(Circulation. 2001;103:2955.)
© 2001 American Heart Association, Inc.
Basic Science Reports |
Functional Blockade of Platelet-Derived Growth Factor Receptor-ß but Not of Receptor-
Prevents Vascular Smooth Muscle Cell Accumulation in Fibrous Cap Lesions in Apolipoprotein E–Deficient Mice
Presented in part at the 72nd Scientific Sessions of the American Heart Association, Atlanta, Ga, November 7–10, 1999, and published in abstract form (Circulation. 1999;100[suppl I]:I-742).
From the Departments of Geriatric Medicine (H.S., M.Y., T.M., H.K., T.K.) and Molecular Genetics (T.S., N.T., S.N., S.-I.N.), Graduate School of Medicine, Kyoto University, Japan.
Correspondence to Masayuki Yokode, MD, Department of Geriatric Medicine, Kyoto University Graduate School of Medicine, 54 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan. E-mail yokode{at}kuhp.kyoto-u.ac.jp
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Abstract |
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Background—The vascular smooth muscle cell (VSMC) is the central cell component involved in the fibroproliferative response in atherogenesis. As the lesion advances, VSMCs migrate from the media into the subendothelial space, thereby forming fibrous plaque lesions. Platelet-derived growth factor (PDGF) has been known to be a potent chemoattractant and mitogen for SMCs, but the pathophysiological role of the 2 PDGF receptors, receptor-
(PDGFR-) and receptor-ß (PDGFR-ß) in atherogenesis
is poorly understood. To clarify this problem, we prepared
antagonistic rat monoclonal antibodies, APA5 and APB5,
against murine PDGFR- and PDGFR-ß,
respectively. Methods and Results—Apolipoprotein E–deficient mice were fed a high-fat diet containing 0.3% cholesterol from 6 weeks of age and subjected to injection with 1 mg/d IP of either antibody from 12 to 18 weeks every other day. In the mice injected with APB5, the aortic atherosclerotic lesion size and the number of intimal VSMCs were reduced by 67% and 80%, respectively, compared with the control mice injected with irrelevant rat IgG. In contrast, the mice that received APA5 showed only minimal reduction of lesion size, and a large number of VSMCs were observed in the intima. In the intima of advanced lesions, APB5 immunolabeled VSMCs, whereas APA5 could detect VSMCs mainly in the media.
Conclusions—These results indicate that PDGFR-ß plays a significant role in formation of fibrous atherosclerotic lesions and that regulation of the signal transduction through PDGFR-ß could affect atherogenesis in mice.
Key Words: platelet-derived factors • aorta • atherosclerosis • plaque • antibodies
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Introduction |
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TopAbstractIntroductionMethodsResultsDiscussionReferences |
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Atherosclerosis develops as a result of multiple inflammatory-fibroproliferative responses.1 The fatty streak lesions, filled mainly with macrophage-derived foam cells, proceed to intermediate lesions associated with vascular smooth muscle cells (VSMCs) migrating from the media. As pathological events continue, the intermediate lesions are converted into the further advanced lesions characterized by considerable accumulation of extracellular lipid, lipid-containing foam cells of macrophage and VSMC origin, and extracellular matrices.2
Recently, we reported that fatty streak formation was
prevented in apolipoprotein E (apoE)–deficient mice by administration
of an antagonistic monoclonal antibody (mAb) against murine
c-fms, the receptor for macrophage colony–stimulating
factor.3 This antibody had
little effect, however, on the advanced fibrous lesions associated with
a larger number of VSMCs in the intima. Among various growth factors,
platelet-derived growth factor (PDGF) released from
activated platelets, vascular endothelial
cells, VSMCs, and monocytes is an important mediator of VSMC
proliferation and migration.4
PDGF exists as a disulfide-linked dimer and is composed of 2 chains, A
and B.5 Two receptors for
PDGF, called and ß, have been
identified.6 Both receptors
are transmembrane glycoproteins with intrinsic tyrosine
kinase activity.7 Binding of
PDGF to the receptor induces receptor dimerization and activation of
the kinase activity. The -receptor (PDGFR-) binds both PGDF-A and
-B chains, whereas the ß-receptor (PDGFR-ß) binds only the PDGF-B
chain.8 9
The role of the PDGFR has been described in postinjury lesions by use of anti–PDGFR-ß antibody in the baboon.10 It has also been reported that both types of PDGF chains and their receptors could be detected in wound-healing processes of human coronary arteries after angioplasty.11 12 These results suggest that PDGF and its receptors are involved in the development of intimal lesions at sites of acute vascular injury.
