© 2001 American Heart Association, Inc.
Vascular Biology |
Endothelial NO Synthase Overexpression Inhibits Lesion Formation in Mouse Model of Vascular Remodeling
From the First Department of Internal Medicine (S.K., T.Y., M.O., Y.O., N.I., K.H., M.Y.) and the First Department of Pathology (H.A., Y.H., H.I.), Kobe University School of Medicine, Kobe, Japan.
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Abstract |
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Abstract—NO produced by endothelial NO synthase (eNOS) plays important roles in the regulation of vascular tone and structure. The purpose of this study was to clarify the role of eNOS-derived NO on vascular remodeling by use of eNOS-transgenic (eNOS-Tg) mice. The common carotid artery was ligated just proximal to the carotid bifurcation. Four weeks later, the proximal carotid artery of the ligation site was histologically examined. In this vascular remodeling model, the endothelium remains uninjured, but neointimal and medial thickening occurs in combination with a reduction in vascular diameter at the proximal portion of the ligation. At 4 weeks after ligation, the respective neointimal and medial areas in wild-type mice were 17 200±1100 and 24 300±1500 µm2, whereas both were reduced to 8000±1900 (P<0.01) and 18 400±700 µm2 (P<0.01) in eNOS-Tg mice (n=8). Total vascular area was not different between the 2 genotypes. NG-Nitro-L-arginine methyl ester treatment increased neointimal and medial areas to the same extent in both genotypes. Leukocyte infiltration was observed in the luminal side of the vessel, but the number of infiltrating cells was significantly attenuated in eNOS-Tg mice compared with wild-type mice. This reduction of leukocyte infiltration in eNOS-Tg mice was associated with reduced expressions of intracellular adhesion molecule-1 and vascular cellular adhesion molecule-1 on the endothelium. In conclusion, chronic eNOS overexpression in the endothelium reduced leukocyte infiltration and inhibited neointimal formation and medial thickening. Our data provide the evidence for the regulatory role of NO from the endothelium on vascular structure integrity.
Key Words: nitric oxide • endothelial nitric oxide synthase • transgenic mouse • vascular remodeling • intimal thickening
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Introduction |
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Vascular remodeling is an adaptive process that occurs in response to chronic changes in blood flow and other hemodynamic conditions, and a variety of vascular cellular responses, such as the growth and death of vascular smooth muscle cells and changes in extracellular matrix composition, are involved in the mechanisms.1 2 Endothelial cells are thought to play a cardinal role in sensing changes in mechanical and biochemical forces and to regulate vascular structure.3 One of the key molecules released by the endothelium is NO, synthesized by endothelial NO synthase (eNOS), which diffuses to underlying vascular smooth muscle cells or the lumen side and affects various cell functions. Over the past years, several studies have been performed involving the importance of endogenous eNOS-derived NO in vascular remodeling with the use of NO synthase (NOS) inhibitor–treated animals and eNOS-deficient mice.4 5 6 Furthermore, eNOS gene transfer to the balloon-injured arteries in animal experiments has been shown to inhibit neointimal formation.7 8 9 However, in those studies, eNOS was overexpressed mainly in vascular smooth muscle cells and/or the adventitia, not in the endothelium. On the other hand, no studies have ever tried to examine the effect of chronic overproduction of eNOS-derived NO from the endothelium on vascular remodeling. Recently, we generated transgenic (Tg) mice that overexpress eNOS in the endothelium. We reported that the eNOS-Tg mice exhibit hypotension with increased NO production.10 We also found that the basal eNOS activity in the vessels showed a 7-fold increase and that acetylcholine-induced NO production was elevated 3-fold in eNOS-Tg mice compared wild-type mice.10 11 In the present study, to clarify the effects of chronic NO overproduction on vascular remodeling, particularly on lesion formation, we examined, with the use of eNOS-Tg mice, the mouse carotid artery vascular remodeling model12 in which the endothelium remains uninjured.
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Methods |
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Animal Preparation.
We have generated Tg mice overexpressing the bovine eNOS gene in the endothelium (eNOS-Tg mice) by use of the preproendothelin-1 promoter.10 Heterozygous Tg mice and their littermate wild-type mice were used at 12 to 16 weeks of age. To inhibit NOS chronically, some of mice were provided water containing 1 mg/mL NG-nitro-L-arginine methyl ester (L-NAME). This dose of L-NAME increased blood pressure in both genotypes and canceled the difference in blood pressure, as reported previously.10 Several wild-type mice were fed 0.12 mg/mL hydralazine containing water to reduce blood pressure to the levels found in eNOS-Tg mice. L-NAME and hydralazine were both started 2 weeks before the carotid ligation and were continued throughout the experiment. Moreover, some mice were provided chow containing 0.05% aspirin. The chow was started 1 week before the ligation and was continued for 3 weeks. All animal experiments were conducted according to the Guidelines for Animal Experiments at Kobe University School of Medicine.
