Circ Physiol, November 1, 2003; 285(5): H2125 - H2131. [Abstract] [Full Text] [PDF]
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 Y. Zhu and D. Y. Hui Apolipoprotein E Binding to Low Density Lipoprotein Receptor-related Protein-1 Inhibits Cell Migration via Activation of cAMP-dependent Protein Kinase A J. Biol. Chem., September 19, 2003; 278(38): 36257 - 36263. [Abstract] [Full Text] [PDF]
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N. Nagaya, K. Kangawa, M. Kanda, M. Uematsu, T. Horio, N. Fukuyama, J. Hino, M. Harada-Shiba, H. Okumura, Y. Tabata, et al. Hybrid Cell-Gene Therapy for Pulmonary Hypertension Based on Phagocytosing Action of Endothelial Progenitor Cells Circulation, August 19, 2003; 108(7): 889 - 895. [Abstract] [Full Text] [PDF]
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N. Fukai, M. Shichiri, N. Ozawa, M. Matsushita, and Y. Hirata Coexpression of Calcitonin Receptor-Like Receptor and Receptor Activity-Modifying Protein 2 or 3 Mediates the Antimigratory Effect of Adrenomedullin Endocrinology, February 1, 2003; 144(2): 447 - 453. [Abstract] [Full Text] [PDF]
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 M. Yamasaki, J. Kawai, T. Nakaoka, T. Ogita, A. Tojo, and T. Fujita Adrenomedullin Overexpression to Inhibit Cuff-Induced Arterial Intimal Formation Hypertension, February 1, 2003; 41(2): 302 - 307. [Abstract] [Full Text] [PDF]
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T. Tokudome, T. Horio, F. Yoshihara, S.-i. Suga, Y. Kawano, M. Kohno, and K. Kangawa Adrenomedullin Inhibits Doxorubicin-Induced Cultured Rat Cardiac Myocyte Apoptosis via a cAMP-Dependent Mechanism Endocrinology, September 1, 2002; 143(9): 3515 - 3521. [Abstract] [Full Text] [PDF]
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 J. Sun, X. Sui, J. A. Bradbury, D. C. Zeldin, M. S. Conte, and J. K. Liao Inhibition of Vascular Smooth Muscle Cell Migration by Cytochrome P450 Epoxygenase-Derived Eicosanoids Circ. Res., May 17, 2002; 90(9): 1020 - 1027. [Abstract] [Full Text] [PDF]
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H. Itoh, S. Yamamura, J. A. Ware, S. Zhuang, S. Mii, B. Liu, and K. C. Kent Differential effects of protein kinase C on human vascular smooth muscle cell proliferation and migration Am J Physiol Heart Circ Physiol, July 1, 2001; 281(1): H359 - H370. [Abstract] [Full Text] [PDF]
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T. Udono, K. Takahashi, M. Nakayama, A. Yoshinoya, K. Totsune, O. Murakami, Y. K. Durlu, M. Tamai, and S. Shibahara Induction of Adrenomedullin by Hypoxia in Cultured Retinal Pigment Epithelial Cells Invest. Ophthalmol. Vis. Sci., April 1, 2001; 42(5): 1080 - 1086. [Abstract] [Full Text]
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 P. D. Upton, J. Wharton, H. Coppock, N. Davie, X. Yang, M. H. Yacoub, M. A. Ghatei, J. M. Polak, S. R. Bloom, D. M. Smith, et al. Adrenomedullin Expression and Growth Inhibitory Effects in Distinct Pulmonary Artery Smooth Muscle Cell Subpopulations Am. J. Respir. Cell Mol. Biol., February 1, 2001; 24(2): 170 - 178. [Abstract] [Full Text]
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M. Sata, M. Kakoki, D. Nagata, H. Nishimatsu, E. Suzuki, T. Aoyagi, S. Sugiura, H. Kojima, T. Nagano, K. Kangawa, et al. Adrenomedullin and Nitric Oxide Inhibit Human Endothelial Cell Apoptosis via a Cyclic GMP-Independent Mechanism Hypertension, July 1, 2000; 36(1): 83 - 88. [Abstract] [Full Text] [PDF]
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T. Udono, K. Takahashi, M. Nakayama, O. Murakami, Y. K. Durlu, M. Tamai, and S. Shibahara Adrenomedullin in Cultured Human Retinal Pigment Epithelial Cells Invest. Ophthalmol. Vis. Sci., June 1, 2000; 41(7): 1962 - 1970. [Abstract] [Full Text]
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T. Horio, T. Nishikimi, F. Yoshihara, H. Matsuo, S. Takishita, and K. Kangawa Inhibitory Regulation of Hypertrophy by Endogenous Atrial Natriuretic Peptide in Cultured Cardiac Myocytes Hypertension, January 1, 2000; 35(1): 19 - 24. [Abstract] [Full Text] [PDF]
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S.-i. Hayashi, R. Morishita, H. Matsushita, H. Nakagami, Y. Taniyama, T. Nakamura, M. Aoki, K. Yamamoto, J. Higaki, and T. Ogihara Cyclic AMP Inhibited Proliferation of Human Aortic Vascular Smooth Muscle Cells, Accompanied by Induction of p53 and p21 Hypertension, January 1, 2000; 35(1): 237 - 243. [Abstract] [Full Text] [PDF]
