(Circulation. 2001;103:423.)
© 2001 American Heart Association, Inc.
Basic Science Reports |
Estradiol Accelerates Reendothelialization in Mouse Carotid Artery Through Estrogen Receptor-
but Not Estrogen Receptor-ß
From INSERM U397, Institut L. Bugnard, CHU Rangueil, Toulouse (L.B., F.B., J.F.A.), and Institut de Génétique et de Biologie Moléculaire et Cellulaire, CNRS/INSERM/ULP/Collège de France, Illkirch, CU de Strasbourg (A.K., S.D., P.C.), France.
Correspondence to J.F. Arnal, INSERM U397, Institut L. Bugnard, CHU Rangueil, 31403 Toulouse, France. E-mail arnal{at}rangueil.inserm.fr
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Abstract |
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 Top Abstract IntroductionMethodsResultsDiscussionReferences |
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Background—The atheroprotective effect of 17ß-estradiol (E2) has been suggested in women and clearly demonstrated in animals through both an effect on lipid metabolism and a direct effect on the cells of the arterial wall. It has been shown, for example, that E2 promotes endothelium-dependent relaxation and accelerates reendothelialization in rats. Similar studies have been undertaken in mice to appreciate the molecular mechanism of this process.
Methods and Results—We
report here a model of electric carotid injury adapted from that
described by Carmeliet et al (1997) that allows us to precisely
evaluate the reendothelialization process. We demonstrate that
E2 accelerates endothelial regeneration in
castrated female wild-type mice. In ovariectomized transgenic mice in
which either the estrogen receptor (ER)-
or ERß gene has been
disrupted, E2 accelerated reendothelialization
in female ERß knockout mice, whereas this effect was abolished in
female ER knockout mice.
Conclusions—This study
demonstrates that ER but not ERß mediates the beneficial effect of
E2 on reendothelialization and potentially the
prevention of
atherosclerosis.
Key Words: hormones • receptors • arteries • endothelium
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Introduction |
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TopAbstractIntroductionMethodsResultsDiscussionReferences |
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Epidemiological and observational studies1 2 support a vasculoprotective effect of estrogens. This is traditionally thought to be due to potentially favorable changes in blood lipids and lipoproteins,3 but a number of experimental studies4 5 6 7 8 9 strongly suggest a direct effect on the vascular system. Among the cell types present in the normal and atherosclerotic vessel wall, endothelium represents both a key actor in the pathophysiology of atherosclerosis and an important target for 17ß-estradiol (E2).
Twenty years of intensive experimental work has confirmed the crucial role of endothelium in the physiology of circulation. The endothelium is uniquely positioned at the interface between the blood and the vessel wall. As such, it performs multiple functions: it is involved in the regulation of coagulation, leukocyte adhesion in inflammation, vessel tone, and vascular smooth muscle growth and also acts as a barrier to transvascular flux of liquids and solutes. In addition, endothelial abnormalities appear to be central to the pathogenesis of atherosclerosis.10 11 The importance of endothelial integrity was initially demonstrated by the facilitation of atherosclerotic lesion development in hypercholesterolemic animal models after removal of endothelium. Although subsequent studies demonstrated the presence of endothelium overlying the lesions, repeated mechanical, hemodynamic, and/or immunological injury probably contributes to morphological and functional alterations of the endothelium as well as its senescence. Telomere length is decreased in cells of the arterial tree subject to hemodynamic stress and prone to atherosclerosis, demonstrating an accelerated cell turnover at these sites.12 Recent experimental work has demonstrated that low shear stress, which in some respects mimics turbulent blood flow, favors apoptosis of cultured endothelial cells.13 14 Altogether, these observations suggest that the capacity of endothelium to maintain a cell monolayer could be of crucial importance in the prevention of atherosclerosis and its complications.