Vascular Remodeling Model in Mouse Carotid
Artery
We used a mouse model of vascular remodeling by
carotid ligation, which has been reported
previously.12 After
anesthesia by intraperitoneal injection
of pentobarbital sodium, left carotid arteries were dissected and
ligated completely near the carotid bifurcation. Groups of animals were
killed at 3, 7, 14, and 28 days after ligation of the carotid artery.
All animals received 2 subcutaneous injections of the thymidine
analogue bromodeoxyuridine (BrdU, 2.5 mg per injection, Sigma Chemical
Co) 24 and 6 hours before euthanasia. All mice were fixed for 5 minutes
under their mean pressures (95 mm Hg [wild type] and 80
mm Hg [Tg]) with 4% paraformaldehyde in PBS. After
excision of the left and right carotid arteries, the vessels were
immersion-fixed in 4% paraformaldehyde in PBS and
embedded in paraffin or in OCT compounds (Miles Laboratories),
and serial sections were cut for analysis by hematoxylin-eosin
staining and immunostaining. Serial sections collected
every 50-µm interval (0.5- to 3.0-mm segments proximal to the
ligation site) were used for morphological analysis. Similar
segments of the right carotid artery were analyzed for the
control condition.
Morphological Analysis
Morphological analysis was carried out on
left (ligated side) and right (nonligated side) carotid arteries
harvested 4 weeks after ligation according to the method originally
described by Kumar et al.13
All serial sections collected from the 0.5- to 3.0-mm segments proximal
to the ligation site were divided into 5 nearly equal groups, and 2
sections were picked from each group. Thus, in total, 10 sections were
measured, and data for each mouse were shown as the average of 10
sections. This procedure was performed to ensure that the entire length
of the arterial sample was used in the analysis,
because the thickness of the lesion varied along the length of the
artery. Digitized images of these vessels were analyzed with
NIH 1.58 Imaging Software. The circumference length of the lumen,
internal elastic lamina (IEL), and external elastic lamina (EEL) were
determined by tracing along the luminal surface, IEL, and EEL,
respectively. Under the assumption that the structures were circular,
we calculated the areas enclosed by the EEL, IEL, and lumen. The area
defined by the EEL was used as the total vascular area. The medial area
was calculated by subtracting the area defined by the IEL from the
total vascular area, and the intimal area was determined by subtracting
the lumen area from the area defined by the IEL.
Immunohistochemistry and
Immunoblotting
Immunohistochemical staining for eNOS was obtained
with a rabbit polyclonal eNOS antibody (1:50 dilution, Transduction
Laboratories) with the use of frozen sections as previously
reported.10 14 At 2
weeks after ligation, protein was extracted from the ligated and
nonligated carotid arteries with the use of 10 mice in each group. The
membrane fraction of the protein was separated by
ultracentrifugation and immunoblotted.
Immunoblotting of eNOS was performed as previously
described.10 Briefly, 100
µg of protein samples from the membrane fraction was separated on a
7.5% SDS-polyacrylamide gel under reducing conditions. After
electrophoresis, the protein was transferred to a nitrocellulose
membrane. Blots were then incubated in a blocking buffer consisted of
5% nonfat dry milk and incubated overnight at 4°C with a rabbit
polyclonal eNOS antibody (1:200 dilution) described above.
Immunoreactive bands were visualized by use of an ECL detection kit
(Amersham). We also assessed the expression of inducible NOS (iNOS) by
histochemistry with the use of a rabbit polyclonal anti-iNOS antibody
(1:500 dilution, Santa Cruz Biotechnology). The vascular cell
replication indices in the intima and media were determined by staining
with a rat monoclonal antibody against BrdU (1:200 dilution, Harlan
SERA-LAB). The numbers of total and stained nuclei were counted, and
the percentage of the BrdU labeling was
calculated.12 The presence of
inflammatory cells was determined by using a rat monoclonal antibody
against the mouse CD45, a leukocyte common antigen (1:250 dilution,
Pharmingen).13 The
expressions of intracellular adhesion molecule-1 (ICAM-1) and vascular
cellular adhesion molecule-1 (VCAM-1) were evaluated by
immunostaining with use of a mouse monoclonal
anti–ICAM-1 antibody (1:100 dilution, R&D Systems) and a goat
polyclonal anti–VCAM-1 antibody (1:200 dilution, Santa Cruz
Biotechnology), respectively. Biotinylated secondary antibodies and
streptavidin–horseradish peroxidase (DAKOPATTS) with
3,3'-diaminobenzidine as the color substrate were used for all staining
runs. At least 3 sections were analyzed per animal for these
factors. The extents of ICAM-1 and VCAM-1 expressions were
semiquantitatively evaluated by scoring as follows: grade 0, negative
stain; grade 1, variable or weak stain; and grade 2, moderately or
strongly positive stain.15
The sections were blindly graded by 2 independent senior
pathologists.
