© 2001 American Heart Association, Inc.
Atherosclerosis and Lipoproteins |
Oxidized LDL Regulates Vascular Endothelial Growth Factor Expression in Human Macrophages and Endothelial Cells Through Activation of Peroxisome Proliferator–Activated Receptor-
From the Department of Medicine and Clinical Science, Kyoto University Graduate School of Medicine, Kyoto, Japan.
Correspondence to Hiroshi Itoh, MD, PhD, Department of Medicine and Clinical Science, Kyoto University Graduate School of Medicine, 54 Shogoin Kawahara-cho, Sakyo-ku, Kyoto 606-8507, Japan. E-mail hiito{at}kuhp.kyoto-u.ac.jp
 |
Abstract |
|---|
 Top Abstract IntroductionMethodsResultsDiscussionReferences |
|---|
Abstract—Vascular endothelial growth factor (VEGF) has been recognized as an angiogenic factor that induces endothelial proliferation and vascular permeability. Recent studies have also suggested that VEGF can promote macrophage migration, which is critical for atherosclerosis. We have reported that VEGF is remarkably expressed in activated macrophages, endothelial cells, and smooth muscle cells within human coronary atherosclerotic lesions, and we have proposed the significance of VEGF in the progression of atherosclerosis. To clarify the mechanism of VEGF expression in atherosclerotic lesions, we examined the regulation of VEGF expression by oxidized low density lipoprotein (Ox-LDL), which is abundant in atherosclerotic arterial walls. A recent report has revealed that peroxisome proliferator–activated receptor-
(PPAR) is expressed not only in adipocytes but also in
monocytes/macrophages and has suggested that PPAR may have a
role in the differentiation of monocytes/macrophages.
Furthermore, 9- and
13-hydroxy-(S)-10,12-octadecadienoic
acid (9- and 13-HODE, respectively), the components of Ox-LDL, may be
PPAR ligands. Therefore, we investigated the involvement of PPAR
in the regulation of VEGF by Ox-LDL. PPAR expression was detected in
human monocyte/macrophage cell lines, human acute monocytic
leukemia (THP-1) cells, and human coronary artery
endothelial cells (HCAECs). Ox-LDL (10 to 50 µg/mL)
upregulated VEGF secretion from THP-1 dose-dependently. VEGF mRNA
expression in HCAECs was also upregulated by Ox-LDL. The mRNA
expression of VEGF in THP-1 cells and HCAECs was also augmented by
PPAR activators, troglitazone (TRO), and
15-deoxy-
12,14-prostaglandin
J2 (PGJ2). In contrast, VEGF expression in
another monocyte/macrophage cell line, human histiocytic
lymphoma cells (U937), which lacks PPAR expression, was not
augmented by TRO or PGJ2. We established the U937 cell line, which
permanently expresses PPAR (U937T). TRO and Ox-LDL augmented VEGF
expression in U937T. In addition, VEGF production by THP-1
cells was significantly increased by exposure to 9-HODE and 13-HODE. In
conclusion, Ox-LDL upregulates VEGF expression in macrophages
and endothelial cells, at least in part, through the
activation of
PPAR.
Key Words: vascular endothelial growth factors • atherosclerosis • macrophages • endothelial cells • peroxisome proliferator–activated receptor-
|
Introduction |
|---|
|
TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
Vascular endothelial growth factor (VEGF) has been recognized as an angiogenic factor that induces endothelial proliferation and vascular permeability.1 It may play a role in tumor growth, wound healing, age-related macular degeneration, rheumatoid arthritis, diabetic retinopathy, and collateral formation in ischemic tissue. However, recent studies have demonstrated that the gene for the VEGF receptor flt-1 is also expressed in human monocytes and that the VEGF-induced chemotactic response of human monocytes is mediated via this receptor.2 Clauss et al3 have also documented that VEGF induces monocyte activation and migration. These data strongly suggest that VEGF plays a role in the chemotaxis of monocytes/macrophages, which is crucial in inflammatory reactions and in wound-repair processes. We and others have reported that VEGF is remarkably expressed in activated macrophages, endothelial cells, and smooth muscle cells within human coronary atherosclerotic lesions, and we have proposed the significance of VEGF in the progression of atherosclerosis.4 5 6 7
Oxidized LDL (Ox-LDL) has been demonstrated to be a key molecule in the atherosclerotic process.8 9 It has been reported that Ox-LDL, which abundantly exists in atherosclerotic arterial walls, plays an important role in endothelial functional alterations, the recruitment and retention of monocyte/macrophages, and foam cell formation.10 11 12 It has been also revealed that Ox-LDL modulates the secretion of cytokines or growth factors from endothelial cells and macrophages.13 14
In this context, we hypothesized that Ox-LDL might play a role in the expression of VEGF in injured endothelial cells or accumulated macrophages in atherosclerotic lesions. In the present study, to clarify the mechanism of VEGF expression in atherosclerotic lesions, we examined the regulation of VEGF expression in endothelial cells and macrophages by Ox-LDL.
Peroxisome proliferator–activated receptor-
(PPAR) is a member of the nuclear receptor superfamily of
ligand-activated transcription
factors.15 PPAR and
retinoid X receptor contain the heterodimer to bind regulatory elements
in the promoter region of a number of adipocyte-specific genes and to
stimulate transcription in response to PPAR-specific and retinoid X
receptor–specific
ligands.16 17
PPAR mRNA is most highly expressed in adipose tissue and plays an
important role in regulating adipocyte differentiation and glucose
metabolism.18 A
new class of antidiabetic agents, thiazolidinediones, have been
developed and have been demonstrated to improve insulin resistance by
binding and activating
PPAR.19 20 21
Recently, Tontonoz et
al22 have reported that
PPAR is expressed not only in adipocytes but also in
monocytes/macrophages, and they have suggested that PPAR may
have a role in the differentiation of monocytes/macrophages.