Several models of arterial injury have been developed
to study endothelial regrowth. Injury of an artery by passage of a
balloon catheter causes endothelial denudation and medial damage and
produces both an endothelial and a smooth muscle cell proliferative
response in nonhuman primates, swine, dogs, rabbits, and
rats.15 In such models,
E2 was reported to promote endothelial regrowth
in castrated female rats.16
The effect of E2 has long been thought to be
mediated through activation of the estrogen receptor (ER)-. ER is
a ligand-dependent transcriptional
activator17 that modulates
gene expression in target cells not only in reproductive tissues but
also in bone and
vessels.18 19
Moreover, ERß, recently discovered and cloned from rat
prostate,20 was found to be
expressed in many other tissues, including injured
arteries.21 22
Evaluation of the respective roles of ER and ERß in the
reendothelialization process requires a reliable model of mouse
arterial injury in which the extent of endothelial regrowth can be
quantified. Removal of the endothelium in the mouse carotid artery with
a flexible wire was described in
199323 and allowed the
arterial smooth muscle proliferation and/or neointima formation in
various transgenic mice to be evaluated. This model of endovascular
carotid injury suggested that ER was not involved in one aspect of
the protective effect of E2, ie, the prevention
of medial smooth muscle
proliferation.24 However,
the specific evaluation of arterial reendothelialization appears to be
difficult with this
technique.23 Recently,
Carmeliet et al25 described
a novel model of electric injury to the mouse femoral artery that
consisted of destroying endothelial, smooth muscle, and adventitial
cells and allowing subsequent quantification of both arterial neointima
formation and
reendothelialization.25 We
have adapted this model of electric arterial injury to allow the
precise study of arterial reendothelialization and to determine which
ER subtype was involved in this process. Our results indicate that the
beneficial effect of estrogen on reendothelialization is mediated by
ER in female mice.
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Methods |
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TopAbstractIntroductionMethodsResultsDiscussionReferences |
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Animals
All procedures involving experimental animals were performed in accordance with the recommendations of the French Accreditation of Laboratory Animal Care. They were housed in stainless steel cages in groups of 5, kept in a temperature-controlled facility on a 12-hour light-dark cycle, and fed normal laboratory mouse chow diet.
Targeted disruption (knockout) of mouse ER and ERß
genes was generated by homologous recombination, resulting in ER-
and ERß-null mice.25A Six
backcrosses with C57Bl/6 mice were performed.
For all surgical procedures, mice were anesthetized by injection of 150 mg/kg ketamine IP and allowed to recover on a 37°C heat pack. Mice were ovariectomized at 4 weeks of age and given either 60-day time-release E2 pellets (0.1 mg E2, Innovative Research of America, ie, releasing 80 µg · kg-1 · d-1) or placebo-containing pellets implanted subcutaneously into the backs of the animals with a sterile trochar. Electric carotid artery injury was performed 2 weeks later, ie, in 6-week-old mice. Three days after injury, the animals were killed and the vessels harvested.
Electric Injury Model
We adapted the electric injury model described by
Carmeliet et al25 on the
femoral artery to the common carotid artery, the latter being easier to
dissect. Surgery was carried out with a dissection microscope (Nikon
SMZ-2B) in 6-week-old female mice weighing 20 g on average.
Because the proximal part of the carotid artery is intrathoracic, the
injury could not be applied to the whole common carotid artery. The
left common carotid artery was exposed via an anterior incision of the
neck. The electric injury (in fact primarily thermal) was applied to
the distal part of the common carotid artery. The carotid artery was
injured by electric current with a bipolar microregulator. To
standardize the temperature increase in the vessel wall, we used
forceps with large tips (1 mm) instead of microsurgical forceps (200
µm) and a bipolar microregulator Force FX (Valleylab). The
"precise" mode of this apparatus allowed delivery of electric
energy within a narrow range of resistance, because the generator
microprocessor disrupted the electric current when the resistance
increased as a consequence of temperature increase. This allowed the
increase in tissue temperature to be controlled and avoided the risk of
desiccation and coagulation of the arterial wall. The optimal
conditions were determined as follows: electric current of 2 W applied
for 2 seconds to each millimeter of carotid artery over a total length
of 4 mm with the help of a size marker placed parallel to the long axis
of the carotid. Despite optimization of the technique, coagulation and
thrombosis of the carotid artery occurred in 
10% of the cases,
which were then excluded from the study.