Thus far, gene-targeting experiments have been attempted to
create knockout mice deficient for
PDGF-A,13
PDGF-B,14
PDGFR-,15 or
PDGFR-ß.16 Those mice,
however, died either at the embryonic stage or several days after
birth. This has made it difficult to study the significance of PDGF and
its receptors in atherogenesis in vivo and left unanswered the question
of whether PDGF could be involved in the natural course of
atherogenesis that occurs without provocation, such as mechanical
vascular injury.
To solve this problem, we administered 2 types of rat mAb,
APA5 and APB5, directed against murine PDGFR- and PDGFR-ß,
respectively, into 12-week-old apoE-deficient mice developing advanced
atherosclerotic lesions. We report that PDGFR- and PDGFR-ß have
distinct roles in progression and maintenance of the advanced
atherosclerotic regions associated with accumulation of
VSMCs.
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Methods |
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TopAbstractIntroductionMethodsResultsDiscussionReferences |
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Mice
The apoE-deficient mice were generous gifts from Dr Edward M. Rubin (University of California at Berkeley) and were hybrid 129ola x C57BL/6 mice.17 18 The mice were kept in a temperature-controlled facility on a 14 hours light/10 hours dark cycle with free access to food and water. After being weaned at 4 weeks of age, the mice were fed a normal chow diet (CMF, Oriental Yeast) until 6 weeks of age, when experiments were started.
Rat mAbs Directed Against Murine PDGFR-
and PDGFR-ß
APA5, a rat monoclonal anti–murine PDGFR-
antibody (IgG2a), was described
previously.19 Anti–murine
PDGFR-ß antibody, APB5, was prepared as follows. A cDNA fragment
corresponding to the extracellular domain of murine
PDGFR-ß20 was generated by
reverse transcription–polymerase chain reaction amplification of mRNA
prepared from the NIH3T3 cells. The amplified DNA fragment was inserted
into CD4Rg,21 from which the
CD4 gene had been removed. The DNA construct was transfected into the
COS-1 cell line. The PDGFR-ß/human IgG1 recombinant fusion protein
was purified from the culture supernatant of the transfected COS-1
cells. Spleen cells from a Wistar rat immunized and boosted with this
fusion protein were fused with X63.Ag8 cells, as described
previously.22
Detection of Autophosphorylated
PDGFR-ß
AC01 cells, a line of VSMCs established from
p53-deficient mice, were kindly provided by Dr Kazuhiro Ohmi (National
Children’s Research Center, Tokyo,
Japan).23 The synchronized
AC01 cells were preincubated with either APA5, APB5, or irrelevant
isotype-matched control rat IgG and were stimulated with 100 ng/mL of
PDGF-BB at 37°C for 5 minutes. The cells were lysed as described
previously,24 and the
PDGFR-ß was detected by immunoblotting with either
rabbit anti–murine PDGFR-ß polyclonal antibody, described
previously25 (generously
provided by Dr L.T. Williams, Chiron Corp) or mouse
anti-phosphotyrosine antibody 4G10 (Upstate Biotechnology). To detect
autophosphorylated PDGFR-ß after immunoprecipitation,
the cell lysates were incubated first with rabbit anti–human PDGFR-ß
antiserum (Upstate Biotechnology) and subjected to
immunoblotting with antibody
4G10.