Moreover, they have identified 2 of the major oxidized lipid
components of Ox-LDL,
9-hydroxy-(S)-10,12-octadecadienoic
acid (9-HODE) and
13-hydroxy-(S)-10,12-octadecadienoic
acid (13-HODE), as endogenous activators and
ligands of PPAR.23 PPAR
expression in human endothelial cells has also been
revealed.24
Thus, we sought to determine whether PPAR might work
through the pathway of regulation of VEGF expression in
endothelial cells and macrophages stimulated by
Ox-LDL. In the present study, we further investigate the regulation
of VEGF expression by Ox-LDL in macrophages and
endothelial cells via the possible activation of
PPAR activators.
|
Methods |
|---|
|
TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
Agents
Troglitazone (TRO) was obtained from Sankyo; 15-deoxy-
12,14-prostaglandin
J2 (PGJ2), prostaglandin
F2
(PGF2), 9-HODE,
and 13-HODE were purchased from Cayman. LDL (Sigma Chemical Co) was
dialyzed against 3 changes of PBS to remove EDTA. Then LDL was oxidized
at a concentration of 500 µg/mL by exposure to 10 µmol/L
CuSO4 for 24 hours at room temperature, followed
by dialysis at 4°C for 24 hours against 3 changes of
PBS.12 14 25
We designated LDL incubated with CuSO4 for 5
hours as minimum modified LDL (mm-LDL) and LDL incubated for 24 hours
as Ox-LDL. The extent of lipid peroxidation was estimated by
electrophoresis in agarose gel.
Cell Culture
Human monocytic leukemia (THP-1) cells and human
monoblastic leukemia (U937) cells were obtained from the American
Tissue Culture Collection and cultured in RPMI 1640 medium (GIBCO-BRL)
with 10% FBS at 37°C in 5% CO2. Cells were
cultured in serum-free medium just before the experiments. The U937
cell line permanently expressing PPAR was established as follows:
PPAR1 expression vector,
pCMX-mPPAR,26 possessing
cytomegalovirus enhancer and mouse full-length PPAR1
cDNA27 (1.4 kb, GenBank No.
U10374), which was kindly donated by Prof K. Umesono (Kyoto University
Graduate School of Medicine), and 10 mg neomycin resistance gene
plasmid expression vector
(pCXN-Neo)28 were
cotransfected to U937. Electroporation was performed by using a Bio-Rad
Gene Pulser set at 220 V and 960 mF. Neomycin (GIBCO-BRL) was added to
a final concentration of 1 g/mL. Individual clones were isolated by
seeding transfected cells at a limiting dilution in 96-well plates in
the medium containing 20% FBS. We designated this U937 cell line
expressing PPAR as U937T cells. Human coronary artery
endothelial cells (HCAECs) were purchased from
Clonetics, and they were cultured in EGM-2MV (endothelial cell
growth medium; Clonetics) with 5% FCS and changed to serum-free medium
just before the experiments.
RNA Extraction, Northern Blot
Analysis, and RT-PCR
Total cellular RNA was isolated from HCAECs and THP-1
cells by using TRIzol reagents (GIBCO-BRL). RNA (20 µg per lane) was
electrophoresed on 1% agarose/6% formaldehyde gels and transferred to
nylon membranes (Biodyne, Pall BioSupport). Hybridization was performed
at 42°C for 20 hours in 50% formamide, 5x SSC, 0.05 mol/L phosphate
buffer, 5x Denhardt’s solution, 0.1% SDS, 0.1 mg/mL denatured salmon
sperm DNA, and 1x109 cpm/mL
32P-labeled cDNA probes. A 320-bp
PstI-EcoRI
fragment (nucleotides 484 to 501) of rat VEGF189 cDNA was
used as a probe. After hybridization, the filters were washed in 1x
SSC/0.1% SDS for 10 minutes at room temperature, for 30 minutes at
55°C, and then in 0.1x SSC/0.1% SDS for 30 minutes at 55°C.
Filters were autoradiographed, and the resulting bands were quantified
by the BAS 2000 system (Fuji Photo Film Co, Ltd). Reverse transcriptase
(RT)–polymerase chain reaction (PCR) for VEGF mRNA was performed as
follows5 : cDNA was synthesized
from 5 µg of total RNA primed with oligo-dT by use of RT (Super
Script II) at 42°C for 1 hour. cDNA (0.5 µg) was subjected to PCR
with 50 fmol of primers. Primer sequences were
5'-GGACATCTTCCAGGAGTA-3' (human VEGF165 cDNA
nucleotides 232 to 250) and 5'-TGCAACGCGAGTCTGTGT-3'
(nucleotides 555 to 573) for detecting human VEGF mRNA.
Amplification was carried out for 30 cycles with 30 seconds of
incubation at 94°C, 30 seconds at 55°C, and 1 minute at 72°C. A
10-µL aliquot of each RT-PCR reaction mixture was electrophoresed on
a 1.5% agarose gel.
VEGF Protein Quantification
After incubation of the THP-1, U293, and U293T under
the indicated conditions, the conditioned medium was collected and
centrifuged at 100g for
5 minutes, and the supernatant was used for the measurement of VEGF
protein with a Quantikine Human VEGF ELISA kit (R&D Systems, Inc). The
sensitivity of the assay was 1 pg/mL. All samples were measured in
duplicate.