One to 7 days later, the endothelial regeneration process
was evaluated by staining the denuded areas with Evans blue dye as
previously described.23
Briefly, 50 µL of solution containing 5% Evans blue diluted in
saline was injected into the tail vein with a 30-gauge needle 10
minutes before euthanization, followed by fixation with a perfusion of
4% phosphate-buffered formalin (pH 7.0) for 5 minutes. Blood, saline,
and fixative were removed through an incision in the right atrium. The
left common carotid artery was dissected with an adjacent portion of
the aortic arch and carotid bifurcation. The artery was then opened
longitudinally and placed between slides with Fluoprep. After
transparency scanning and numeration, the total and stained carotid
artery areas were planimetered with an image analyzer
(VISIOL@b2000). The ratio between the area stained in blue and
the total carotid artery area was calculated. The surface of the area
that remained deendothelialized was indexed to the total carotid artery
area to take into account the changes in vessel area due to both the
elasticity of the carotid artery and the flattening of the vessel
between slides. The coefficients of variation of the endothelial
regrowth were 2.6%, 4.5%, and 13.5% at days 1, 3, and 5,
respectively
(Figure 1C
).
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Histology and Scanning Electron
Microscopy
Arteries were also embedded in paraffin, and sections
perpendicular to the long axis of the carotid were cut from the
proximal, middle, and distal thirds of the injured carotid artery.
Sections were subjected to standard hematoxylin and eosin
staining.
To verify that the area stained with Evans blue corresponded to the deendothelialized area and that the unstained area corresponded to the reendothelialized area, 3 animals were killed 3 days after the electric injury procedure. Perfusion fixation was carried out as described above with phosphate-buffered 4% paraformaldehyde in vivo. The vessels were cut open longitudinally, pinned flat on a silicone-coated dish, and photographed. The vessels were then further fixed with 2% glutaraldehyde for 24 hours, rinsed with PBS, incubated with 2% osmium tetroxide, and then dehydrated through a series of ethanol dilutions. The tissue was dried to critical point and mounted on scanning electron microscopy stubs with colloidal silver paste. After having been sputter-coated with gold/palladium, the specimens were examined with a scanning electron microscope.
Serum Hormone Concentrations
Radioimmunoassay kits for E2
were used according to the manufacturer’s instructions (Sorin
Biomedica). Hormone levels could not be measured in the same mice that
were injected with Evans blue because it interfered with the assay.
Thus, additional groups of mice were ovariectomized at 4 weeks of age,
given either E2- or placebo-containing pellets
as described above, and killed 17 days later. Hormone levels were
assayed for each individual mouse in a similar series of assays. The
intra-assay coefficient of variability was 4.5%. The assay
sensitivity, defined as 15% displacement of labeled tracer, was 0.5 pg
E2.
Statistics
Results are expressed as mean±SEM. To test the
respective roles of E2 treatment and of genotype
on reendothelialization, a 2-factor ANOVA was performed (comparison of
the 6 groups) in ER and in ERß mice. When an interaction was
observed between the 2 factors, the effect of E2
treatment was studied in each genotype with a
t test. A value of
P<0.05 was considered
statistically significant.
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Results |
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TopAbstractIntroductionMethodsResultsDiscussionReferences |
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Generation of the ER
and ERß Mutant
MiceHeterozygous ER
- and ERß-null mice, originally
generated as C57Bl/6Jx129 hybrids, were crossbred into the C57Bl/6
background to generate the heterozygous
ER+/- and
ERß+/- lines, which constituted the
population that served as the parental genotypes for all animals in the
studies. The offspring of the parental
ER+/- strain were
ER+/+,
ER+/-, and
ER-/-, and those of the parental
ERß+/- strain were
ERß+/+,
ERß+/-, and
ERß-/- littermates, respectively,
which served as the subjects of our studies.
Description of the Model of Carotid Artery
Injury
We first determined the time course of endothelial
regeneration (days 1, 3, 5, and 7 after injury) in 6-week-old female
C57Bl/6 mice. As shown in
Figure 1
, 50% of the endothelial regeneration was observed
on days 3 and 4, and complete reendothelialization was observed on day
7. We chose to study reendothelialization on day 3 to evaluate the
effect of E2. To further ensure that the area
stained with Evans blue dye corresponded to a deendothelialized area,
electron scanning microscopy studies were performed. These confirmed
that the area stained colocalized with an area of endothelial cell
denudation and a monolayer of aggregated platelets. A front of
migrating endothelial cells was observed at the edges of the lesion
(Figure 2A, 2B, and 2C).