Colony Assay
Colony assay was performed as described by Yan et
al.26 Briefly, bone marrow
cells from C57BL/6 mice were cultured with RPMI 1640 containing 15 U/mL
interleukin (IL)-3. The cells were given PDGF-BB (20 ng/mL) alone or
PDGF-BB plus either 20 µg/mL of APA5 or APB5. After 5-day culture,
the cells were counted and transferred to methylcellulose containing
IL-3 (200 U/mL) and erythropoietin (2 U/mL). After 2-day culture, the
number of colony formation units in culture was scored. In this assay,
a colony (>40 cells) is composed primarily of granulocytes and/or
macrophages.
Protocols for Feeding and Antibody
Administration
To study advanced and early stages of atherogenesis,
we designed 2 feeding protocols, protocol A and protocol B,
respectively. As illustrated in
Figure 1
, in protocol A, a total of 12 female apoE-deficient
mice were switched to a high-fat diet containing 20% fat and 0.3%
cholesterol (Oriental Yeast) at 6 weeks of age. In protocol
B, a total of 11 mice were kept on CMF for the entire experimental
period. In either protocol, 1 mg of APA5, APB5, or control rat IgG
(purified by 50% ammonium sulfate precipitation) was administered
intraperitoneally to each mouse on alternate days
from 12 to 18 weeks. At the end of the experimental period, mice were
killed by cervical dislocation and used for further analysis.
All experimental protocols were performed in accordance with the
guidelines of Kyoto University.
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Tissue Preparation and Histochemistry
The tissue preparation was conducted as described
previously.3 27
Briefly, the heart was removed from each mouse, snap-frozen in O.C.T.
Compound (Sakura Finetek USA, Inc), and sequentially cut into a total
of 36 cross sections (6 µm thick each) around the aortic sinus. Of
those sections, every third slice, ie, 12 samples per mouse, were
subjected to staining either with oil red O (Sigma Chemical); with a
biotin-labeled rat mAb, BM8, specific for mouse macrophage (BMA
Biochemicals AG); or with a mouse mAb, 1A4, against smooth muscle
-actin labeled with a horseradish peroxidase/EPOS system
(Dako).3 28 Each
section was counterstained with Meyer’s hematoxylin solution (Wako
Pure Chemical Industries). For analysis of the intimal VSMC
population, -actin–positive cells were counted in a total of 6
cross sections (every sixth section) from each mouse. To immunolabel
with APA5 and APB5, aortic sinus sections from apoE-deficient mice fed
a high-fat diet for 12 months were used. Each section was snap-frozen,
treated with either APA5 or APB5, incubated with peroxidase-conjugated
anti–rat IgG antibody, and reacted with TrueBlue Peroxidase Substrate
(Kirkegaard Perry Laboratories), which stained blue. The sections were
counterstained with nuclear fast red (Vector Laboratories), which
stained the nucleus red.
Image Analysis and Quantification of
Atherosclerotic Lesion
Atherosclerotic lesion size in each aortic section
was evaluated for oil red O staining by
Image-Pro Plus (Media Cybernetics). To estimate
the severity of the lesions, we calculated the "plaque ratio"
between the oil red O–stained area and the whole vessel area including
the lumen, intima, media, and adventitia as described by Nicoletti et
al.29 For each animal, 12
sections, ie, every third section, were examined, and the mean of the
fractional area was calculated and expressed as a
percentage.
Statistical Analysis
Data are expressed as mean±SD and were
analyzed by ANOVA with Abacus Statview software (version 4.5).
A value of P<0.05 was
considered statistically
significant.
