(Circulation. 2000;102:806.)
© 2000 American Heart Association, Inc.
Basic Science Reports |
Increased Activity of Nuclear Factor-
B Participates in Cardiovascular Remodeling Induced by Chronic Inhibition of Nitric Oxide Synthesis in Rats
From the Departments of Cardiovascular Medicine (S.K., K.E., C.K., M. Koyanagi, H.S., A.T.) and Pathology (K.S.), Graduate School of Medical Science, Kyushu University, Fukuoka; Division of Gene Therapy Sciences, Osaka University Medical School, Osaka (R.M., Y.K.); and Discovery Research Laboratory, Tanabe Sei-Yaku Co Ltd, Saitama (M. Katoh), Japan.
Correspondence to Kensuke Egashira, MD, PhD, Department of Cardiovascular Medicine, Graduate School of Medical Sciences, Kyushu University, 3-1-1, Maidashi, Higashi-ku, Fukuoka 812-8582, Japan. E-mail egashira{at}cardiol.med.kyushu-u.ac.jp
 |
Abstract |
|---|
 Top Abstract IntroductionMethodsResultsDiscussionReferences |
|---|
Background—Chronic inhibition of endothelial nitric oxide (NO) synthesis by the administration of N
-nitro-L-arginine methyl
ester (L-NAME) to rats induces early vascular inflammatory changes
[monocyte infiltration into coronary vessels, nuclear
factor-
B (NF-B) activation, and monocyte chemoattractant
protein-1 expression] as well as subsequent
arteriosclerosis (medial thickening and
perivascular fibrosis) and cardiac fibrosis. However, no direct
evidence for the importance of NF-B in this process is
known.
Methods and Results—We examined the effect of a
cis element decoy strategy to address the functional
importance of NF-B in the pathogenesis of
cardiovascular remodeling. We found here that in vivo
transfection of cis element decoy
oligodeoxynucleotides against NF-B to hearts prevented
the L-NAME–induced early inflammation and subsequent coronary
vascular medial thickening. In contrast, NF-B decoy
oligodeoxynucleotide transfection did not decrease the
development of fibrosis, the expression of transforming growth
factor-ß1 mRNA, or systolic pressure overload
induced by L-NAME administration.
Conclusions—The NF-B system participates importantly in the
development of early vascular inflammation and subsequent medial
thickening but not in fibrogenesis in this model. The present study
may provide a new aspect of how endothelium-derived NO
contributes to anti-inflammatory and/or
antiarteriosclerotic properties of the vascular
endothelium in vivo.
Key Words: endothelium-derived factors • inflammation • proteins • cells • nitric oxide • nuclear factor-B
|
Introduction |
|---|
|
TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
The vascular endothelium becomes dysfunctional in the early stages of vascular diseases.1 2 3 Such endothelial abnormalities are associated with reduced activity of nitric oxide (NO) and increased expression of inflammation-promoting genes.4 5 6 7 8 Recently, endothelium-derived NO has been recognized to be an anti-inflammatory and antiarteriosclerotic molecule. Mice lacking endothelial-type NO synthase exhibit hypertension and an enhanced vascular remodeling in response to injury.9 10 We recently reported that chronic inhibition of NO synthesis by the administration of N
-nitro-L-arginine
methyl ester (L-NAME) induces early inflammation [monocyte
infiltration, monocyte chemoattractant protein-1 (MCP-1) expression,
and nuclear factor-B (NF-B) activation] and late
cardiovascular remodeling in rats (Figure 1
).11 12 13 14 15 We also
demonstrated that normalization of arterial hypertension by
hydralazine treatment did not attenuate such pathological
changes,11 13 16 17 suggesting a minor role of systemic
arterial hypertension in the development of such
pathological changes in this model.
|
NF-B is presumed to be an important redox-sensitive transcriptional
factor that regulates transcription of genes encoding inflammatory
cytokines, adhesion molecules, and chemokines.18
NF-B is activated by a battery of stimuli leading to
atherogenic events, including oxidative stress, tumor necrosis factor,
and angiotensin II.19 20 21 MCP-1 is a member of
the C-C chemokines and a potent chemotactic factor for
monocytes.22 Increased activity of NF-B23
or MCP-124 has been observed in
arteriosclerotic and/or atherosclerotic lesions. We
recently demonstrated that increased superoxide anion production
contributes to the pathogenesis of ACE activation and that ACE
inhibition, angiotensin II type 1 receptor blockade, or
antioxidant therapy with N-acetylcysteine prevents vascular
inflammation, MCP-1 expression, NF-B activation, and subsequent
remodeling in the rat model (Figure 1
).25
These observations suggest that the increase in local
angiotensin II activity plays a primary role in the
development of such early and late cardiovascular
pathological changes (Figure 1). It is also suggested that
increased activity of NF-B would participate in the inflammatory
changes through transcription of MCP-1 in the rat model of chronic
inhibition of NO synthesis (Figure 1). However, no direct
evidence for the functional importance of NF-B in the formation of
such cardiovascular inflammation and remodeling has
been addressed.