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Paraffin sections (day 2) with standard staining
(hematoxylin/eosin and Masson’s trichrome) also demonstrated
the absence of endothelial cells (or rare pyknotic nuclei), the absence
of smooth muscle cells in the media (or only necrotic fragments), and
infiltration of the adventitia by inflammatory cells
(Figure 3a and 3b). Verhoeff staining for elastic lamina
revealed that only the most internal lamina was in some cases ruptured
(not shown). At day 15, ad integrum restoration of the intima
and of the media was observed
(Figure 3c and 3d). However, no neointimal hyperplasia was
observed after electric injury of the carotid, even on day 30
(Figure 3e and 3ff). Adventitia was still infiltrated by
inflammatory cells, which persisted at day 15
(Figure 3c and 3d) and at day 30
(Figure 3e and 3f).
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Effect of Estradiol on Endothelial
Regeneration
Ovariectomized mice with an implanted placebo pellet
showed nondetectable (<5 pg/mL ie,
20x10-12 mol/L) circulating levels of
E2, whereas those implanted with a pellet
releasing 0.1 mg E2 for 60 days (ie, 80
µg · kg-1 · d-1)
showed serum E2 concentrations averaging
0.5x10-9 mol/L, irrespective of genotype
(Tables 1 and 2). Ovariectomized, placebo-treated mice
showed atrophied uteri (<20 mg); E2-treated
mice showed a significant increase in uterine weight, except
ER-/- mice, which had atrophied uteri
even when treated with E2
(Tables 1 and 2).
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Endothelial regeneration was similar in all categories of
mice with an implanted placebo pellet, irrespective of genotype
(Tables 1 and 2,
Figure 4). In ER-deficient mice, an interaction
(P=0.004) between
E2 treatment and genotype on
reendothelialization was revealed by the 2-factor ANOVA. In these
animals, E2 treatment significantly accelerated
reendothelialization in ER+/+ mice
(P<0.0001), tended to
accelerate the reendothelialization in
ER+/- mice
(P=0.14), but had no effect in
ER-/- mice
(P=0.95)
(Figure 4 and
Table 1). In contrast, in ERß-deficient animals,
E2 treatment significantly accelerated the
reendothelialization process in ERß+/+,
ERß+/-, and
ERß-/- mice in a similar fashion
(P<0.0001), without influence
of genotype (P=0.27, NS).
Although the acceleration of the reendothelialization tended to be
greater in E2-treated
ERß-/- mice treated with
E2 than in ERß+/+
mice, the difference was not statistically significant, because no
interaction between E2 treatment and the
genotype was observed (P=0.73,
NS)
(Figure 4 and
Table 2).
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Discussion |
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TopAbstractIntroductionMethodsResultsDiscussionReferences |
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Endothelium is both a key actor in the pathophysiology of atherosclerosis and an important target of E2 in the vessel wall. Indeed, the modulation of several properties of endothelium in response to E2 could account for the vasculoprotective effect of the hormone. One major function of endothelium is the release of the vasorelaxant and antiaggregant messenger nitric oxide, which is enhanced in oophorectomized females treated with estrogens26 27 28 either directly by upregulation of the endothelial NO synthase gene expression29 or indirectly by a receptor-mediated antioxidant effect.30 E2 was also reported to promote reendothelialization in castrated female rats,16 but to the best of our knowledge, this effect has not been reported in mice. Moreover, the availability of genetically modified mice with ER
or ERß gene
disruption offers the possibility of studying the role of each of these
ER subtypes in the effect of E2 on endothelial
regeneration.
Preliminary experiments led us to modify the model of
electric injury initially described by Carmeliet et
al25 to study the
reendothelialization process. For technical reasons, we elected to
perform the electric injury in the carotid instead of the femoral
artery. As reported for the femoral
site,25 we found that the
reendothelialization process started immediately after injury and
proceeded very actively during the first week after injury. The 4 mm of
injured carotid artery was completely reendothelialized within 7 days,
ie, 300 µm/d for each edge. Taking into account that the average
length of an endothelial cell is 50 µm (see previous
reports31 32 and
Figure 2) and that the doubling time of this cell population
is in the range of 24 hours, proliferation would contribute to 50
µm/d from each edge over the 7-day period. It would appear from these
calculations that reendothelialization relies essentially on local cell
migration or circulating endothelial progenitor cells, which have been
shown to be incorporated into foci of
neovascularization.33 34
We observed that E2 accelerated
endothelial regrowth in wild-type mice by 25%, a value close to what
had been measured in the rat
species.16 We then sought to
determine which ER gene promoted the reendothelialization process.