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Results |
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TopAbstractIntroductionMethodsResultsDiscussionReferences |
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MAb APB5 Inhibited PDGFR-ß–Mediated Signal Pathway
We first examined whether mAb APB5 could block the PDGFR-ß–mediated signal pathway. When murine bone marrow cells were cultured with IL-3 and PDGF-BB, addition of APB5 suppressed proliferation of hematopoietic progenitors that are capable of forming a colony. In contrast, APA5, which has been shown to block the murine PDGFR-
pathway in vivo and in
vitro,19 had no suppressive
effect
(Table 1). We next studied whether the suppressive
effect of APB5 was due to blockade of the signal transduction pathway
through PDGFR-ß. As determined by immunoblot
analysis with anti-phosphotyrosine and anti–PDGFR-ß
antibodies, phosphorylation of PDGFR-ß in AC01 cells
was completely inhibited
(Figure 2, a, b, and c, lane 2). In contrast, APA5 or
irrelevant rat IgG had no inhibitory effect on
phosphorylation
(Figure 2, a and b, lanes 1, 3, and 4).
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Intraperitoneal
Administration of APB5 Prevented the Progression of Advanced
Atherosclerotic Lesions in ApoE-Deficient Mice
We asked whether administration of anti-PDGFR mAbs
could affect atherogenesis in apoE-deficient mice. For this purpose, we
designed 2 feeding protocols, protocols A and B, to examine the effect
of the antibodies on relatively advanced and early atherosclerotic
lesions, respectively
(Figure 1
). In protocol A, mice were fed a high-fat diet from
6 weeks of age so that the mice would develop fibroproliferative
lesions by 18 weeks of
age.3 18 The
animals were subjected to antibody treatment as described in Methods.
The mice that had been injected with irrelevant IgG developed advanced
atherosclerotic lesions in the aortic root as determined by staining
with oil red O
(Figure 3a). In contrast, the mice that received APB5 showed
marked reduction of aortic lesion size
(Figure 3b). The mice that had been given APA5 also showed a
tendency toward reduction of lesion size, whereas the extent of the
reduction was less than that seen in the mice injected with APB5
(Figure 3c). The plaque ratio in the mice injected with APB5
was as low as 33% of that in the control mice given irrelevant rat
IgG, ie, 5.02±2.93% (n=4) and 15.12±4.62% (n=4), respectively
(P=0.0049). Although
administration of APA5 slightly reduced the plaque ratio, by 28%,
compared with that of the control mice, ie, 10.84±3.82% (n=4), there
was no significant difference versus irrelevant rat IgG
(P=0.1501)
(Table 2). We next examined whether the antibodies could
have a similar effect on the relatively early lesions. For this
purpose, we designed protocol B, in which apoE-deficient mice were
maintained on CMF from 6 to 18 weeks of age so that they would develop
fatty streak lesions slowly in the
aorta.18 From 12 until 18
weeks of age, 1 mg of either APA5 or APB5 was administered on alternate
days. As determined with oil red O, the mice given either antibody had
tendency to show smaller lesions, whereas there was no significant
difference in plaque ratios: the plaque ratios in the mice given
APA5, APB5, and control rat IgG were 6.34±3.07% (n=4), 2.94±1.75%
(n=4), and 7.83±2.32% (n=3), respectively
(P=0.07).
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Administration of APB5 Markedly Prevented
Accumulation of VSMCs in Atheromatous Lesions in
ApoE-Deficient Mice
We further studied the effects of APB5 and APA5 on the
cell composition in the atheromatous lesions. First,
the aortic sections were stained with mAb 1A4 raised against -actin
to detect VSMCs. In the mice injected with irrelevant IgG or APA5, a
large number of the cells immunolabeled by 1A4 were clustering in the
intima and forming the fibrous cap structure overlying the
subendothelial
space28
(Figure 4, a, c, d, and f). We also counted the immunolabeled
cells in the aortic intima. The numbers of stained cells in the
examined sections from each mouse treated with irrelevant IgG and APA5
were 56.96±24.59 and 74.88±17.39, respectively
(Table 2). In striking contrast, the mice injected with APB5
presented a minimal number of such cells stained for -actin
in the intima, ie, 11.25±6.46 cells per mouse
(P<0.001 versus irrelevant IgG
or APA5)
(Figure 4, b and e)
(Table 2). In these mice, most of the cells stained with
anti–-actin were detected in the media. We next investigated the
distribution of macrophages in the aorta sections by
immunolabeling with the rat mAb BM8. The manner of distribution of
macrophages did not differ in the
subendothelial space in mice injected with either
irrelevant IgG, APB5, or APA5
(Figure 4, g, h, and i). Because it was suggested that
VSMCs in the intima were more sensitive to the action of APB5, we
investigated whether these cells were expressing PDGFR-ß by
immunohistochemical analysis, probing with APA5 and APB5. As
shown in
Figure 5, when the aortic lesions of 12-month-old
apoE-deficient mice that had been fed a high-fat diet since 6 weeks of
age were examined, the cells resident in the media and
atheromatous plaque were immunolabeled either with APA5
(Figure 5a) or with APB5
(Figure 5b). In striking contrast, the cells in the intima
were stained intensively with APB5 (indicated by arrow)
(Figure 5b) but minimally with APA5
(Figure 5a).