Immunohistochemical Staining
HCAECs were cultured by use of flexiPERM (Heraeus
Instruments), and before staining, they were
permeabilized by Triton-X (Nakarai Tesque). For the
identification of VEGF, a rabbit polyclonal antibody that was directed
against the 20 amino terminal residues of human VEGF (Santa Cruz
Biotechnology) was used. Immunohistochemical staining was performed by
the LSAB (labelled streptavidin-biotin) method as
described.5
Western Blotting
Nuclear extracts of THP-1, U937, and U937T cells were
obtained as described.29
Samples were applied to 10% SDS gels and transferred to nitrocellulose
membranes (Bio-Rad) by using semidry blotting. Anti-PPAR polyclonal
antibody was a kind gift from Dr Heidi Camp (Pfizer Inc, Ann Arbor,
Mich).29 Membranes were
incubated with the antibody at a dilution of 1:1000 for 2 hours. After
they were washed, the membranes were stained with horseradish
peroxidase-conjugated rabbit anti-goat antibody. Antigen detection was
performed with a CSL kit (Amersham-Pharmacia).
Statistical Analysis
All results were expressed as mean±SD. Statistical
analyses of the data were performed by ANOVA. A value of
P<0.05 was considered
significant. The experiments presented were the
representatives of at least 2 separate
experiments.
|
Results |
|---|
|
TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
Ox-LDL Increases VEGF Expression in Endothelial Cells and Monocytes/Macrophages
We have previously reported that human endothelial cells express VEGF in atherosclerotic lesions.5 We examined whether Ox-LDL stimulates VEGF expression in HCAECs. As shown in Figure 1A
, Ox-LDL (10 to 50 µg/mL) increased VEGF mRNA
expression in HCAECs in a time-dependent manner. When HCAECs were
exposed to 10 to 50 µg/mL Ox-LDL for 6 hours, the VEGF mRNA level
increased in a dose-dependent manner
(Figure 1B). In contrast, native LDL at the same
concentration had no effect on VEGF mRNA levels. We also used
immunohistochemical techniques to examine whether VEGF was present
in cultured HCAECs. As shown in
Figure 1C, strong positive staining for VEGF was observed in
HCAECs cultured with 100 µg/mL Ox-LDL for 12 hours. Native LDL had no
effect on VEGF staining. No evidence of a toxic effect of Ox-LDL was
observed during the incubation with HCAECs, as assessed by morphology
and trypan blue exclusion.
|
THP-1 cells were exposed to Ox-LDL at concentrations between 10 and 50 µg/mL for 24 hours. VEGF levels were determined in culture medium by ELISA. VEGF concentration was increased significantly by exposure to Ox-LDL (for VEGF concentration, vehicle was 548.7±28.4 pg/mL, 10 µg/mL Ox-LDL was 742.0±26.7 pg/mL [P<0.05 versus vehicle], and 50 µg/mL Ox-LDL was 1078.9±100.6 pg/mL [P<0.05 versus vehicle]). Native LDL had no effect on VEGF protein levels (599.1±3.3 pg/mL). No evidence of a toxic effect of Ox-LDL was observed during the incubation with THP-1 cells.
PPAR Activators Increase VEGF
Expression in Endothelial Cells and
Monocytes/Macrophages
HCAECs were exposed to TRO
(10-7 to
10-5 mol/L). As
shown in
Figure 2A, VEGF mRNA expression was upregulated by TRO in a
dose-dependent manner (percent over the control,
10-7 mol/L TRO
159%, 10-6
mol/L TRO 193%, and
10-5 mol/L TRO
250%) The mRNA expression of VEGF in THP-1 was also dose-dependently
augmented by TRO and PGJ2 at
10-5 to
10-7 mol/L
(percent over the control,
10-7 mol/L TRO
134%, 10-6
mol/L TRO 146%,
10-5 mol/L TRO
170%, 10-7
mol/L PGJ2 122%,
10-6 mol/L PGJ2
143%, 10-5
mol/L PGJ2 168%, and 12-0-tetradecanoylphorbol-13-acetate
[TPA] 203%;
Figure 2B). In contrast, VEGF expression in U937 was not
augmented by TRO or PGJ2 (data not shown). As shown in
Figure 2C, RT-PCR analysis also confirmed that TRO
(10-5 mol/L) as
well as lysophosphatidylcholine (LPC,
10-5 mol/L),
which is one of the major components of Ox-LDL, increased VEGF mRNA
expression. The augmentation of VEGF mRNA expression by TRO or LPC in
THP-1 cells was not observed in the presence of actinomycin D, the
inhibitor of transcription.
|
PPAR Activators Increase VEGF
Protein Levels in Culture Medium
To examine whether PPAR activators
stimulate VEGF production by THP-1 cells, VEGF levels were
determined by ELISA in culture medium from THP-1 cells incubated with
or without PPAR activators for 24 hours.
Consistent with the results of alterations of mRNA expression,
VEGF production by THP-1 cells was increased by exposure to TRO
(5 and 10 µmol/L) in a time-dependent manner
(Figure 3A). TRO and PGJ2 stimulated VEGF production
by THP-1 cells in a concentration-dependent manner
(Figure 3B and 3C). In the culture medium collected after
treatment with 10 µmol/L TRO, the VEGF concentration was 3.9-fold
higher than that of control
(Figure 3B). We further examined the effect of
PGF2, an agent known to inhibit PPAR
activation,30 on TRO-induced
secretion of VEGF from THP-1. The addition of
PGF2 (200 nmol/L) significantly decreased the
effect of TRO on VEGF secretion
(Figure 3D).
|
Ox-LDL Upregulates VEGF Expression Partly
Through PPAR Activation
To further characterize the role of PPAR in
Ox-LDL–regulated VEGF expression in monocytes/macrophages, we
established the permanent cell line of U937 expressing PPAR (U937T
cells). As shown in
Figure 4A, in U937T cells, TRO treatment resulted in a
marked upregulation of VEGF secretion similar to that in THP-1 cells,
whereas it had no effect on VEGF secretion from U937 cells. As shown in
Figure 4B, PPAR expression in THP-1 cells was barely
detected by Western blot, but after TRO or TPA stimulation, we could
detect the expression of PPAR in THP-1 cells. U937T cells showed
significant PPAR expression, whereas U937 cells did not.