Statistical Analysis
Data are presented as mean±SEM. An unpaired
Student t test or Mann-Whitney
U test was used to detect
significant differences when 2 groups were compared. Statistical
differences among group means were determined by 1-way ANOVA with
repeated measures, followed by a post hoc comparison. A value of
P<0.05 was considered to be
statistically significant.
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Results |
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Comparison of Morphological Changes in eNOS-Tg and Wild-Type Mice
In this vascular remodeling model, the endothelium remained intact, but the luminal area was reduced through a combination of decreased vessel diameter, neointimal formation, and medial thickening at the proximal portion of the ligation.12 In nonligated side carotid arteries, there were no significant differences in luminal, intimal, medial, and total vascular areas between the 2 genotypes (Table 1
). At 4 weeks after ligation, the
neointimal and medial areas were significantly reduced in
eNOS-Tg mice compared with wild-type mice
(Figure 1B, 1C, and 1E and
Table 1). The total vascular area as measured by the area
defined by EEL was not different between the two genotypes,
whereas the luminal area was markedly larger in eNOS-Tg mice
(Figure 1E and
Table 1).
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Hydralazine treatment reduced blood pressure by
19±4 mm Hg in wild-type mice to levels comparable to those in
eNOS-Tg mice, whereas it did not influence the intimal (17 000±5200
µm2) and medial (29 700±2300
µm2) thickening (n=5 treated wild-type
mice, P=NS versus nontreated wild-type mice;
Table 1
). L-NAME treatment increased the
neointimal and medial areas in wild-type and eNOS-Tg mice
(Figure 1D and
Table 1). There were no differences in those values between
the 2 genotypes.
Morphology and Immunohistochemistry
The presence of endothelial cells on
the luminal surface was verified by immunostaining for
von Willebrand factor at various time points after ligation. In
accord with a study of Kumar et
al,12 we noted in the
present study an occasional clot formation only in the immediate
vicinity of the ligature, and we did not find clot formation at the
portion >1 mm proximal to the ligation site. We also confirmed
that aspirin treatment did not alter the process of lesion formation in
this model.
Immunohistochemistry and immunoblotting
revealed that eNOS expression at the endothelium was
increased in eNOS-Tg mice in nonligated carotid arteries
(Figure 2A through 2E). Carotid ligation dramatically
decreased eNOS expression at the endothelium proximal
to the ligated point in wild-type mice, whereas its expression was
relatively maintained in eNOS-Tg mice
(Figure 2A through 2D).
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Inflammatory cells were present near the luminal surface
at 3 and 7 days after ligation. At 3 days after ligation, the number of
CD45-positive cells around the luminal surface (total counts of 5
sections for each mouse) was significantly reduced in eNOS-Tg mice
compared with wild-type mice (136.2±26.2 versus 308.8±34.4,
respectively; P<0.01;
Figure 3). Inflammatory cells were also found in the
adventitia and the outer layer of the media within 2 weeks after
ligation. Although the mechanisms of the inflammatory changes in the
adventitia and the outer layer of the media are not yet explored, they
may, at least in part, be related to the surgical procedure. Those
inflammatory cells were diminished, and only a few cells were seen in
the adventitia at 4 weeks after ligation
(Figure 1B and 1C). In association with these inflammatory
responses, iNOS expression was detected mainly in the adventitia,
especially in the infiltrated inflammatory cells, and the expression
seemed to be not different between wild-type and eNOS-Tg mice at 1 week
after ligation
(Figure 2F and 2G).