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Discussion |
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TopAbstractIntroductionMethodsResultsDiscussionReferences |
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In this article, we examine the roles of PDGFR-
and
PDGFR-ß in atherogenesis in apoE-deficient mice. We previously
reported that fatty streak formation was markedly suppressed in
12-week-old apoE-deficient mice by injection with AFS98, an
antagonistic rat mAb against murine c-fms, whereas this
antibody had minimal effect on the fibrous lesions with a large number
of VSMCs.3
On the basis of these observations, we attempted to seek the
mechanism by which the fibrous lesions are formed during this period.
It is still poorly understood by which mechanism VSMCs accumulate in
the intima during atherogenesis. Among numerous factors, PDGF has been
known to be involved both in migration and in proliferation of the
VSMCs. To test the hypothesis that PDGF is involved in formation of
such fibrous lesions, we prepared a novel rat mAb APB5 raised against
PDGFR-ß. The antagonistic effect of APB5 on the
PDGFR-ß–mediated pathway was confirmed by its effects on colony
formation of bone marrow cells
(Table 1) and on PDGFR-ß
autophosphorylation
(Figure 2). We previously demonstrated that APA5 blocks
specifically PDGFR- both in vivo and in
vitro.19 We confirmed
specific binding of APA5 and APB5 to PDGFR- and PDGFR-ß by
immunoblotting and immunoprecipitation (H.S. et al,
unpublished observations). Furthermore, injection with APA5 and APB5
had distinctive effects on kidney development in neonate mice (H.S. et
al, unpublished observations). These results demonstrate that APA5 and
APB5 did selectively block the signal pathways mediated by PDGFR-
and PDGFR-ß, respectively.
To examine the role of PDGF receptors in advanced and early
lesions of atheroma, we designed 2 feeding protocols of
apoE-deficient mice, protocols A and B, respectively
(Figure 1). In protocol A, we found that administration of
APB5 from 12 to 18 weeks of age was able to prevent the increase in the
atherosclerotic lesion size as determined by oil red O staining
(Figure 3). As assessed quantitatively, the lesion size of
the mice that had received APB5 was 33% of that of the control mice
injected with irrelevant IgG. APA5 had much less effect. These results
would imply that the PDGFR-ß–mediated signal transduction pathway
plays a significant role in atherogenesis from 12 to 18 weeks of age,
during which fibrous lesions are formed. In protocol B, we examined
whether APA5 and APB5 could exert similar action, in which foam cell
lesions are developed. APB5 showed a relatively weak tendency to reduce
the size of the foam cell–rich lesion, but we could detect no
significant difference from mice injected with APA5 or control IgG.
Although this could be due to the limited number of mice examined and
further study may be required, it was concluded that PDGFR-ß would be
more significantly involved in atherogenesis than PDGFR- in
apoE-deficient mice and that the blockade of PDGFR-ß–mediated signal
transduction could affect the development at least of advanced
atherosclerotic lesions.