Figure 5 shows the responsiveness of THP-1, U937, and U937T
cells to Ox-LDL. VEGF secretion from these cell lines was significantly
upregulated by Ox-LDL stimulation with different magnitudes. VEGF
secretion from U937T cells was increased to 190% and 212% by 10 and
50 µg/mL Ox-LDL stimulation, respectively, whereas the secretion from
THP-1 cells was increased to 135% and 197% by Ox-LDL at 10 and 50
µg/mL, respectively. The increase of VEGF secretion from U937 cells
by Ox-LDL (50 µg/mL) was only 125% of the control value. These
differences suggest that PPAR plays some role in the VEGF expression
in THP-1 cells induced by Ox-LDL.
|
|
9-HODE and 13-HODE Regulate VEGF Secretion From
Human Macrophage Cell Lines
Two of the major oxidized lipid components of Ox-LDL,
9-HODE and 13-HODE, were reported to be endogenous
activators and ligands of PPAR. Thus, we investigated
whether 9-HODE and 13-HODE could stimulate VEGF production by
human macrophage cell lines. As shown in
Figure 6, VEGF production by THP-1 cells was
increased by exposure to 9-HODE and 13-HODE (0.1 to 5 µg/mL) for 24
hours, but VEGF production by U937 cells, in which we could not
detect PPAR mRNA, was not increased by 9-HODE or 13-HODE. The result
further supports the possibility that Ox-LDL induces VEGF expression
through PPAR activation. In addition, LPC also stimulated VEGF
production by U937 cells.
|
|
Discussion |
|---|
|
TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
We and others have previously demonstrated the expression of VEGF protein and mRNA in the progression of human coronary atherosclerosis.4 5 6 7 We have reported that VEGF is remarkably expressed in activated macrophages, endothelial cells, and smooth muscle cells within human coronary atherosclerotic lesions and proposed the significance of VEGF in the progression of atherosclerosis. Because Ox-LDL is abundant in atherosclerotic arterial walls and is critically involved in atherogenesis, we hypothesized that Ox-LDL might have a role in the expression of VEGF in atherosclerotic lesions. Ramos et al6 have previously reported the induction of VEGF expression in a mouse macrophage-like cell line (RAW 264 cells) in response to Ox-LDL. The present study was compatible with their result and further revealed that Ox-LDL–induced upregulation of VEGF occurred not only in macrophages but also in endothelial cells.
The significance of PPAR in
atherosclerosis is beginning to be elucidated. It is
reported that PPAR activators inhibit
neointimal formation after balloon injury of the rat
carotid artery.31 Recently,
many lines of evidence have elucidated that PPAR regulates the
expression of several genes important not only for lipid
metabolism but also for
atherogenesis.15 31 32
We have reported that PPAR activators regulate monocyte
migration.33 We have also
revealed that PPAR activators modulate
endothelial cell growth and secretion of endothelin-1
and C-type natriuretic
peptide.34 In the present
study, we showed that PPAR ligands, TRO and PGJ2, upregulated VEGF
expression in HCAECs and THP-1 cells, which express PPAR.
Furthermore, the effects of TRO on VEGF mRNA expression in THP-1
cells were not seen in the presence of actinomycin D. This result
suggests that the increase of the VEGF mRNA level by TRO was at the
transcriptional level. In contrast, they had no effect on VEGF
expression in wild-type U937 cells, which lack the expression of
PPAR. We have further demonstrated that the U937 cell line, which
stably expresses PPAR, restores the responsiveness of VEGF
expression to TRO . In addition, PGF2, which
has been shown to inhibit phosphorylation of PPAR
and exert inhibition on PPAR
activation,30 significantly
suppressed the effect of TRO on VEGF secretion from THP-1 cells. Taken
together, it is likely that PPAR activation can cause the induction
of VEGF expression.
However, it remains unclear how PPAR upregulates VEGF
gene expression. We searched the promoter region of the VEGF gene but
could not find the direct repeat sequence of the PPAR responsive
element. Recently, many coactivators, such as CBP/p300 and
PPAR binding protein, which interact with multiple ligand-bound nuclear
receptors and facilitate their transactivation, have been
isolated.35 36 The
interaction with PPAR and these coactivators may have
influences on the effect of other transcriptional factors, which
require these coactivators. Elucidation of the mechanism of
PPAR-dependent regulation of VEGF gene expression requires further
investigation.
PPAR is activated by the lipid components of
Ox-LDL (9-HODE and 13-HODE) and
PGJ2.22 23 9-HODE
and 13-HODE have been shown to promote the differentiation of human
monocytes to macrophages and to facilitate Ox-LDL uptake
through transcriptional induction of the scavenger receptor CD36. Thus,
we hypothesized that VEGF expression in endothelial
cells and macrophages stimulated by Ox-LDL is regulated through
the activation of PPAR. There are only a few reports concerning the
significance of Ox-LDL and its components, 9-HODE and 13-HODE, as
natural ligands of PPAR. Chen et
al37 have reported that the
PPAR agonists enhance endothelial cell adhesiveness
by increasing intercellular adhesion molecule-1 (ICAM-1) expression.
They have shown that ICAM-1 expression is upregulated by the PPAR
ligand TRO and also by Ox-LDL as well as 13-HODE and have suggested
that the PPAR signaling pathway might contribute to the regulation
of ICAM-1 and atherogenicity of Ox-LDL. In the present study, we
have demonstrated the upregulation of VEGF expression in
macrophages stimulated by 9-HODE and 13-HODE. Moreover, we have
observed that U937T cells, which permanently express PPAR, show
responsiveness to Ox-LDL stimulation similar to that in THP-1 cells,
which express PPAR. These data strongly suggest that VEGF expression
in endothelial cells and macrophages stimulated
by Ox-LDL is regulated, at least in part, through the activation of
PPAR.