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To clarify the mechanism of reduced leukocyte infiltration
from the luminal side, we examined expressions of adhesion molecules by
immunohistochemistry. Carotid ligation markedly increased ICAM-1
expression in wild-type mice at 3 and 7 days after ligation, whereas
the expression in eNOS-Tg mice was still obscure and reduced compared
with that in wild-type mice
(Figure 4A and 4B and
Table 2). Likewise, VCAM-1 expression was significantly
reduced in eNOS-Tg mice compared with wild-type mice at 3 and 7 days
after ligation
(Figure 4C and 4D and
Table 2). In contrast, expressions of ICAM-1 and VCAM-1 in
the contralateral right carotid artery were obscure, and they did not
change significantly after ligation of the left carotid in both
genotypes.
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As described above, carotid ligation induced
neointimal and medial thickening. To examine whether the
increase in wall thickness after blood flow cessation was due to
hyperplasia or hypertrophy of smooth muscle cells, we
assessed the incorporation of BrdU into carotid vessels. The percentage
of BrdU-positive cells was significantly reduced in eNOS-Tg mice
compared with wild-type mice at 1 and 2 weeks after the carotid
ligation
(Figure 5).
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Discussion |
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The present study using eNOS-Tg mice demonstrated that NO overproduction from the endothelium inhibits the vascular lesion formation in the carotid artery vascular remodeling model. In eNOS-Tg mice, the reduced lesion formation was associated with attenuated inflammatory cell infiltration. L-NAME treatment aggravated the lesion formation in wild-type and eNOS-Tg mice and resulted in a similar extent of lesion formation between them.
Endothelium-derived NO is thought to play an important role in vascular remodeling. A number of studies using experimental balloon angioplasty demonstrated the protective role of NO derived from eNOS on vascular remodeling characterized by neointimal formation with the use of eNOS gene transfer or the administration of L-arginine, a substrate for NO.7 8 9 16 17 However, in the balloon angioplasty–induced vascular remodeling model, the endothelium is injured, and the role of NO produced from the endothelium cannot be fully elucidated. Therefore, the use of the vascular remodeling model in which the endothelium remains intact during the lesion formation process would be required to elucidate directly the role of NO from the endothelium on remodeling.10 Flow-induced vascular remodeling is a typical example of such a vascular remodeling model. Vessels undergo vascular remodeling characterized by intimal hyperplasia and increased vessel diameter in response to blood flow increase.18 19 In this type of vascular remodeling, the endothelium plays a critical role, and NOS inhibitors attenuate the remodeling process. On the other hand, vessel diameter decreases when blood flow is reduced.20 It has been demonstrated that cessation of blood flow by complete ligation of the vessel near the carotid bifurcation induces a combination of intimal and medial thickening together with decreased vessel diameter, which results in a reduction in the luminal area.12 Although the mechanisms of vascular remodeling in this model are not fully clarified, it is proposed that decreased shear stress due to cessation of flow results in shrinkage in the luminal area and thickening in the neointima and media of the vessel.12 It is also suggested that the inflammatory cell infiltration is implicated in the initiation of these process.13 In this remodeling model, the contralateral carotid artery is also shown to undergo a remodeling process due to increased blood flow; therefore, the extent of remodeling in the ligated artery may be overestimated. However, this vascular remodeling model, in which the endothelium is not removed and is kept intact, enables us to investigate the role of endogenous endothelium-derived NO on vascular remodeling.
The present finding is in accordance with the recent reports on vascular remodeling with the use of eNOS-deficient mice.5 6 In eNOS-deficient mice, deficiency of NO from the endothelium was shown to aggravate neointimal hyperplasia in 2 different types of vascular remodeling models with preserved endothelium. Rudic et al5 showed that eNOS-deficient mice exhibited acceleration of vascular remodeling in the common carotid artery after ligation of the ipsilateral external carotid artery. Likewise, Moroi et al6 reported that the intimal hyperplasia in the femoral artery caused by cuff placement was exaggerated in eNOS-deficient mice. Before and after carotid ligation, we found increased eNOS expression on the endothelium in eNOS-Tg mice. It is unclear why eNOS expression was preserved despite the reduction of shear stress in eNOS-Tg mice after ligation. However, it may be related to the use of the preproendothelin-1 promoter for the transgene, inasmuch as the expression of preproendothelin-1 is shown to increase by reduction of shear stress. The inhibitory effects in eNOS-Tg mice were abolished by L-NAME treatment. The vascular effects induced by L-NAME may not be entirely due to the inhibition of endogenous NO, but our finding is in agreement with previous reports demonstrating the aggravation of balloon angioplasty–induced vascular remodeling by L-NAME treatment in animal models.21 Thus, the attenuation of vascular remodeling in eNOS-Tg mice is caused by overproduced NO.