To achieve effective blockade of NF-B activity in vivo, transfection
of a cis element decoy against the NF-B binding site
seems to be a useful strategy.26 Recently, Morishita
et al27 elegantly innovated the cis element
decoy strategy as an effective and feasible in vivo gene therapy. They
reported that transfection of NF-B decoy
oligodeoxynucleotides (ODNs) to rat hearts by injection of
hemagglutinating virus of Japan (HVJ)–liposome complex in the
ascending aorta markedly reduced myocardial injury due to myocardial
infarction. Thus, we used this novel strategy and investigated the role
of NF-B in the development of early inflammatory changes and late
cardiovascular remodeling in a rat model of chronic
inhibition of NO synthesis.
|
Methods |
|---|
|
TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
Decoy ODN Sequences and Preparation of HVJ-Liposome Complexes
The sequences of NF-
B decoy ODNs and scramble decoy ODNs used
in this study were as follows (consensus sequences of binding site for
NF-B are italicized): NF-B decoy ODN:
5'-CCTTGAAGGGATTTCCCTCC-3';
3'-GGAACTTCCCTAAAGG-GAGG-5'. Scramble
decoy ODN: 5'-TTGCCGTACCTGACT-TAGCC-3';
3'-AACGGCATGGACTGAATCGG-5'.
The NF-B decoy ODNs, but not scramble decoy ODNs, have been shown to
bind the NF-B transcriptional factor.27 HVJ-liposome
complexes were prepared as described.27 The final
concentration of decoy ODNs was 15 µmol/L.
In Vivo Transfection of ODNs Into Hearts
After a rat was anesthetized with
intraperitoneal pentobarbital, the right common
carotid artery was surgically exposed. A balloon catheter 1.5 mm
in diameter was introduced into the common carotid artery and advanced
to the ascending aorta. After the catheter tip was positioned at the
aortic sinus of Valsalva, the balloon was inflated, and HVJ-liposome
complex containing FITC-labeled or unlabeled ODNs (1 mL at 4°C) was
infused selectively into the coronary arteries. Then the
catheter was removed, the wound was closed, and the animals were
allowed to recover from the surgery.
Animal Model of Chronic Inhibition of NO Synthesis
The present study protocol was reviewed and approved by the
Committee on Ethics on Animal Experiments, Kyushu University Faculty of
Medicine, and the experiments were conducted according to the
Guidelines for Animal Experiments of Kyushu University Faculty of
Medicine.
Four groups of male Wistar-Kyoto rats were studied. The control group
received normal water and chow. The L-NAME group received L-NAME in
drinking water (1 mg/mL). The L+NF group received L-NAME 2 days after
NF-B decoy transfection. The L+SD group received L-NAME 2 days after
scramble decoy transfection.
On day 3 after L-NAME administration was begun, systolic blood pressures (by the tail-cuff method) were measured. Then the rats were euthanized for morphometric, immunohistochemical, and biochemical analyses. Furthermore, some rats in each group received L-NAME for 7 days and untreated water during the following 21 days. On day 28, they were euthanized for morphometric analysis.
Electrophoretic Mobility Shift Assays
To determine the increase in NF-B binding to the nucleus,
electrophoretic mobility shift assays were performed. Five rats in each
group were used. Nuclear extracts were prepared from the whole-heart
homogenates as described.15 The NF-B
oligonucleotides corresponding to putative consensus
sequences (NF-B: 5'-AGTTGAGGGGACTTTCCCAGGC-3') (Promega
Biotechnology Inc) were labeled with
[
-32P]ATP and T4 polynucleotide
kinase and purified by Sephadex G-50 column (Pharmacia Biotechnology
Inc). Nuclear extract (10 µg) was incubated with
1x105 cpm of labeled probe and 2 µg of
poly(dI-dC) in a buffer containing 10 mmol/L Tris-HCl (pH 7.5),
1 mmol/L EDTA, 4% glycerol, 100 mmol/L NaCl, 5 mmol/L
DTT, and 100 g/L BSA for 30 minutes at room temperature (25°C). Then
the samples were electrophoresed on 5% acrylamide/0.5xTBE
gel (1xTBE: 90 mmol/L Tris borate, 2 mmol/L EDTA). After
electrophoresis, gels were dried and subjected to
autoradiography. Autoradiographs were later subjected
to laser densitometry. Specificity was determined by the addition of
excess cold oligonucleotide to the nuclear extracts 10
minutes before addition of radiolabeled probe.
Northern Blot Analysis
Total RNA was extracted from each sample, poly(A)+RNA was
purified, and then Northern blot hybridization was performed as we
described.15 The cDNA probes used were as follows: a rat
MCP-1,28 a rat transforming growth factor
(TGF)-ß1 cDNA (a gift from Dr T. Nakamura,
Department of Biochemistry, Kyushu University), and a mouse GAPDH
(American Type Culture Collection). Relative amounts of MCP-1 and
TGF-ß1 mRNA were normalized against the amounts
of GAPDH mRNA.
Histopathology and Immunohistochemistry
Tissue sections were prepared as described.14 15
After fixation with methacarn solution, the left ventricle was
separated from the atria, right ventricle, and great vessels. The left
ventricle was cut into 5 pieces perpendicular to the long axis. All
tissues were dehydrated, embedded in paraffin, cut into slices 5
µm thick, and mounted on slides. The sections were stained with
either hematoxylin-eosin, Masson’s trichrome, or
immunostaining with antibodies against
macrophage/monocyte (ED1, Serotec), proliferating cell nuclear
antigen (PCNA) (Dako), nonimmune mouse IgG (Zymed), or p50/NF-B
(Santa Cruz). The slides were washed and incubated with biotinylated,
affinity-purified goat anti-mouse IgG. After avidin-biotin
amplification, the slides were incubated with 3',3'-diaminobenzidine
and counterstained with hematoxylin.