Ramos et al6 have
shown that LPC also upregulates VEGF expression in RAW 264 cells. We
have observed that LPC also induces VEGF expression in
HCAECs.38 As shown in
Figure 5
, Ox-LDL stimulated VEGF secretion in U937 cells
with less magnitude. Inasmuch as we observed significant stimulation by
LPC of VEGF expression in U937 cells
(Figure 2C and 6), LPC is also involved in Ox-LDL–regulated
VEGF expression.
The exact role of VEGF in atherosclerotic lesions remains to
be determined. It has been reported that the in vivo introduction of
human VEGF165 cDNA into rabbit carotid arteries, by use of the
hemagglutinating virus of Japan–liposome method, induces prominent
angiomatoid proliferation of endothelial cells and
thickening of the intima due to fibromuscular
hyperplasia.39 Angiogenesis
in atheromatous plaques is suggested to be an important
process of atherogenesis and plaque
rapture.40 41 Thus,
VEGF produced in accumulating macrophages in
atheromatous plaque could have a proatherogenic effect.
On the other hand, VEGF can also play a role in protecting
endothelial cells from injury in the early phase of
atherosclerosis.42
Upregulation of VEGF expression by Ox-LDL, the key molecule of
atherosclerosis, through the PPAR activation
observed in the present study can be relevant in the progression of
atherosclerosis.
In conclusion, we have shown that Ox-LDL regulates VEGF
expression in human atherosclerotic lesions through, at least in part,
ligand activation of PPAR. These findings suggest that the PPAR
signaling pathway may play a role in atherogenesis induced by
Ox-LDL.
|
Acknowledgments |
|---|
This work was supported in part by research grants from the Japanese Ministry of Education, Science, and Culture; the Japanese Society for the promotion of Science "Research for the Future" program (JSPS-RFTF 96100204 and JSPS-RFTF 98L00801); and the smoking research foundation. We appreciate technical advice concerning the preparation of Ox-LDL from Dr Kiyotaka Kugiyama (Kumamoto University, Japan). We thank Prof Alan R. Saltiel (University of Michigan, Pfizer Inc), Dr Heidi Camp (Pfizer Inc), and Liyun Ding (Pfizer Inc) for their technical advice and gift of PPAR
antibody.
We thank Ayumi Sone, Akane Nonoguchi, and Yukiho Takada for their
excellent secretarial
work. Received July 3, 2000; accepted December 12, 2000.
|
References |
|---|
|
TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
1. Ferrara N, Houck K, Jakeman L, Leung DW. Molecular and biological properties of the vascular endothelial growth factor family of proteins. Endocr Rev. 1992;13:18–32.
2.
Shen H, Clauss M,
Ryan J, Schmidt AM, Tijburg P, Borden L, Connolly D, Stern D, Kao
J. Characterization of vascular permeability factor/vascular
endothelial growth factor receptors on mononuclear
phagocytes. Blood. 1993;81:2767–2773.
3.
Clauss M, Gerlach M,
Gerlach H, Brett J, Wang F, Familletti PC, Pan YC, Olander JV, Connolly
DT, Stern D. Vascular permeability factor: a tumor-derived polypeptide
that induces endothelial cell and monocyte procoagulant
activity, and promotes monocyte migration.
J Exp Med. 1990;172:1535–1545.
4. Couffinhal T, Kearney M, Witzenbichler B, Chen D, Murohara T, Losordo DW, Symes J, Isner JM. Vascular endothelial growth factor/vascular permeability factor (VEGF/VPF) in normal and atherosclerotic human arteries. Am J Pathol. 1997;150:1673–1685.[Abstract]
5.
Inoue M, Itoh H,
Ueda M, Naruko T, Kojima A, Komatsu R, Doi K, Ogawa Y, Tamura N, Takaya
K, et al. Vascular endothelial growth factor (VEGF)
expression in human coronary atherosclerotic lesions: possible
pathophysiological significance of VEGF in
progression of atherosclerosis.
Circulation. 1998;98:2108–2116.
6.
Ramos MA, Kuzuya M,
Esaki T, Miura S, Satake S, Asai T, Kanda S, Hayashi T, Iguchi A.
Induction of macrophage VEGF in response to oxidized LDL and
VEGF accumulation in human atherosclerotic lesions.
Arterioscler Thromb Vasc Biol. 1998;18:1188–1196.
7.
Chen YX, Nakashima
Y, Tanaka K, Shiraishi S, Nakagawa K, Sueishi K. Immunohistochemical
expression of vascular endothelial growth
factor/vascular permeability factor in atherosclerotic intimas of human
coronary arteries. Arterioscler
Thromb Vasc Biol. 1999;19:131–139.
8. Brown MS, Goldstein JL. Lipoprotein metabolism in the macrophage: implications for cholesterol deposition in atherosclerosis. Annu Rev Biochem. 1983;52:223–261.[Medline] [Order article via Infotrieve]
9.
Steinberg D.
Oxidative modification of LDL and atherogenesis.
Circulation. 1997;95:1062–1071.
10.
Henriksen T,
Mahoney EM, Steinberg D. Enhanced macrophage degradation of low
density lipoprotein previously incubated with cultured
endothelial cells: recognition by receptors for
acetylated low density lipoproteins.
Proc Natl Acad Sci
U S A. 1981;78:6499–6503.
11.
Kita T, Nagano Y,
Yokode M, Ishii K, Kume N, Ooshima A, Yoshida H, Kawai C. Probucol
prevents the progression of atherosclerosis in Watanabe
heritable hyperlipidemic rabbit, an animal model for
familial hypercholesterolemia.
Proc Natl Acad Sci
U S A. 1987;84:5928–5931.