In the present study, what was most noteworthy was
marked reduction of the density of intimal VSMCs in the mice injected
with APB5 as determined by immunostaining with
anti–-actin antibody 1A4
(Figure 4, b and e)
(Table 2). Because neither APA5 nor irrelevant rat IgG
caused such a change in the distribution pattern of VSMCs in the
arterial tissue
(Figure 4, a, c, d, and f), these results would suggest that
the preventive effect of APB5 on fibrous lesion formation is closely
correlated to blockade of the signal transduction system mediated by
PDGFR-ß. Interestingly, in the present study, the mice treated
with APA5 showed larger numbers of 1A4-labeled cells in the aortic
intima than did the irrelevant IgG–treated mice
(Table 2). Whether blockade of the PDGFR- pathway could
affect the change of intimal VSMC density must be investigated
further.
Thus far, several studies have proposed a potential role of PDGF and its receptors on the development of intimal hyperplasia at sites of acute vascular injury.10 30 31 32 Although these results suggest that PDGF and its receptors might play important roles in the development of intimal lesions at sites of acute vascular injury, it has been unclear whether PDGF could be involved in the natural course of atherogenesis that proceeds without provocation, such as mechanical vascular injury. Our data have demonstrated for the first time that PDGFR-ß is at least involved in the development of advanced atherosclerotic lesions. Because our present data indicate that inhibition of the PDGFR-ß pathway suppressed fibrous lesion formation markedly in vivo, it must be determined by which mechanism the PDGFR-ß pathway could be involved in the vessel wall. We are currently searching for the molecules that might be involved in signal transduction through either type of PDGFR in VSMCs.
In summary, we have developed a novel experimental system to investigate the behavior of VSMCs in the advanced lesion of atherosclerosis. In particular, we have shown that 2 types of PDGF receptor might have distinct regulatory roles in VSMCs and that the functional blockade of only PDGFR-ß was effective enough to change the size and cell composition of the relatively advanced lesions. Whether the regulation or management of the number of VSMCs could have a therapeutic role in subjects with atheromatous lesions that are prone to erosion or thrombosis must be investigated further.
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Acknowledgments |
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This research was supported by Ministry of Education, Science, Sports, and Culture of Japan research grants 04263104, 054040439, 0557052, 04304051, 08407026, and 9578; International Scientific Research Program grants 05044163, 07044254, and 09044293 from the Japanese Ministry of Education, Science, Sports, and Culture; a research grant for health sciences from the Japanese Ministry of Health and Welfare; grants 5A-2 and A8-1 for cardiovascular diseases from the Japanese Ministry of Health and Welfare; Grants-in-Aid for Scientific Research on Priority Areas 09281103 and 09281104; Grant-in-Aid for Creative Basic Research 09-NP-0601; the HMG-CoA Reductase Research Fund; the Japanese Foundation of Metabolism and Diseases; Takeda Medical Research Foundation; and grants from the Research Fellowships of Japan Society for the Promotion of Science for Young Scientists.
Received December 15, 2000; revision received February 27, 2001; accepted February 28, 2001.