ER has long been considered to be the unique target of
E2, because it has been characterized in
reproductive tissues, in bone, and in vessels, particularly in the
endothelial
cells.35 36 37
Iafrati et al24 reported
that the prevention of medial enlargement (ie, smooth muscle cell
proliferation) by E2 is preserved in
ER-deficient mice in a model of endovascular carotid injury. Very
recently, the same group reported that E2 also
inhibits medial enlargement in the injured carotid of ERß-deficient
mice.38 It should be
underlined that in these studies, vascular smooth muscle cell
proliferation was evaluated, whereas in our studies, the effect of
E2 on endothelial regeneration was appreciated.
Altogether, these series of data led to the intriguing possibility that
reendothelialization could be mediated specifically by the ER gene,
whereas arterial smooth muscle cell proliferation could be mediated by
a third, as yet unidentified ER gene. Further work should clarify this
situation, but taking into account the crucial role of endothelium in
the maintenance of vascular integrity, ER should be considered a
prime target of pharmacological studies in the area of cardiovascular
diseases.
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Acknowledgments |
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The work at INSERM U397 was supported in part by INSERM, the Ministère de la Recherche et de la Technologie, the Fondation de France, the Fondation de l’Avenir, and the Conseil Régional Midi-Pyrénées. The work at the Institut de Génétique et de Biologie Moléculaire et Cellulaire was supported by a grant from the CNRS, INSERM, the Collège de France, the Hôpital Universitaire de Strasbourg, the Association pour la Recherche Médicale, the Fondation pour la Recherche Médicale, and by GIP/HMR. Dr Dupont was supported by a fellowship from Hoechst Marion Roussel. We thank J.-L. Fontanilles, G.B.M., Hôpital Rangueil, for his valuable technical assistance.
Received May 22, 2000; revision received July 11, 2000; accepted August 1, 2000.
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J.-B. Michel, O. Thaunat, X. Houard, O. Meilhac, G. Caligiuri, and A. Nicoletti Topological Determinants and Consequences of Adventitial Responses to Arterial Wall Injury Arterioscler Thromb Vasc Biol, June 1, 2007; 27(6): 1259 - 1268. [Abstract] [Full Text] [PDF]
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D. Xing, W. Feng, A. P. Miller, N. M. Weathington, Y.-F. Chen, L. Novak, J. E. Blalock, and S. Oparil Estrogen modulates TNF-{alpha}-induced inflammatory responses in rat aortic smooth muscle cells through estrogen receptor-beta activation Am J Physiol Heart Circ Physiol, June 1, 2007; 292(6): H2607 - H2612. [Abstract] [Full Text] [PDF]
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B. Dawn and R. Bolli Increasing Evidence That Estrogen Is an Important Modulator of Bone Marrow-Mediated Cardiac Repair After Acute Infarction Circulation, November 21, 2006; 114(21): 2203 - 2205. [Full Text] [PDF]
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H. Hamada, M. K. Kim, A. Iwakura, M. Ii, T. Thorne, G. Qin, J. Asai, Y. Tsutsumi, H. Sekiguchi, M. Silver, et al. Estrogen Receptors {alpha} and {beta} Mediate Contribution of Bone Marrow-Derived Endothelial Progenitor Cells to Functional Recovery After Myocardial Infarction Circulation, November 21, 2006; 114(21): 2261 - 2270. [Abstract] [Full Text] [PDF]
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V. Fontaine, C. Filipe, N. Werner, P. Gourdy, A. Billon, B. Garmy-Susini, L. Brouchet, F. Bayard, H. Prats, T. Doetschman, et al. Essential Role of Bone Marrow Fibroblast Growth Factor-2 in the Effect of Estradiol on Reendothelialization and Endothelial Progenitor Cell Mobilization Am. J. Pathol., November 1, 2006; 169(5): 1855 - 1862. [Abstract] [Full Text] [PDF]
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C. Bolego, E. Vegeto, C. Pinna, A. Maggi, and A. Cignarella Selective Agonists of Estrogen Receptor Isoforms: New Perspectives for Cardiovascular Disease Arterioscler Thromb Vasc Biol, October 1, 2006; 26(10): 2192 - 2199. [Abstract] [Full Text] [PDF]