12. Kugiyama K, Kerns SA, Morrisett JD, Roberts R, Henry PD. Impairment of endothelium-dependent arterial relaxation by lysolecithin in modified low-density lipoproteins. Nature. 1990;344:160–162.[Medline] [Order article via Infotrieve]
13. Rajavashisth TB, Andalibi A, Territo MC, Berliner JA, Navab M, Fogelman AM, Lusis AJ. Induction of endothelial cell expression of granulocyte and macrophage colony-stimulating factors by modified low-density lipoproteins. Nature. 1990;344:254–257.[Medline] [Order article via Infotrieve]
14.
Sugiyama S,
Kugiyama K, Matsumura T, Suga S, Itoh H, Nakao K, Yasue H. Lipoproteins
regulate C-type natriuretic peptide secretion from cultured
vascular endothelial cells.
Arterioscler Thromb Vasc Biol. 1995;15:1968–1974.
15. Mangelsdorf DJ, Thummel C, Beato M, Herrlich P, Schutz G, Umesono K, Blumberg B, Kastner P, Mark M, Chambon P, et al. The nuclear receptor superfamily: the second decade. Cell. 1995;83:835–839.[Medline] [Order article via Infotrieve]
16. Tontonoz P, Hu E, Spiegelman BM. Stimulation of adipogenesis in fibroblasts by PPAR gamma 2, a lipid-activated transcription. factor. Cell. 1994;79:1147–1156.[Medline] [Order article via Infotrieve]
17. Forman BM, Tontonoz P, Chen J, Brun RP, Spiegelman BM, Evans RM. 15-Deoxy-delta 12,14-prostaglandin J2 is a ligand for the adipocyte determination factor PPAR gamma. Cell. 1995;83:803–812.[Medline] [Order article via Infotrieve]
18. Spiegelman BM, Flier JS. Adipogenesis and obesity: rounding out the big picture. Cell. 1996;87:377–389.[Medline] [Order article via Infotrieve]
19. Fujiwara T, Yoshioka S, Yoshioka T, Ushiyama I, Horikoshi H. Characterization of new oral antidiabetic agent CS-045: studies in KK and ob/ob mice and Zucker fatty rats. Diabetes. 1988;37:1549–1558.[Abstract]
20.
Nolan JJ, Ludvik
B, Beerdsen P, Joyce M, Olefsky J. Improvement in glucose tolerance and
insulin resistance in obese subjects treated with troglitazone.
N Engl J Med. 1994;331:1188–1193.
21. Saltiel AR, Olefsky JM. Thiazolidinediones in the treatment of insulin resistance and type II diabetes. Diabetes. 1996;45:1661–1669.[Abstract]
22. Tontonoz P, Nagy L, Alvarez JG, Thomazy VA, Evans RM. PPARgamma promotes monocyte/macrophage differentiation and uptake of oxidized LDL. Cell. 1998;93:241–252.[Medline] [Order article via Infotrieve]
23. Nagy L, Tontonoz P, Alvarez JG, Chen H, Evans RM. Oxidized LDL regulates macrophage gene expression through ligand activation of PPARgamma. Cell. 1998;93:229–240.[Medline] [Order article via Infotrieve]
24.
Marx N, Bourcier
T, Sukhova GK, Libby P, Plutzky J. PPARgamma activation in human
endothelial cells increases plasminogen
activator inhibitor type-1 expression:
PPARgamma as a potential mediator in vascular disease.
Arterioscler Thromb Vasc Biol. 1999;19:546–551.
25. Ohta T, Takata K, Horiuchi S, Morino Y, Matsuda I. Protective effect of lipoproteins containing apoprotein A-I on Cu2+-catalyzed oxidation of human low density lipoprotein. FEBS Lett. 1989;257:435–438.[Medline] [Order article via Infotrieve]
26. Umesono K, Murakami KK, Thompson CC, Evans RM. Direct repeats as selective response elements for the thyroid hormone, retinoic acid, and vitamin D3 receptors. Cell. 1991;65:1255–1266.[Medline] [Order article via Infotrieve]
27.
Kliewer SA, Forman
BM, Blumberg B, Ong ES, Borgmeyer U, Mangelsdorf DJ, Umesono K,
Evans RM. Differential expression and activation of a family of murine
peroxisome proliferator-activated receptors.
Proc Natl Acad Sci
U S A. 1994;91:7355–7359.
28. Niwa H, Yamamura K, Miyazaki J. Efficient selection for high-expression transfections with a novel eukaryotic vector. Gene. 1991;108:193–200.[Medline] [Order article via Infotrieve]
29.
Ribon V, Johnson
JH, Camp HS, Saltiel AR. Thiazolidinediones and insulin resistance:
peroxisome proliferator-activated receptor gamma activation
stimulates expression of the CAP gene.
Proc Natl Acad Sci
U S A. 1998;95:14751–14756.
30.
Reginato MJ,
Krakow SL, Bailey ST, Lazar MA. Prostaglandins promote and
block adipogenesis through opposing effects on peroxisome
proliferator-activated receptor gamma.
J Biol Chem. 1998;273:1855–1858.
31. Law RE, Meehan WP, Xi XP, Graf K, Wuthrich DA, Coats W, Faxon D, Hsueh WA. Troglitazone inhibits vascular smooth muscle cell growth and intimal hyperplasia. J Clin Invest. 1996;98:1897–1905.[Medline] [Order article via Infotrieve]
32.
Marx N, Schonbeck
U, Lazar MA, Libby P, Plutzky J. Peroxisome
proliferator-activated receptor gamma activators
inhibit gene expression and migration in human vascular smooth muscle
cells. Circ Res. 1998;83:1097–1103.
33. Tanaka T, Itoh H, Doi K, Fukunaga Y, Arai H, Hosoda K. Activation of PPARgamma inhibits macrophage proliferation and migration: possible therapeutic effectiveness of thiazolidinediones on diabetic vascular complications. Diabetes. 1999;48:A30. Abstract.