It was implicated that leukocyte recruitment plays critical roles in the present vascular remodeling model.12 13 The reduced blood flow results in activation of leukocyte- endothelial adhesion and induces subsequent influx of inflammatory cells. Indeed, we found the prominent leukocyte infiltration mainly in the luminal side of intimal hyperplasia in wild-type mice. Leukocyte-endothelial adhesion plays a cardinal role in the initiation of leukocyte infiltration. In the present study, we found upregulation of VCAM-1, an endothelial adhesion molecule, at the remodeling site by cessation of flow. Walpola et al22 also demonstrated the upregulation of VCAM-1 by a reduction of blood flow in a rabbit model. The importance of the adhesion molecule in lesion formation in this model was revealed by Kumar et al.13 They showed the reduced infiltration of inflammatory cells associated with intimal thickening in P-selectin knockout mice. In the present study, in eNOS-Tg mice, the number of leukocytes present in the developing intima and near the luminal surface was markedly reduced compared with that in wild-type mice. Most important, we found that the expressions of not only VCAM-1 but also ICAM-1 after carotid ligation were significantly reduced in eNOS-Tg mice compared with wild-type mice. It was revealed in studies in vitro that NO reduces lipopolysaccharide- or cytokine-induced expressions of adhesion molecules in endothelial cells.23 24 Using these eNOS-Tg mice, we have demonstrated that overproduction of NO from pulmonary vascular endothelial cells reduced lipopolysaccharide-induced upregulation of adhesion molecules in the lungs.25 Thus, it seems that suppression of leukocyte infiltration resulting from the inhibition of adhesion molecule expressions by overproduced NO from the endothelium is involved in the mechanisms of the reduced vascular remodeling in eNOS-Tg mice, although other mechanisms may also be implicated.
In the present study, we demonstrated that the percentage of BrdU-positive cells was reduced in eNOS-Tg mice compared with wild-type mice. Thus, vascular smooth muscle cell proliferation was reduced in eNOS-Tg mice. Because the inflammatory cell infiltration was attenuated in eNOS-Tg mice, the production of various growth stimuli on smooth muscle cells from those mice was likely to be reduced. It is also possible that the overproduced NO directly inhibited the proliferation of vascular smooth muscle cells. Studies in vitro showed that NO donors and cGMP-elevating agents inhibit vascular smooth muscle cell proliferation.26 27 Although the mechanisms are still not fully elucidated, recent reports have implicated that NO affects some cell cycle–associated proteins, such as cyclin A.28
In the present study, overexpression of eNOS in the endothelium reduced neointimal and medial thickening but did not affect the reduction in vessel diameter. The mechanisms of vascular constriction in this model are unclear, and so far, only the participation of the cytoskeleton29 and the role of iNOS detected in the adventitia and the outer layer have been suggested.30 Further studies are needed to clarify the mechanisms, particularly the role of iNOS-derived NO, of the reduction in vascular diameter associated with the cessation of blood flow in this model.
In conclusion, the present study suggests that NO produced from endothelial cells inhibits neointimal formation and medial thickening of the carotid artery induced by the cessation of blood flow in mice. Although the mechanisms of the vascular remodeling process used in the present study are not fully clarified and factors other than NO are likely involved in them, our results provide direct evidence of the regulatory role of NO from the endothelium on vascular structure integrity.
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Acknowledgments |
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This work was supported by Grants-in-Aid for Scientific Research on Priority Areas and Grants-in-Aid for Scientific Research from the Ministry of Education, Science, Sports, and Culture, Japan. We gratefully appreciate Kiyoko Matsui for secretarial assistance and technical support for animal care.
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Footnotes |
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Reprint requests to Seinosuke Kawashima, MD, PhD, First Department of Internal Medicine, Kobe University School of Medicine, 7-5-1 Kusunoki-cho, Chuo-ku, Kobe, 650-0017, Japan.
Received December 28, 1999; accepted November 27, 2000.
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References |
|---|
|
TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
-
Gibbons GH,
Dzau VJ. The emerging concept of vascular remodeling.
N Engl J Med. 1994;330:1431–1438.
[Free Full Text] - Mulvany MJ, Baumbach GL, Aalkjaer C, Heagerty AM, Korsgaard N, Schiffrin EL, Heistad DD. Vascular remodeling. Hypertension. 1996;28:505–506.
-
Langille BL,
O’Donnell F. Reductions in arterial diameter produced by
chronic decreases in blood flow are
endothelium-dependent.
Science. 1986;231:405–407.