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S. M. Schwartz, Z. S. Galis, M. E. Rosenfeld, and E. Falk Plaque Rupture in Humans and Mice Arterioscler Thromb Vasc Biol, April 1, 2007; 27(4): 705 - 713. [Abstract] [Full Text] [PDF]
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J.-H. Wu, R. Goswami, X. Cai, S. T. Exum, X. Huang, L. Zhang, L. Brian, R. T. Premont, K. Peppel, and N. J. Freedman Regulation of the Platelet-derived Growth Factor Receptor-beta by G Protein-coupled Receptor Kinase-5 in Vascular Smooth Muscle Cells Involves the Phosphatase Shp2 J. Biol. Chem., December 8, 2006; 281(49): 37758 - 37772. [Abstract] [Full Text] [PDF]
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D. L. Tharp, B. R. Wamhoff, J. R. Turk, and D. K. Bowles Upregulation of intermediate-conductance Ca2+-activated K+ channel (IKCa1) mediates phenotypic modulation of coronary smooth muscle Am J Physiol Heart Circ Physiol, November 1, 2006; 291(5): H2493 - H2503. [Abstract] [Full Text] [PDF]
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T. Raj, P. Kanellakis, G. Pomilio, G. Jennings, A. Bobik, and A. Agrotis Inhibition of Fibroblast Growth Factor Receptor Signaling Attenuates Atherosclerosis in Apolipoprotein E-Deficient Mice Arterioscler Thromb Vasc Biol, August 1, 2006; 26(8): 1845 - 1851. [Abstract] [Full Text] [PDF]
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T. Takahashi, H. Abe, H. Arai, T. Matsubara, K. Nagai, M. Matsuura, N. Iehara, M. Yokode, S. Nishikawa, T. Kita, et al. Activation of STAT3/Smad1 Is a Key Signaling Pathway for Progression to Glomerulosclerosis in Experimental Glomerulonephritis J. Biol. Chem., February 25, 2005; 280(8): 7100 - 7106. [Abstract] [Full Text] [PDF]
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K. Peppel, L. Zhang, E. S. Orman, P.-O. Hagen, A. Amalfitano, L. Brian, and N. J. Freedman Activation of vascular smooth muscle cells by TNF and PDGF: overlapping and complementary signal transduction mechanisms Cardiovasc Res, February 15, 2005; 65(3): 674 - 682. [Abstract] [Full Text] [PDF]
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M. Campbell and E. R. Trimble Modification of PI3K- and MAPK-Dependent Chemotaxis in Aortic Vascular Smooth Muscle Cells by Protein Kinase C{beta}II Circ. Res., February 4, 2005; 96(2): 197 - 206. [Abstract] [Full Text] [PDF]
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C. C.M. Appeldoorn, A. Bonnefoy, B. C.H. Lutters, K. Daenens, T. J.C. van Berkel, M. F. Hoylaerts, and E. A.L. Biessen Gallic Acid Antagonizes P-Selectin-Mediated Platelet-Leukocyte Interactions: Implications for the French Paradox Circulation, January 4, 2005; 111(1): 106 - 112. [Abstract] [Full Text] [PDF]
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K. L. Hildreth, J.-H. Wu, L. S. Barak, S. T. Exum, L. K. Kim, K. Peppel, and N. J. Freedman Phosphorylation of the Platelet-derived Growth Factor Receptor-{beta} by G Protein-coupled Receptor Kinase-2 Reduces Receptor Signaling and Interaction with the Na+/H+ Exchanger Regulatory Factor J. Biol. Chem., October 1, 2004; 279(40): 41775 - 41782. [Abstract] [Full Text] [PDF]
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F. Dandre and G. K. Owens Platelet-derived growth factor-BB and Ets-1 transcription factor negatively regulate transcription of multiple smooth muscle cell differentiation marker genes Am J Physiol Heart Circ Physiol, June 1, 2004; 286(6): H2042 - H2051. [Abstract] [Full Text] [PDF]
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T. Kadowaki and N. Kubota Protective Role of Imatinib in Atherosclerosis Arterioscler Thromb Vasc Biol, May 1, 2004; 24(5): 801 - 803. [Full Text]
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M. Lassila, T. J. Allen, Z. Cao, V. Thallas, K. A. Jandeleit-Dahm, R. Candido, and M. E. Cooper Imatinib Attenuates Diabetes-Associated Atherosclerosis Arterioscler Thromb Vasc Biol, May 1, 2004; 24(5): 935 - 942. [Abstract] [Full Text]
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H. Sano, Y. Ueda, N. Takakura, G. Takemura, T. Doi, H. Kataoka, T. Murayama, Y. Xu, T. Sudo, S. Nishikawa, et al. Blockade of Platelet-Derived Growth Factor Receptor-{beta} Pathway Induces Apoptosis of Vascular Endothelial Cells and Disrupts Glomerular Capillary Formation in Neonatal Mice Am. J. Pathol., July 1, 2002; 161(1): 135 - 143. [Abstract] [Full Text] [PDF]
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