34. Itoh H, Doi K, Tanaka T, Fukunaga Y, Hosoda K, Inoue G, Nishimura H, Yoshimasa Y, Yamori Y, Nakao K. Hypertension and insulin resistance: role of peroxisome proliferator-activated receptor gamma. Clin Exp Pharmacol Physiol. 1999;26:558–560.[Medline] [Order article via Infotrieve]
35. Bannister AJ, Kouzarides T. The CBP co-activator is a histone acetyltransferase. Nature. 1996;384:641–643.[Medline] [Order article via Infotrieve]
36.
Zhu Y, Qi C, Jain
S, Rao MS, Reddy JK. Isolation and characterization of PBP, a protein
that interacts with peroxisome proliferator-activated receptor.
J Biol Chem. 1997;272:25500–25506.
37. Chen N-G, Sarabia SF, Malloy PJ, Zhao X-Y, Feldman D, Reaven GM. PPARg agonists enhance human vascular endothelial adhesiveness by increasing ICAM-1 expression. Biochem Biophys Res Commun. 1999;263:718–722.[Medline] [Order article via Infotrieve]
38. Inoue M, Itoh H, Ueda M, Naruko T, Kojima A, Chun T-H, Ogawa Y, Igaki T, Yamashita J, Doi K, et al. Expression of vascular endothelial growth factor (VEGF) in human coronary atherosclerosis and its gene regulation by oxidized LDL (Ox-LDL): VEGF as a possible progression factor for human atherosclerosis. Circulation. 1997;96(suppl I):I-2. Abstract.
39. Yonemitsu Y, Kaneda Y, Morishita R, Kazunori N, Nakashima Y, Sueishi K. Characterization of in vivo gene transfer into the arterial wall mediated by the Sendai virus (hemagglutinating virus of Japan) liposomes: an effective tool for the in vivo study of arterial diseases. Lab Invest. 1996;75:313–323.[Medline] [Order article via Infotrieve]
40. Sueishi K, Yonematsu Y, Nakagawa K, Kaneda Y. Atherosclerosis and angiogenesis: its pathophysiological significance in humans as well as in an animal model induced by gene transfer of vascular endothelial growth factor (VEGF). In: Abstracts of the Fourth Saratoga International Conference on Atherosclerosis, Hawaii, February 6–8, 1996. Abstract SV-4.
41.
De Boer OJ, Van
der Wal AC, Teeling P, Becker AE. Leucocyte recruitment in rupture
prone regions of lipid-rich plaques: a prominent role for
neovascularization? Cardiovasc
Res. 1999;41:443–449.
42. Spyridopoulos I, Brogi E, Kearney M, Sullivan AB, Cetrulo C, Isner JM, Losordo DW. Vascular endothelial growth factor inhibits endothelial cell apoptosis induced by tumor necrosis factor-alpha: balance between growth and death signals [published erratum appears in J Mol Cell Cardiol. 1998;30:897]. J Mol Cell Cardiol. 1997;29:1321–1330.
This article has been cited by other articles:
 |
|
 |
|
  J. Barlic, W. Zhu, and P. M. Murphy Atherogenic Lipids Induce High-Density Lipoprotein Uptake and Cholesterol Efflux in Human Macrophages by Up-Regulating Transmembrane Chemokine CXCL16 without Engaging CXCL16-Dependent Cell Adhesion J. Immunol., June 15, 2009; 182(12): 7928 - 7936. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 V. Mallika, B. Goswami, and M. Rajappa Atherosclerosis Pathophysiology and the Role of Novel Risk Factors: A Clinicobiochemical Perspective Angiology, November 1, 2007; 58(5): 513 - 522. [Abstract] [PDF]
|
|
|
|
 |
|
 J. L. Mehta Oxidized or Native Low-Density Lipoprotein Cholesterol: Which Is More Important in Atherogenesis? J. Am. Coll. Cardiol., September 5, 2006; 48(5): 980 - 982. [Full Text] [PDF]
|
|
|
|
 |
|
 A. Alfranca, M. A. Iniguez, M. Fresno, and J. M. Redondo Prostanoid signal transduction and gene expression in the endothelium: Role in cardiovascular diseases Cardiovasc Res, June 1, 2006; 70(3): 446 - 456. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Dulak, S. P Schwarzacher, R. H Zwick, H. Alber, G. Millonig, C. Weiss, H. Hugel, M. Frick, A. Jozkowicz, O. Pachinger, et al. Effects of local gene transfer of VEGF on neointima formation after balloon injury in hypercholesterolemic rabbits Vascular Medicine, November 1, 2005; 10(4): 285 - 291. [Abstract] [PDF]
|
|
|
|
 |
|
 W. Song, J. L. Barth, Y. Yu, K. Lu, A. Dashti, Y. Huang, C. K. Gittinger, W. S. Argraves, and T. J. Lyons Effects of Oxidized and Glycated LDL on Gene Expression in Human Retinal Capillary Pericytes Invest. Ophthalmol. Vis. Sci., August 1, 2005; 46(8): 2974 - 2982. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. Hamerman Osteoporosis and atherosclerosis: biological linkages and the emergence of dual-purpose therapies QJM, July 1, 2005; 98(7): 467 - 484. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. Patel, K. L Wyne, and D. K McGuire Thiazolidinediones, peripheral oedema and congestive heart failure: what is the evidence? Diabetes and Vascular Disease Research, May 1, 2005; 2(2): 61 - 66. [Abstract] [PDF]
|
|
|
|
|
|