Morphometry and cell counting were performed by a single observer who was blind to the treatment protocols as described.15 On day 3, to quantify the number of immunopositive cells in hearts, each section (5 per heart) immunohistochemically stained by an antibody against ED1 or PCNA was scanned at x100 magnification. The number of positive cells in each section was counted, and the average number of positive cells per section was reported in each heart.
On day 28, to evaluate the thickening of the coronary arterial wall and the extent of perivascular fibrosis, short-axis images of the coronary artery were analyzed. The inner border of the lumen and the outer border of the tunica media were traced from Masson’s trichrome–stained sections at x100 to x200 magnification. The wall-to-lumen ratio (the ratio of medial thickness to the internal diameter) and the area of fibrosis (area of collagen deposition stained with aniline blue) immediately surrounding the blood vessel were then calculated. Perivascular fibrosis was estimated as the ratio of the area of fibrosis surrounding the vessel wall to the total vessel area. Myocardial reparative fibrosis after myocyte necrosis was also determined. Areas of myocardial necrosis replaced by fibrosis were calculated as the total area of fibrosis in the entire visual field divided by the total area of connective tissue and myocardium in the visual field.
Analysis of Fluorescence Isothiocyanate
ODNs
The efficacy of FITC-labeled ODNs delivered by the HVJ-liposome
method was assessed with a fluorescence microscope. Serial
tissue sections of hearts injected with HVJ-liposome complex with
FITC-labeled or unlabeled ODNs were prepared. One section was stained
with propidium iodide to identify the nuclei. We used a 2-color
fluorescence approach, which allows the identification of cells
with nuclear incorporation of transfected ODNs. In addition, another
section was stained with hematoxylin-eosin to differentiate cell types
stained with FITC and/or propidium iodide.
Measurements of ACE Activity
Cardiac tissues were isolated, and the ACE activity was measured
by fluorometric assay as described.11 12 Tissue ACE
activity was calculated as nanomoles His-Leu generated per
milligram tissue weight per hour.
Statistical Analysis
Data are expressed as mean±SEM. Statistical analysis of
differences was compared by ANOVA and Bonferroni’s multiple comparison
test. A level of P<0.05 was considered statistically
significant.
|
Results |
|---|
|
TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
Systolic Arterial Pressure
Compared with the control group, the L-NAME groups showed a rise in systolic arterial pressure on days 3 and 28 of treatment. This increase in systolic blood pressure was not influenced by NF-
B decoy transfection or scramble decoy transfection
(Table).
|
In Vivo Transfection of Decoy ODNs Into Heart
Electrophoretic mobility shift assay showed that the NF-B
binding affinity was markedly increased in the hearts from the L-NAME
group on day 3 of treatment (Figure 2A).
The increased NF-B activity was suppressed by NF-B decoy ODN
transfection but not by scramble decoy ODN transfection in vivo (Figure
2A). One day after intracoronary infusion of
FITC-labeled ODNs, we examined the location of delivered decoy ODNs
with the fluorescence microscope. The fluorescence was
localized intensely in the nuclei (yellow) as well as cytoplasm (green)
of vascular endothelial cells. Weak to moderate
fluorescence (orange) was noted in the nuclei in vascular
smooth muscle cells, cardiomyocytes, and some
interstitial cells (Figure 2B).
|
Effects of the Transfection of NF-B Decoy ODNs on the
Inflammatory and Proliferative Changes and the mRNA Levels of MCP-1 and
TGF-ß1 on Day 3
We observed no evidence of inflammation in the control group
(Figure 3A). Attachment of mononuclear
leukocytes to the endothelium of coronary
vessels was seen in the L-NAME (data not shown) and L+SD groups (Figure 3A). A marked infiltration of mononuclear leukocytes in the
perivascular areas immediately surrounding the coronary
arteries and veins and the myocardial interstitial spaces
was observed in those 2 groups (Figure 3A). The majority of
leukocytes that had infiltrated into the lesions were found to be
ED1-positive monocytes (Figure 3A). Nuclear staining for PCNA
antibody was observed in some endothelial cells,
vascular smooth muscle cells in the media, monocytes, or
myofibroblast-like cells (Figure 3A). We also examined
localization of NF-B activation by immunohistochemistry (Figure 3B). Compared with the control group, the translocation of
p50/NF-B immunoreactivity to the nucleus, from faint cytoplasmic
staining to a prominent nuclear pattern, was observed in the
endothelial cells, some smooth muscle cells in the
media, and infiltrated inflammatory cells (monocytes and
myofibroblasts) in the L-NAME group. This change suggests that
redistribution of p50/NF-B from the cytoplasm to the nucleus has
occurred. Such translocation of NF-B immunoreactivity was observed
in some areas in the myocardial myocyte to a similar extent in the
control and L-NAME groups. These results suggest that NF-B
activation may occur predominantly in coronary vessels.