[Abstract/Free Full Text] - Takemoto M, Egashira K, Usui M, Numaguchi K, Tomita H, Tsutsui H, Shimokawa H, Sueishi K, Takeshita A. 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]
- Rudic DR, Shesely EG, Maeda N, Smithies O, Segal SS, Sessa WC. 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]
- Moroi M, Zhang L, Yasuda T, Virmani R, Gold HK, Fishman MC, Huang PL. 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]
-
von der Leyen HE,
Gibbons GH, Morishita R, Lewis NP, Zhang L, Nakajima M, Kaneda Y, Cooke
JP, Dzau VJ. 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.
[Abstract/Free Full Text] -
Varenne O, Pislaru
S, Gillijns H, Van Pelt D, Gerard RD, Zoldhelyi P, Van de Werf F,
Collen D, Janssens SP. Local adenovirus-mediated transfer of human
endothelial nitric oxide synthase reduces luminal
narrowing after coronary angioplasty in pigs.
Circulation.. 1998;98:919–926.
[Abstract/Free Full Text] - Shears LL II, Kibbe MR, Murdock AD, Billar TR, Lizonava A, Kovesdi I, Watkins SC, Tzenng E. Efficient inhibition of intimal hyperplasia by adenovirus-mediated inducible nitric oxide synthase gene transfer to rats and pigs in vivo. J Am Coll Surg. 1998;187:295–306.[Medline] [Order article via Infotrieve]
- Ohashi Y, Kawashima S, Hirata K, Yamashita T, Ishida T, Inoue N, Sakoda T, Kurihara H, Yazaki Y, Yokoyama M. Hypotension and reduced nitric oxide-elicited vasorelaxation in transgenic mice overexpressing endothelial nitric oxide synthase. J Clin Invest. 1998;102:2061–2071.[Medline] [Order article via Infotrieve]
-
Yamashita T,
Kawashima S, Ohashi Y, Ozaki M, Rikitake Y, Inoue N, Hirata K, Akita H,
Yokoyama M. Mechanisms of reduced NO/cGMP-mediated vasorelaxations in
transgenic mice overexpressing endothelial nitric oxide
synthase. Hypertension. 2000;36:97–102.
[Abstract/Free Full Text] -
Kumar A, Lindner
V. Remodeling with neointima formation in the mouse carotid
artery after cessation of blood flow.
Arterioscler Thromb Vasc Biol. 1997;17:2238–2244.
[Abstract/Free Full Text] -
Kumar A, Hoover
JL, Simmons CA, Lindner V, Shebuski RJ. Remodeling and
neointimal formation in the carotid artery of normal and
P-selectin-deficient mice.
Circulation. 1997;96:4333–4342.
[Abstract/Free Full Text] - Kanazawa K, Kawashima S, Mikami S, Miwa Y, Hirata K, Suematsu M, Hayashi Y, Itoh H, Yokoyama M. Endothelial constitutive nitric oxide synthase protein and mRNA increased in rabbit atherosclerotic aorta despite impaired endothelium-dependent vascular relaxation. Am J Pathol. 1996;148:1949–1956.[Abstract]
-
Azumi H, Nobutaka
I, Takeshita S, Rikitake Y, Kawashima S, Hayashi Y, Itoh H, Yakoyama M.
Expression of NADH/NADPH oxidase p22phox in human coronary
arteries. Circulation. 1999;100:1494–1498.
[Abstract/Free Full Text] -
Bosmans JM, Vrints
CJ, Kockx MM, Bult H, Cromheeke KMC, Herman AG. Continuous perivascular
L-arginine delivery increases total vessel area and reduces
neointimal thickening after experimental balloon dilation.
Arterioscler Thromb Vasc Biol. 1999;19:767–776.
[Abstract/Free Full Text] -
Tarry WC, Makhoul
RG. L-Arginine improves endothelium-dependent
vasorelaxation and reduces intimal hyperplasia after balloon
angioplasty. Arterioscler Thromb Vasc
Biol. 1994;14:938–944.
[Abstract/Free Full Text] - Rudic RD, Sessa WC. Human genetics ’99: the cardiovascular system: nitric oxide in endothelial dysfunction and vascular remodeling: clinical correlates and experimental links. Am J Hum Genet. 1999;64:673–677.[Medline] [Order article via Infotrieve]
-
Tronc F, Wassef M,
Esposito B, Henrion D, Glacov S, Tedgui A. Role of NO in flow-induced
remodeling of the rabbit common carotid artery.