J. A. Rodriguez, B. Nespereira, M. Perez-Ilzarbe, E. Eguinoa, and J. A. Paramo Vitamins C and E prevent endothelial VEGF and VEGFR-2 overexpression induced by porcine hypercholesterolemic LDL Cardiovasc Res, February 15, 2005; 65(3): 665 - 673. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. Friday, J. Glass, and F. Turturro Troglitazone Exerts a Cell-Dependent Effect on the Extracellular Secretion of VEGF165 Independently from the Rapamycin Sensitivity or the Level of eIF4E Expression in AML-Derived Cells. Blood (ASH Annual Meeting Abstracts), November 16, 2004; 104(11): 4363 - 4363. [Abstract]
|
|
|
|
 |
|
 M. Negishi, H. Shimizu, S. Okada, A. Kuwabara, F. Okajima, and M. Mori 9HODE Stimulates Cell Proliferation and Extracellular Matrix Synthesis in Human Mesangial Cells via PPAR{gamma} Experimental Biology and Medicine, November 1, 2004; 229(10): 1053 - 1060. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. Grau, M. A. Iniguez, and M. Fresno Inhibition of Activator Protein 1 Activation, Vascular Endothelial Growth Factor, and Cyclooxygenase-2 Expression by 15-Deoxy-{Delta}12,14-Prostaglandin J2 in Colon Carcinoma Cells: Evidence for a Redox-Sensitive Peroxisome Proliferator-Activated Receptor-{gamma}-Independent Mechanism Cancer Res., August 1, 2004; 64(15): 5162 - 5171. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. R. Chade, M. D. Bentley, X. Zhu, M. Rodriguez-Porcel, S. Niemeyer, B. Amores-Arriaga, C. Napoli, E. L. Ritman, A. Lerman, and L. O. Lerman Antioxidant Intervention Prevents Renal Neovascularization in Hypercholesterolemic Pigs J. Am. Soc. Nephrol., July 1, 2004; 15(7): 1816 - 1825. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. M Ridker, N. J. Brown, D. E. Vaughan, D. G. Harrison, and J. L. Mehta Established and Emerging Plasma Biomarkers in the Prediction of First Atherothrombotic Events Circulation, June 29, 2004; 109(25_suppl_1): IV-6 - IV-19. [Full Text] [PDF]
|
|
|
|
 |
|
 R. Walczak, S. B. Joseph, B. A. Laffitte, A. Castrillo, L. Pei, and P. Tontonoz Transcription of the Vascular Endothelial Growth Factor Gene in Macrophages Is Regulated by Liver X Receptors J. Biol. Chem., March 12, 2004; 279(11): 9905 - 9911. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 I. E. Konstantinov, J. G. Coles, C. Boscarino, M. Takahashi, J. Goncalves, J. Ritter, and G. S. Van Arsdell Gene expression profiles in children undergoing cardiac surgery for right heart obstructive lesions J. Thorac. Cardiovasc. Surg., March 1, 2004; 127(3): 746 - 754. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D.-H. Cho, Y. J. Choi, S. A. Jo, and I. Jo Nitric Oxide Production and Regulation of Endothelial Nitric-oxide Synthase Phosphorylation by Prolonged Treatment with Troglitazone: EVIDENCE FOR INVOLVEMENT OF PEROXISOME PROLIFERATOR-ACTIVATED RECEPTOR (PPAR) {gamma}-DEPENDENT AND PPAR{gamma}-INDEPENDENT SIGNALING PATHWAYS J. Biol. Chem., January 23, 2004; 279(4): 2499 - 2506. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. F. Klein, J. Allard, Z. Avnur, T. Nikolcheva, D. Rotstein, A. S. Carlos, M. Shea, R. V. Waters, J. K. Belknap, G. Peltz, et al. Regulation of Bone Mass in Mice by the Lipoxygenase Gene Alox15 Science, January 9, 2004; 303(5655): 229 - 232. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. K. Lutgendorf, S. Cole, E. Costanzo, S. Bradley, J. Coffin, S. Jabbari, K. Rainwater, J. M. Ritchie, M. Yang, and A. K. Sood Stress-Related Mediators Stimulate Vascular Endothelial Growth Factor Secretion by Two Ovarian Cancer Cell Lines Clin. Cancer Res., October 1, 2003; 9(12): 4514 - 4521. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Fu, J. Zhang, Y. Lin, X. Zhu, M. U. Ehrengruber, and Y. E. Chen Early Growth Response Factor-1 Is a Critical Transcriptional Mediator of Peroxisome Proliferator-activated Receptor-gamma 1 Gene Expression in Human Aortic Smooth Muscle Cells J. Biol. Chem., July 19, 2002; 277(30): 26808 - 26814. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Fauconnet, I. Lascombe, E. Chabannes, G.-L. Adessi, B. Desvergne, W. Wahli, and H. Bittard Differential Regulation of Vascular Endothelial Growth Factor Expression by Peroxisome Proliferator-activated Receptors in Bladder Cancer Cells J. Biol. Chem., June 21, 2002; 277(26): 23534 - 23543. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Franz Alber, J. Dulak, M. Frick, W. Dichtl, S. Paul Schwarzacher, O. Pachinger, and F. Weidinger Atorvastatin decreases vascular endothelial growth factor in patients with coronary artery disease J. Am. Coll. Cardiol., June 19, 2002; 39(12): 1951 - 1955. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. Molavi, N. Rasouli, and J. L. Mehta Peroxisome Proliferator-Activated Receptor Ligands as Antiatherogenic Agents: Panacea or Another Pandora's Box? Journal of Cardiovascular Pharmacology and Therapeutics, March 1, 2002; 7(1): 1 - 8. [Abstract] [PDF]
|
|
|
|
|
|
J. Dulak, A. Jozkowicz, M. Frick, H. F. Alber, W. Dichtl, S. P. Schwarzacher, O. Pachinger, and F. Weidinger Vascular endothelial growth factor: angiogenesis, atherogenesis or both? J. Am. Coll. Cardiol., December 1, 2001; 38(7): 2137 - 2138. [Full Text] [PDF]
|
|
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||