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T. Simoncini, C. Scorticati, P. Mannella, A. Fadiel, M. S. Giretti, X.-D. Fu, C. Baldacci, S. Garibaldi, A. Caruso, L. Fornari, et al. Estrogen Receptor {alpha} Interacts with G{alpha}13 to Drive Actin Remodeling and Endothelial Cell Migration via the RhoA/Rho Kinase/Moesin Pathway Mol. Endocrinol., August 1, 2006; 20(8): 1756 - 1771. [Abstract] [Full Text] [PDF]
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D. Seetharam, C. Mineo, A. K. Gormley, L. L. Gibson, W. Vongpatanasin, K. L. Chambliss, L. D. Hahner, M. L. Cummings, R. L. Kitchens, Y. L. Marcel, et al. High-Density Lipoprotein Promotes Endothelial Cell Migration and Reendothelialization via Scavenger Receptor-B Type I Circ. Res., January 6, 2006; 98(1): 63 - 72. [Abstract] [Full Text] [PDF]
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R. G. Mishra, F. Z. Stanczyk, K. A. Burry, S. Oparil, B. S. Katzenellenbogen, M. L. Nealen, J. A. Katzenellenbogen, and R. K. Hermsmeyer Metabolite ligands of estrogen receptor-{beta} reduce primate coronary hyperreactivity Am J Physiol Heart Circ Physiol, January 1, 2006; 290(1): H295 - H303. [Abstract] [Full Text] [PDF]
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H. Tan, X. Jiang, F. Yang, Z. Li, D. Liao, J. Trial, M. J. Magera, W. Durante, X. Yang, and H. Wang Hyperhomocysteinemia inhibits post-injury reendothelialization in mice Cardiovasc Res, January 1, 2006; 69(1): 253 - 262. [Abstract] [Full Text] [PDF]
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K. Wassmann, S. Wassmann, and G. Nickenig Progesterone Antagonizes the Vasoprotective Effect of Estrogen on Antioxidant Enzyme Expression and Function Circ. Res., November 11, 2005; 97(10): 1046 - 1054. [Abstract] [Full Text] [PDF]
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K. Kublickiene, E. Svedas, B.-M. Landgren, M. Crisby, N. Nahar, H. Nisell, and L. Poston Small Artery Endothelial Dysfunction in Postmenopausal Women: In Vitro Function, Morphology, and Modification by Estrogen and Selective Estrogen Receptor Modulators J. Clin. Endocrinol. Metab., November 1, 2005; 90(11): 6113 - 6122. [Abstract] [Full Text] [PDF]
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W. Koch, P. Hoppmann, A. Pfeufer, J. C. Mueller, A. Schomig, and A. Kastrati No Replication of Association Between Estrogen Receptor {alpha} Gene Polymorphisms and Susceptibility to Myocardial Infarction in a Large Sample of Patients of European Descent Circulation, October 4, 2005; 112(14): 2138 - 2142. [Abstract] [Full Text] [PDF]
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M. Zaitseva, D. S. Yue, J. A. Katzenellenbogen, P. A. W. Rogers, and C. E. Gargett Estrogen Receptor-{alpha} Agonists Promote Angiogenesis in Human Myometrial Microvascular Endothelial Cells Reproductive Sciences, December 1, 2004; 11(8): 529 - 535. [Abstract] [PDF]
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Y. Nakamura, K. Igarashi, T. Suzuki, J. Kanno, T. Inoue, C. Tazawa, M. Saruta, T. Ando, N. Moriyama, T. Furukawa, et al. E4F1, a Novel Estrogen-Responsive Gene in Possible Atheroprotection, Revealed by Microarray Analysis Am. J. Pathol., December 1, 2004; 165(6): 2019 - 2031. [Abstract] [Full Text] [PDF]
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A. Abizaid, M. Albertal, M. A. Costa, A. S. Abizaid, R. Staico, F. Feres, L. A. Mattos, A. G. M. R. Sousa, J. Moses, N. Kipshidize, et al. First human experience with the 17-beta-estradiol-eluting stent: The estrogen and stents to eliminate restenosis (EASTER) trial J. Am. Coll. Cardiol., March 17, 2004; 43(6): 1118 - 1121. [Abstract] [Full Text] [PDF]