Arterioscler Thromb Vasc Biol. 1996;16:1256–1262.
[Abstract/Free Full Text] -
Langille BL,
Bendeck MO, Keeley FW. Adaptation of carotid arteries of young and
mature rabbits to reduced carotid blood flow.
Am J Physiol. 1989;256:H931–H939.
[Abstract/Free Full Text] -
Le Tourneau T, Van
Belle E, Corseaux D, Vallet B, Lebuffe G, Dupuis B, Lablanche J-M,
McFadden E, Bauters C, Bertrand ME. Role of nitric oxide in
restenosis after experimental balloon angioplasty in the
hypercholesterolemic rabbit: effects on
neointimal hyperplasia and vascular remodeling.
J Am Coll Cardiol. 1999;33:876–882.
[Abstract/Free Full Text] -
Walpola PL,
Gotlieb AI, Cybulsky MI, Langille BL. Expression of ICAM-1 and VCAM-1
and monocyte adherence in arteries exposed to altered shear stress.
Arterioscler Thromb Vasc Biol. 1995;15:2–10.
[Abstract/Free Full Text] - Caterina RD, Libby P, Peng H-B, Thannickal VJ, Rajavashisth TB, Gimbrone MA, Shin WS, Liao JK. Nitric oxide decrease cytokine-induced endothelial activation. J Clin Invest. 1995;96:60–68.
-
Qian H-S,
Neplioueva V, Shetty GA, Channon KM, George SE. Nitric oxide synthase
gene therapy rapidly reduces adhesion molecule expression and
inflammatory cell infiltration in carotid arteries of
cholesterol-fed rabbits.
Circulation. 1999;99:2979–2982.
[Abstract/Free Full Text] -
Yamashita T,
Kawashima S, Ohashi Y, Ozaki M, Ueyama T, Ishida T, Inoue N, Hirata K,
Akita H, Yokoyama M. Resistance to endotoxin shock in transgenic mice
overexpressing endothelial nitric oxide synthase.
Circulation. 2000;101:931–937.
[Abstract/Free Full Text] - Garg UC, Hassid A. Nitric oxide-generating vasodilators and 8-bromo-cyclic guanosine monophosphate inhibit mitogenesis and proliferation of cultured rat vascular smooth muscle cells. J Clin Invest. 1989;83:1774–1777.
-
Kolpakov V, Gordon
D, Kulik TJ. Nitric oxide-generating compounds inhibit total protein
and collagen synthesis in cultured vascular smooth muscle cells.
Circ Res. 1995;76:305–309.
[Abstract/Free Full Text] -
Guo K, Andres V,
Walsh K. Nitric oxide–induced downregulation of cdk2 activity and
cyclin A gene transcription in vascular smooth muscle cells.
Circulation. 1998;97:2066–2072.
[Abstract/Free Full Text] -
Schiffers PMH,
Henrion D, Boulanger CM, Colucci-Guyon E, Langa-Vuves F, van Essen H,
Fazzi GE, Lévy BI, De Mey JGR. Altered flow–induced
arterial remodeling in vimentin-deficient mice.
Arterioscler Thromb Vasc Biol. 2000;20:611–666.
[Abstract/Free Full Text] - Yogo K, Shimokawa H, Kandabashi T, Miyata K. Different vasculoprotective roles of nitric oxide synthase isoforms in the vascular lesion formation in mice. Circulation. 1999;100(suppl I):I-114. Abstract.