|
When ED1-positive monocytes or PCNA-positive cells were counted by use
of immunohistochemistry, the number of immunopositive cells per section
was significantly greater in the L-NAME group than in the control group
(Figure 4A and 4B). These inflammatory
and proliferative changes were markedly reduced by NF-B decoy ODN
transfection but not by scramble decoy ODN transfection (Figure 4A and 4B).
|
P<0.01 vs
L-NAME group, #P<0.01 vs L+SD group. C, Northern blot
analysis, representative
autoradiogram of expression of cardiac MCP-1,
TGF-ß1, and GAPDH mRNA in rats from control group, L-NAME
group, L+NF group, and L+SD group. D, Densitometric analysis of
data in C (n=6). Expression of MCP-1 and TGF-ß1 mRNA in
each sample is normalized relative to GAPDH mRNA expression in that
sample. *P<0.01 vs control group,
We examined the expression of MCP-1 and TGF-ß1
mRNA in the heart (Figure 4C and 4D). The cardiac MCP-1 and
TGF-ß1 mRNA levels were significantly increased
in the L-NAME group. The increased expression of MCP-1 mRNA was
significantly reduced in the L+NF group but not in the L+SD group. In
contrast, the increased expression of TGF-ß1
mRNA was not reduced by NF-B decoy or scramble decoy ODN
transfection (Figure 4C and 4D).
Effects of the Transfection of NF-B Decoy on
Cardiovascular Remodeling on Day 28
The increase in the medial thickening (the wall-to-lumen ratio) of
coronary arteries seen in the L-NAME group was prevented in the
L+NF group but not in the L+SD group (Figure 5). In contrast, the increases in
perivascular and cardiac fibrosis were not affected by NF-B decoy or
scramble decoy transfection (Figure 5). We did not examine left
ventricular hypertrophy or function, because no
such changes are evident within 28 days of
treatment.11
|
Tissue ACE Activity
Cardiac tissue ACE activity was increased in the L-NAME group
(Table). This increased ACE activity was not influenced by
NF-B decoy ODN transfection or scramble decoy ODN transfection
(Table).
|
Discussion |
|---|
|
TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
The novel findings that have emerged from the present study are that in vivo transfection of cis element decoy against NF-
B binding site to the heart markedly suppressed NF-B activity
and prevented early inflammatory changes. Furthermore, transfection of
NF-B decoy ODNs inhibited the late development of coronary
vascular medial thickening after inhibition of NO synthesis. The
observed effect of NF-B decoy transfection was independent of
arterial hypertension induced by L-NAME administration. Our
present findings suggest that the NF-B system may be essential
in the development of early inflammation and subsequent
coronary vascular medial thickening in our model.
In the present study, we could transfect the decoy ODNs
superselectively into coronary arteries and demonstrate that
the strategy achieved NF-B blockade in rat hearts (Figure 2).
Thus, our present observations suggest that increased activity of
NF-B participates essentially in the L-NAME–induced increases in
MCP-1 expression and subsequent monocyte recruitment. This claim is
supported by in vitro evidence that cis-acting elements for
the NF-B binding site in the MCP-1 gene promoter region are
responsible for increased transcription of the MCP-1
gene29 and that inhibition of NO synthesis increases
the activity of NF-B in vitro.5 6 7 8 We previously
demonstrated the increased production of MCP-1 protein in
endothelial cells, medial smooth muscle cells, and
infiltrating inflammatory cells.14 15 Because the NF-B
activation in immunohistochemistry was also observed in these cells
(Figure 3B), it is likely that increased NF-B–mediated
transcription and protein production of MCP-1 participate
essentially in the development of inflammatory changes in our
model.
Transfection of NF-B decoy ODNs also inhibited the increase in PCNA
(a marker of cell proliferation)-positive cells. Nuclear staining for
PCNA antibody was observed in endothelial cells,
vascular smooth muscle cells in the media, infiltrating monocytes, and
myofibroblasts.14 15 These results suggest that increased
NF-B–mediated transcription of inflammation-promoting genes,
including MCP-1, activated those cells and thus induced the
vascular proliferative changes. These proliferating cells can secrete
growth-promoting factors such as platelet-derived growth factor,
fibroblast growth factor, and reactive oxygen species.1 3
Therefore, we hypothesize that NF-B–mediated transcription of MCP-1
induced the recruitment of monocytes and activated vascular
smooth muscle cells and monocytes during the early phase, which in turn
caused proliferation of vascular smooth muscles by producing those
growth-promoting factors. Transfection of NF-B decoy ODNs thereby
inhibited the development of vascular medial thickening during the late
phase in the present study. A recent study demonstrated that MCP-1
may directly stimulate proliferation and migration of cultured vascular
smooth muscle cells.30 Hence, it is reasonable to assume
that the NF-B system plays a key role in early vascular inflammation
and subsequent vascular medial thickening after blockade of NO
synthesis in vivo.
Despite a marked inhibition of monocyte infiltration, transfection of
NF-B decoy ODNs could not reduce gene expression of
TGF-ß1 as well as perivascular and cardiac
fibrosis. TGF-ß1 may induce transformation of
fibroblasts to myofibroblasts, stimulate production of
extracellular matrix proteins, and thus play an central role in tissue
fibrosis.31 There are no NF-B binding sites in the
promoter region of the TGF-ß1 gene. Thus, the
NF-B system may not be involved in cardiac fibrogenesis in our
model.