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K. Shimizu, H. Tanaka, M. Sunamori, F. Marumo, and M. Shichiri Adrenomedullin Receptor Antagonism by Calcitonin Gene-Related Peptide(8-37) Inhibits Carotid Artery Neointimal Hyperplasia After Balloon Injury Circ. Res., December 3, 1999; 85(12): 1199 - 1205. [Abstract] [Full Text] [PDF]
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T. Tsuruda, J. Kato, K. Kitamura, M. Kawamoto, K. Kuwasako, T. Imamura, Y. Koiwaya, T. Tsuji, K. Kangawa, and T. Eto An autocrine or a paracrine role of adrenomedullin in modulating cardiac fibroblast growth Cardiovasc Res, September 1, 1999; 43(4): 958 - 967. [Abstract] [Full Text] [PDF]
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T. Horio, T. Nishikimi, F. Yoshihara, N. Nagaya, H. Matsuo, S. Takishita, and K. Kangawa Production and Secretion of Adrenomedullin in Cultured Rat Cardiac Myocytes and Nonmyocytes: Stimulation by Interleukin-1{beta} and Tumor Necrosis Factor-{alpha} Endocrinology, November 1, 1998; 139(11): 4576 - 4580. [Abstract] [Full Text] [PDF]
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H. Iwasaki, S. Eguchi, M. Shichiri, F. Marumo, and Y. Hirata Adrenomedullin as a Novel Growth-Promoting Factor for Cultured Vascular Smooth Muscle Cells: Role of Tyrosine Kinase-Mediated Mitogen-Activated Protein Kinase Activation Endocrinology, August 1, 1998; 139(8): 3432 - 3441. [Abstract] [Full Text] [PDF]
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M. Kohno, K. Yokokawa, K. Yasunari, M. Minami, H. Kano, T. Hanehira, and J. Yoshikawa Induction by Lysophosphatidylcholine, a Major Phospholipid Component of Atherogenic Lipoproteins, of Human Coronary Artery Smooth Muscle Cell Migration Circulation, July 28, 1998; 98(4): 353 - 359. [Abstract] [Full Text] [PDF]
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D. Palmer, K. Tsoi, and D. H. Maurice Synergistic Inhibition of Vascular Smooth Muscle Cell Migration by Phosphodiesterase 3 and Phosphodiesterase 4 Inhibitors Circ. Res., May 4, 1998; 82(8): 852 - 861. [Abstract] [Full Text] [PDF]
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Y. Isumi, H. Shoji, S. Sugo, T. Tochimoto, M. Yoshioka, K. Kangawa, H. Matsuo, and N. Minamino Regulation of Adrenomedullin Production in Rat Endothelial Cells Endocrinology, March 1, 1998; 139(3): 838 - 846. [Abstract] [Full Text] [PDF]
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K. Yasunari, M. Kohno, H. Kano, K. Yokokawa, M. Minami, and J. Yoshikawa Mechanisms of Action of Troglitazone in the Prevention of High Glucose-Induced Migration and Proliferation of Cultured Coronary Smooth Muscle Cells Circ. Res., December 19, 1997; 81(6): 953 - 962. [Abstract] [Full Text]
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K. Yasunari, M. Kohno, T. Hasuma, T. Horio, H. Kano, K. Yokokawa, M. Minami, and J. Yoshikawa Dopamine as a Novel Antimigration and Antiproliferative Factor of Vascular Smooth Muscle Cells Through Dopamine D1-Like Receptors Arterioscler. Thromb. Vasc. Biol., November 1, 1997; 17(11): 3164 - 3173. [Abstract] [Full Text]
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M. Kohno, K. Yokokawa, K. Yasunari, H. Kano, M. Minami, M. Ueda, and J. Yoshikawa Effect of Natriuretic Peptide Family on the Oxidized LDL–Induced Migration of Human Coronary Artery Smooth Muscle Cells Circ. Res., October 19, 1997; 81(4): 585 - 590. [Abstract] [Full Text]
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T. Sumimoto, T. Nishikimi, M. Mukai, K. Matsuzaki, E. Murakami, S. Takishita, A. Miyata, H. Matsuo, and K. Kangawa Plasma Adrenomedullin Concentrations and Cardiac and Arterial Hypertrophy in Hypertension Hypertension, September 1, 1997; 30(3): 741 - 745. [Abstract] [Full Text]
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M. Kohno, K. Yokokawa, H. Kano, K. Yasunari, M. Minami, T. Hanehira, and J. Yoshikawa Adrenomedullin Is a Potent Inhibitor of Angiotensin II–Induced Migration of Human Coronary Artery Smooth Muscle Cells Hypertension, June 1, 1997; 29(6): 1309 - 1313. [Abstract] [Full Text]
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