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 J. A. Vita, M. Holbrook, J. Palmisano, S. M. Shenouda, W. B. Chung, N. M. Hamburg, B. R. Eskenazi, L. Joseph, and O. M. Shapira Flow-Induced Arterial Remodeling Relates to Endothelial Function in the Human Forearm Circulation, June 17, 2008; 117(24): 3126 - 3133. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Sasaki, T. Yamashita, T. Takaya, M. Shinohara, R. Shiraki, M. Takeda, N. Emoto, A. Fukatsu, T. Hayashi, K. Ikemoto, et al. Augmentation of Vascular Remodeling by Uncoupled Endothelial Nitric Oxide Synthase in a Mouse Model of Diabetes Mellitus Arterioscler. Thromb. Vasc. Biol., June 1, 2008; 28(6): 1068 - 1076. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 W.-W. Lin, Y.-C. Lin, T.-Y. Chang, S.-H. Tsai, H.-C. Ho, Y.-T. Chen, and V. C. Yang Caveolin-1 Expression Is Associated with Plaque Formation in Hypercholesterolemic Rabbits J. Histochem. Cytochem., August 1, 2006; 54(8): 897 - 904. [Abstract] [Full Text] [PDF]
|
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|
|
 |
|
 J. K. Bendall, N. J. Alp, N. Warrick, S. Cai, D. Adlam, K. Rockett, M. Yokoyama, S. Kawashima, and K. M. Channon Stoichiometric Relationships Between Endothelial Tetrahydrobiopterin, Endothelial NO Synthase (eNOS) Activity, and eNOS Coupling in Vivo: Insights From Transgenic Mice With Endothelial-Targeted GTP Cyclohydrolase 1 and eNOS Overexpression Circ. Res., October 28, 2005; 97(9): 864 - 871. [Abstract] [Full Text] [PDF]
|
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|
 |
|
 Y. Rikitake, H.-H. Kim, Z. Huang, M. Seto, K. Yano, T. Asano, M. A. Moskowitz, and J. K. Liao Inhibition of Rho Kinase (ROCK) Leads to Increased Cerebral Blood Flow and Stroke Protection Stroke, October 1, 2005; 36(10): 2251 - 2257. [Abstract] [Full Text] [PDF]
|
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 O. W.H. van der Heijden, Y. P.G. Essers, G. Fazzi, L. L.H. Peeters, J. G.R. De Mey, and G. J.J.M. van Eys Uterine Artery Remodeling and Reproductive Performance Are Impaired in Endothelial Nitric Oxide Synthase-Deficient Mice Biol Reprod, May 1, 2005; 72(5): 1161 - 1168. [Abstract] [Full Text] [PDF]
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 E. K. Walsh, H. Huang, Z. Wang, J. Williams, R. de Crom, R. van Haperen, C. I. Thompson, D. J. Lefer, and T. H. Hintze Control of myocardial oxygen consumption in transgenic mice overexpressing vascular eNOS Am J Physiol Heart Circ Physiol, November 1, 2004; 287(5): H2115 - H2121. [Abstract] [Full Text] [PDF]
|
|
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S. Kawashima and M. Yokoyama Dysfunction of Endothelial Nitric Oxide Synthase and Atherosclerosis Arterioscler. Thromb. Vasc. Biol., June 1, 2004; 24(6): 998 - 1005. [Abstract] [Full Text] [PDF]
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P. G. Frank, H. Lee, D. S. Park, N. N. Tandon, P. E. Scherer, and M. P. Lisanti Genetic Ablation of Caveolin-1 Confers Protection Against Atherosclerosis Arterioscler. Thromb. Vasc. Biol., January 1, 2004; 24(1): 98 - 105. [Abstract] [Full Text] [PDF]
|
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S. P. Jones, J. J. M. Greer, A. K. Kakkar, P. D. Ware, R. H. Turnage, M. Hicks, R. van Haperen, R. de Crom, S. Kawashima, M. Yokoyama, et al. Endothelial nitric oxide synthase overexpression attenuates myocardial reperfusion injury Am J Physiol Heart Circ Physiol, January 1, 2004; 286(1): H276 - H282. [Abstract] [Full Text] [PDF]
|
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T. Aizawa, H. Wei, J. M. Miano, J.-i. Abe, B. C. Berk, and C. Yan Role of Phosphodiesterase 3 in NO/cGMP-Mediated Antiinflammatory Effects in Vascular Smooth Muscle Cells Circ. Res., September 5, 2003; 93(5): 406 - 413. [Abstract] [Full Text] [PDF]
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D. L. Myers, K. J. Harmon, V. Lindner, and L. Liaw Alterations of Arterial Physiology in Osteopontin-Null Mice Arterioscler. Thromb. Vasc. Biol., June 1, 2003; 23(6): 1021 - 1028. [Abstract] [Full Text] [PDF]
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C. Yan, D. Kim, T. Aizawa, and B. C. Berk Functional Interplay Between Angiotensin II and Nitric Oxide: Cyclic GMP as a Key Mediator Arterioscler. Thromb. Vasc. Biol., January 1, 2003; 23(1): 26 - 36. [Abstract] [Full Text] [PDF]
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 M. Ozaki, S. Kawashima, T. Hirase, T. Yamashita, M. Namiki, N. Inoue, K.-i. Hirata, and M. Yokoyama Overexpression of Endothelial Nitric Oxide Synthase in Endothelial Cells Is Protective against Ischemia-Reperfusion Injury in Mouse Skeletal Muscle Am. J. Pathol., April 1, 2002; 160(4): 1335 - 1344. [Abstract] [Full Text] [PDF]
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