We previously demonstrated that local activity of ACE plays an
important role in early cardiovascular inflammation and
late remodeling in our model.9 13 In this study, there was
no significant difference in the enzyme activity between hearts from
the L-NAME, L+NF, and L+SD group. Thus, it is likely that the observed
effects of NF-B transfection were independent of local ACE
activity.
We previously demonstrated that pharmacological inhibition of endothelial NO synthesis by L-NAME administration caused early cardiovascular inflammation and subsequent remodeling in rats. This conclusion is supported by our previous reports demonstrating that (1) endothelial NO production was blunted in the aorta from rats that received L-NAME but not from those that received D-NAME,14 (2) treatment with L-arginine prevented the L-NAME–induced inflammation and remodeling,13 14 17 and (3) administration of D-NAME did not induce such cardiovascular pathological changes.17 However, such pathological changes as seen in the rat model have not been described in genetically mutant mice lacking the endothelial NO synthase gene.9 10 These mice exhibit greater inflammatory and proliferative vascular responses to injury. The mechanisms of the difference between the rat and mouse models is probably multifactorial. One plausible explanation is that a defective endothelial NO synthase gene since birth might be compensated by other genes, so that no inflammatory or proliferative vascular changes have been observed at rest in mice lacking the endothelial NO synthase gene. In contrast, postnatal blockade of NO synthesis by administration of L-NAME to adult rats caused early inflammatory vascular responses and subsequent remodeling, as seen in the present study. Thus, these observations in the L-NAME–treated rats as well as in the mice lacking endothelial NO synthase may have clinically relevant implications, in that endothelium-derived NO is recognized to be an endogenous anti-inflammatory and/or antiarteriosclerotic factor.
In conclusion, the present study has provided direct in vivo
evidence of the role of the NF-B system in the development of
vascular medial thickening at least by inducing MCP-1 in the rat model
of chronic inhibition of NO synthesis. Although we did not examine
whether other cis elements of the promoter region, such as
AP-1 and SP-1, are involved in the regulation of MCP-1 expression in
our model, the present study may provide a new aspect of how
endothelium-derived NO contributes to anti-inflammatory
and/or antiarteriosclerotic properties of the
vascular endothelium in vivo. It appears that NO
decreases monocyte recruitment to the arterial wall at
least by suppressing NF-B–mediated transcription of MCP-1 in vivo.
Thus, preservation of normal endothelial NO activity
may be an effective therapeutic strategy to reduce
cardiovascular ischemic events in patients with
vascular diseases. It might be of clinical interest to know whether an
NF-B decoy strategy can reduce restenosis after
coronary angioplasty or the vulnerability of an atherosclerotic
plaque prone to rupture.
|
Acknowledgments |
|---|
This study was supported by Grants-in-Aid for Scientific Research (11470164, 11158216, 11557056, 10307019, 10177226) from the Ministry of Education, Science, and Culture, Tokyo; by the Ryouichi Naito Foundation for Medical Research, Osaka; and by a Research Grant from the Kanae Foundation of Research for New Medicine, Osaka, Japan.
Received February 9, 2000; revision received March 15, 2000; accepted March 16, 2000.
|
References |
|---|
|
TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
1. Ross R. Atherosclerosis: an inflammatory disease. N Engl J Med. 1999;340:115–126.
2. Harrison DG. Cellular and molecular mechanisms of endothelial cell dysfunction. J Clin Invest. 1997;100:2153–2157.[Medline] [Order article via Infotrieve]
3. Griendling KK, Alexander RW. Endothelial control of the cardiovascular system: recent advances. FASEB J. 1996;10:283–292.[Abstract]
4.
von der Leyen HE, Gibbons GH, Morishita R, et
al. Gene therapy inhibiting neointimal vascular
lesion: in vivo transfer of endothelial cell nitric
oxide synthase gene. Proc Natl Acad Sci U S A. 1995;92:1137–1141.
5.
Tsao PS, Wang B, Buitrago R, et al. Nitric oxide
regulates monocyte chemotactic protein-1. Circulation. 1997;96:934–940.
6.
Zeiher AM, Fisslthaler B, Schray-Utz B, et al. Nitric
oxide modulates the expression of monocyte chemoattractant protein 1 in
cultural human endothelial cells. Circ Res. 1995;76:980–986.
7.
Peng HB, Libby P, Liao JK. Induction and stabilization
of I kappa B alpha by nitric oxide mediates inhibition of NF-kappa B.
J Biol Chem. 1995;270:14214–14219.
8. De Caterina R, Libby P, Peng HB, et al. Nitric oxide decreases cytokine-induced endothelial activation: nitric oxide selectively reduces endothelial expression of adhesion molecules and proinflammatory cytokines. J Clin Invest. 1995;96:60–68.
9. Moroi M, Zhang L, Yasuda T, et al. Interaction of genetic deficiency of endothelial nitric oxide, gender, and pregnancy in vascular response to injury in mice. J Clin Invest. 1998;101:1225–1232.[Medline] [Order article via Infotrieve]
10. Rudic RD, Shesely EG, Maeda N, et al. Direct evidence for the importance of endothelium-derived nitric oxide in vascular remodeling. J Clin Invest. 1998;101:731–736.[Medline] [Order article via Infotrieve]
11. Takemoto M, Egashira K, Usui M, et al. Important role of tissue angiotensin-converting enzyme activity in the pathogenesis of coronary vascular and myocardial structural changes induced by long-term blockade of nitric oxide synthesis in rats. J Clin Invest. 1997;99:278–287.[Medline] [Order article via Infotrieve]
12.
Takemoto M, Egashira K, Tomita H, et al. Chronic
angiotensin-converting enzyme inhibition and
angiotensin II type 1 receptor blockade.
Hypertension. 1997;30:1621–1627.
13.
Katoh M, Egashira K, Usui M, et al. Cardiac
angiotensin II receptor is upregulated by long-term
inhibition of nitric oxide synthesis in rats. Circ Res. 1998;83:743–751.
14.
Tomita H, Egashira K, Kubo-Inoue M, et al. Inhibition
of nitric oxide synthesis induces inflammatory changes and monocyte
chemoattractant protein-1 expression in rat hearts and vessels.
Arterioscler Thromb Vasc Biol. 1998;18:1456–1464.
15.
Usui M, Egashira K, Tomita H, et al. Important role of
local angiotensin II activity mediated via type 1 receptor
in the pathogenesis of cardiovascular inflammatory
changes induced by chronic blockade of nitric oxide synthesis in rats.
Circulation. 2000;101:305–310.
16. Numaguchi K, Egashira K, Takemoto M, et al. Chronic inhibition of nitric oxide synthesis causes pathologic coronary microvascular remodeling and myocardial fibrosis. Hypertension. 1995;26(pt 1):957–962.
17.
Tomita H, Egashira K, Ohara Y, et al. Early induction
of transforming growth factor-ß via angiotensin II
type 1 receptors contributes to cardiac fibrosis induced by long-term
blockade of nitric oxide synthesis in rats. Hypertension. 1998;32:273–279.
18.
Collins T, Read MA, Neish AS, et al. Transcriptional
regulation of endothelial cell adhesion molecules:
NF-B and cytokine-inducible enhancers. FASEB
J. 1995;9:899–909.[Abstract]
19. Marui N, Offermann MK, Swerlick R, et al. Vascular cell adhesion molecule-1 (VCAM-1) gene transcription and expression are regulated through an antioxidant-sensitive mechanism in human vascular endothelial cells. J Clin Invest. 1993;92:1866–1874.
20. Satriano JA, Shuldiner M, Hora K, et al. Oxygen radical as second messenger for expression of the monocyte chemoattractant protein, JE/MCP-1, and the monocyte colony-stimulating factor, CSF-1, in response to tumor necrosis factor-alpha and immunoglobulin G. J Clin Invest. 1993;92:1564–1571.
21.
Hernandez-Presa M, Bustos C, Ortego M, et al.
Angiotensin-converting enzyme inhibition prevents
arterial nuclear factor-kappa B activation, monocyte
chemoattractant protein-1 expression, and macrophage
infiltration in a rabbit model of early accelerated
atherosclerosis. Circulation. 1997;95:1532–1541.
22.
Rollins BJ. Chemokines. Blood. 1997;90:909–928.
23. Collins T. Endothelial nuclear factor-kappa B and the initiation of the atherosclerotic lesion. Lab Invest. 1993;68:499–508.[Medline] [Order article via Infotrieve]
24.
Yla-Herttuala S, Lipton BA, Rosenfeld ME, et al.
Expression of monocyte chemoattractant protein 1 in
macrophage-rich areas of human and rabbit atherosclerotic
lesions. Proc Natl Acad Sci U S A. 1991;88:5252–5256.
25.
Usui M, Egashira K, Kitamoto S, et al. Pathogenic role
of oxidative stress in vascular angiotensin-converting
enzyme activation in long-term blockade of nitric oxide synthesis in
rats. Hypertension. 1999;34:546–551.
26.
Morishita R, Higaki J, Tomita N, et al. Application of
transcription factor "decoy" strategy as means of gene therapy and
study of gene expression in cardiovascular disease.
Circ Res. 1998;82:1023–1028.
27. Morishita R, Sugimoto T, Aoki M, et al. In vivo transfection of cis element "decoy" against nuclear factor-kappaB binding site prevents myocardial infarction. Nat Med. 1997;3:894–899.[Medline] [Order article via Infotrieve]
28. Sakanashi Y, Takeya M, Yoshimura T, et al. Kinetics of macrophage subpopulations and expression of monocyte chemoattractant protein-1 (MCP-1) in bleomycin-induced lung injury of rats studied by a novel monoclonal antibody against rat MCP-1. J Leukoc Biol. 1994;56:741–750.[Abstract]
29. Ueda A, Okuda K, Ohno S, et al. NF-kappa B and Sp1 regulate transcription of the human monocyte chemoattractant protein-1 gene. J Immunol. 1994;153:2052–2063.[Abstract]
30. Porreca E, Di Febbo C, Reale M, et al. Monocyte chemotactic protein 1 (MCP-1) is a mitogen for cultured rat vascular smooth muscle cells. J Vasc Res. 1997;34:58–65.[Medline] [Order article via Infotrieve]
31.
Weber KT. Extracellular matrix remodeling in heart
failure: a role for de novo angiotensin II generation.
Circulation. 1997;96:4065–4082.
This article has been cited by other articles:
 |
|
 |
|
  S. Kimura, K. Egashira, L. Chen, K. Nakano, E. Iwata, M. Miyagawa, H. Tsujimoto, K. Hara, R. Morishita, K. Sueishi, et al. Nanoparticle-Mediated Delivery of Nuclear Factor {kappa}B Decoy Into Lungs Ameliorates Monocrotaline-Induced Pulmonary Arterial Hypertension Hypertension, May 1, 2009; 53(5): 877 - 883. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. K. Upmacis, M. J. Crabtree, R. S. Deeb, H. Shen, P. B. Lane, L. E. S. Benguigui, N. Maeda, D. P. Hajjar, and S. S. Gross Profound biopterin oxidation and protein tyrosine nitration in tissues of ApoE-null mice on an atherogenic diet: contribution of inducible nitric oxide synthase Am J Physiol Heart Circ Physiol, November 1, 2007; 293(5): H2878 - H2887. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. Sawada, Y. Mitani, J. Maruyama, B. H. Jiang, Y. Ikeyama, F. A. Dida, H. Yamamoto, K. Imanaka-Yoshida, H. Shimpo, A. Mizoguchi, et al. A Nuclear Factor-{kappa}B Inhibitor Pyrrolidine Dithiocarbamate Ameliorates Pulmonary Hypertension in Rats Chest, October 1, 2007; 132(4): 1265 - 1274. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Ohtani, K. Egashira, K. Nakano, G. Zhao, K. Funakoshi, Y. Ihara, S. Kimura, R. Tominaga, R. Morishita, and K. Sunagawa Stent-Based Local Delivery of Nuclear Factor-{kappa}B Decoy Attenuates In-Stent Restenosis in Hypercholesterolemic Rabbits Circulation, December 19, 2006; 114(25): 2773 - 2779. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. Buffoli, O. Pechanova, S. Kojsova, R. Andriantsitohaina, L. Giugno, R. Bianchi, and R. Rezzani Provinol Prevents CsA-induced Nephrotoxicity by Reducing Reactive Oxygen Species, iNOS, and NF-kB Expression J. Histochem. Cytochem., December 1, 2005; 53(12): 1459 - 1468. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. P Marso, J. W Murphy, J. A House, D. M Safley, and W. S Harris Metabolic syndrome-mediated inflammation following elective percutaneous coronary intervention Diabetes and Vascular Disease Research, February 1, 2005; 2(1): 31 - 36. [Abstract] [PDF]
|
|
|
|
|
|
Q. Zhao, M. Ishibashi, K.-i. Hiasa, C. Tan, A. Takeshita, and K. Egashira Essential Role of Vascular Endothelial Growth Factor in Angiotensin II-Induced Vascular Inflammation and Remodeling Hypertension, September 1, 2004; 44(3): 264 - 270. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Dupuis, F. Soubrier, I. Brocheriou, S. Raoux, M. Haloui, L. Louedec, J.-B. Michel, and S. Nadaud Profiling of Aortic Smooth Muscle Cell Gene Expression in Response to Chronic Inhibition of Nitric Oxide Synthase in Rats Circulation, August 17, 2004; 110(7): 867 - 873. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Fiorucci, A. Mencarelli, A. Meneguzzi, A. Lechi, B. Renga, P. del Soldato, A. Morelli, and P. Minuz Co-administration of nitric oxide-aspirin (NCX-4016) and aspirin prevents platelet and monocyte activation and protects against gastric damage induced by aspirin in humans J. Am. Coll. Cardiol., August 4, 2004; 44(3): 635 - 641. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. L. Unthank, K. M. Sheridan, and M. C. Dalsing Collateral Growth in the Peripheral Circulation: A Review Vascular and Endovascular Surgery, July 1, 2004; 38(4): 291 - 313. [Abstract] [PDF]
|
|
|
|
 |
|
 H. F. Galley and N. R. Webster Physiology of the endothelium Br. J. Anaesth., July 1, 2004; 93(1): 105 - 113. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Ishibashi, K.-i. Hiasa, Q. Zhao, S. Inoue, K. Ohtani, S. Kitamoto, M. Tsuchihashi, T. Sugaya, I. F. Charo, S. Kura, et al. Critical Role of Monocyte Chemoattractant Protein-1 Receptor CCR2 on Monocytes in Hypertension-Induced Vascular Inflammation and Remodeling Circ. Res., May 14, 2004; 94(9): 1203 - 1210. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Kataoka, K. Egashira, M. Ishibashi, S. Inoue, W. Ni, K.-i. Hiasa, S. Kitamoto, M. Usui, and A. Takeshita Novel anti-inflammatory actions of amlodipine in a rat model of arteriosclerosis induced by long-term inhibition of nitric oxide synthesis Am J Physiol Heart Circ Physiol, February 1, 2004; 286(2): H768 - H774. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. A. Lemarie, B. Esposito, A. Tedgui, and S. Lehoux Pressure-Induced Vascular Activation of Nuclear Factor-{kappa}B: Role in Cell Survival Circ. Res., August 8, 2003; 93(3): 207 - 212. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Santos, V. I. Peinado, J. Ramirez, J. Morales-Blanhir, R. Bastos, J. Roca, R. Rodriguez-Roisin, and J. A. Barbera Enhanced Expression of Vascular Endothelial Growth Factor in Pulmonary Arteries of Smokers and Patients with Moderate Chronic Obstructive Pulmonary Disease Am. J. Respir. Crit. Care Med., May 1, 2003; 167(9): 1250 - 1256. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. Zauli, A. Pandolfi, A. Gonelli, R. Di Pietro, S. Guarnieri, G. Ciabattoni, R. Rana, M. Vitale, and P. Secchiero Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand (TRAIL) Sequentially Upregulates Nitric Oxide and Prostanoid Production in Primary Human Endothelial Cells Circ. Res., April 18, 2003; 92(7): 732 - 740. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Egashira Molecular Mechanisms Mediating Inflammation in Vascular Disease: Special Reference to Monocyte Chemoattractant Protein-1 Hypertension, March 1, 2003; 41(3): 834 - 841. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Ishibashi, K. Egashira, K.-i. Hiasa, S. Inoue, W. Ni, Q. Zhao, M. Usui, S. Kitamoto, T. Ichiki, and A. Takeshita Antiinflammatory and Antiarteriosclerotic Effects of Pioglitazone Hypertension, November 1, 2002; 40(5): 687 - 693. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Mori, K. Komori, T. Yamaoka, M. Tanii, C. Kataoka, A. Takeshita, M. Usui, K. Egashira, and K. Sugimachi Essential Role of Monocyte Chemoattractant Protein-1 in Development of Restenotic Changes (Neointimal Hyperplasia and Constrictive Remodeling) After Balloon Angioplasty in Hypercholesterolemic Rabbits Circulation, June 18, 2002; 105(24): 2905 - 2910. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Egashira, Q. Zhao, C. Kataoka, K. Ohtani, M. Usui, I. F. Charo, K.-i. Nishida, S. Inoue, M. Katoh, T. Ichiki, et al. Importance of Monocyte Chemoattractant Protein-1 Pathway in Neointimal Hyperplasia After Periarterial Injury in Mice and Monkeys Circ. Res., June 14, 2002; 90(11): 1167 - 1172. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
I. Bernatova, O. Pechanova, P. Babal, S. Kysela, S. Stvrtina, and R. Andriantsitohaina Wine polyphenols improve cardiovascular remodeling and vascular function in NO-deficient hypertension Am J Physiol Heart Circ Physiol, March 1, 2002; 282(3): H942 - H948. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Kataoka, K. Egashira, S. Inoue, M. Takemoto, W. Ni, M. Koyanagi, S. Kitamoto, M. Usui, K. Kaibuchi, H. Shimokawa, et al. Important Role of Rho-kinase in the Pathogenesis of Cardiovascular Inflammation and Remodeling Induced by Long-Term Blockade of Nitric Oxide Synthesis in Rats Hypertension, February 1, 2002; 39(2): 245 - 250. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. Z. Ammarguellat, P. O. Gannon, F. Amiri, and E. L. Schiffrin Fibrosis, Matrix Metalloproteinases, and Inflammation in the Heart of DOCA-Salt Hypertensive Rats: Role of ETA Receptors Hypertension, February 1, 2002; 39(2): 679 - 684. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Adamopoulos, J. T. Parissis, and D. Th. Kremastinos A glossary of circulating cytokines in chronic heart failure Eur J Heart Fail, October 1, 2001; 3(5): 517 - 526. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Tedgui and Z. Mallat Anti-Inflammatory Mechanisms in the Vascular Wall Circ. Res., May 11, 2001; 88(9): 877 - 887. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. C. Luft Workshop: Mechanisms and Cardiovascular Damage in Hypertension Hypertension, February 1, 2001; 37(2): 594 - 598. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C.-L. M. Cooke and S. T. Davidge Peroxynitrite increases iNOS through NF-kappa B and decreases prostacyclin synthase in endothelial cells Am J Physiol Cell Physiol, February 1, 2002; 282(2): C395 - C402. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Kubo-Inoue, K. Egashira, M. Usui, M. Takemoto, K. Ohtani, M. Katoh, H. Shimokawa, and A. Takeshita Long-term inhibition of nitric oxide synthesis increases arterial thrombogenecity in rat carotid artery Am J Physiol Heart Circ Physiol, April 1, 2002; 282(4): H1478 - H1484. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
O. Yokoseki, J.-i. Suzuki, H. Kitabayashi, N. Watanabe, Y. Wada, M. Aoki, R. Morishita, Y. Kaneda, T. Ogihara, H. Futamatsu, et al. cis Element Decoy Against Nuclear Factor-{kappa}B Attenuates Development of Experimental Autoimmune Myocarditis in Rats Circ. Res., November 9, 2001; 89(10): 899 - 906. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Q. Zhao, K. Egashira, S. Inoue, M. Usui, S. Kitamoto, W. Ni, M. Ishibashi, K.-i. Hiasa, T. Ichiki, M. Shibuya, et al. Vascular Endothelial Growth Factor Is Necessary in the Development of Arteriosclerosis by Recruiting/Activating Monocytes in a Rat Model of Long-Term Inhibition of Nitric Oxide Synthesis Circulation, March 5, 2002; 105(9): 1110 - 1115. [Abstract] [Full Text] [PDF]
|
|
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||