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A. Iwakura, C. Luedemann, S. Shastry, A. Hanley, M. Kearney, R. Aikawa, J. M. Isner, T. Asahara, and D. W. Losordo Estrogen-Mediated, Endothelial Nitric Oxide Synthase-Dependent Mobilization of Bone Marrow-Derived Endothelial Progenitor Cells Contributes to Reendothelialization After Arterial Injury Circulation, December 23, 2003; 108(25): 3115 - 3121. [Abstract] [Full Text] [PDF]
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M. Kimura, K. Sudhir, M. Jones, E. Simpson, A.-M. Jefferis, and J. P.F. Chin-Dusting Impaired Acetylcholine-Induced Release of Nitric Oxide in the Aorta of Male Aromatase-Knockout Mice: Regulation of Nitric Oxide Production by Endogenous Sex Hormones in Males Circ. Res., December 12, 2003; 93(12): 1267 - 1271. [Abstract] [Full Text] [PDF]
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A. M. Shearman, L. A. Cupples, S. Demissie, I. Peter, C. H. Schmid, R. H. Karas, M. E. Mendelsohn, D. E. Housman, and D. Levy Association Between Estrogen Receptor {alpha} Gene Variation and Cardiovascular Disease JAMA, November 5, 2003; 290(17): 2263 - 2270. [Abstract] [Full Text] [PDF]
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P. Geraldes, M. G. Sirois, and J.-F. Tanguay Specific Contribution of Estrogen Receptors on Mitogen-Activated Protein Kinase Pathways and Vascular Cell Activation Circ. Res., September 5, 2003; 93(5): 399 - 405. [Abstract] [Full Text] [PDF]
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K. Strehlow, N. Werner, J. Berweiler, A. Link, U. Dirnagl, J. Priller, K. Laufs, L. Ghaeni, M. Milosevic, M. Bohm, et al. Estrogen Increases Bone Marrow-Derived Endothelial Progenitor Cell Production and Diminishes Neointima Formation Circulation, June 24, 2003; 107(24): 3059 - 3065. [Abstract] [Full Text] [PDF]
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G. A. Figtree, D. McDonald, H. Watkins, and K. M. Channon Truncated Estrogen Receptor {alpha} 46-kDa Isoform in Human Endothelial Cells: Relationship to Acute Activation of Nitric Oxide Synthase Circulation, January 7, 2003; 107(1): 120 - 126. [Abstract] [Full Text] [PDF]
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J.-F. Arnal and F. Bayard Alteration in Endothelial Estrogen Receptor Expression: A Potential Key of Vasculoprotection by Estrogens? Circ. Res., November 1, 2002; 91(9): 759 - 760. [Full Text] [PDF]
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C. E. Gargett, M. Zaitseva, K. Bucak, S. Chu, P. J. Fuller, and P. A. W. Rogers 17{beta}-Estradiol Up-Regulates Vascular Endothelial Growth Factor Receptor-2 Expression in Human Myometrial Microvascular Endothelial Cells: Role of Estrogen Receptor-{alpha} and -{beta} J. Clin. Endocrinol. Metab., September 1, 2002; 87(9): 4341 - 4349. [Abstract] [Full Text] [PDF]
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Y. Yuan, L. Liao, D. A. Tulis, and J. Xu Steroid Receptor Coactivator-3 Is Required for Inhibition of Neointima Formation by Estrogen Circulation, June 4, 2002; 105(22): 2653 - 2659. [Abstract] [Full Text] [PDF]
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G. Pare, A. Krust, R. H. Karas, S. Dupont, M. Aronovitz, P. Chambon, and M. E. Mendelsohn Estrogen Receptor-{alpha} Mediates the Protective Effects of Estrogen Against Vascular Injury Circ. Res., May 31, 2002; 90(10): 1087 - 1092. [Abstract] [Full Text] [PDF]
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B. Darblade, C. Pendaries, A. Krust, S. Dupont, M.-J. Fouque, J. Rami, P. Chambon, F. Bayard, and J.-F. Arnal Estradiol Alters Nitric Oxide Production in the Mouse Aorta Through the {alpha}-, but not {beta}-, Estrogen Receptor Circ. Res., March 8, 2002; 90(4): 413 - 419. [Abstract] [Full Text] [PDF]
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C. Pendaries, B. Darblade, P. Rochaix, A. Krust, P. Chambon, K. S. Korach, F. Bayard, and J.-F. Arnal The AF-1 activation-function of ERalpha may be dispensable to mediate the effect of estradiol on endothelial NO production in mice PNAS, February 19, 2002; 99(4): 2205 - 2210. [Abstract] [Full Text] [PDF]
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B. Darblade, C. Pendaries, A. Krust, S. Dupont, M.-J. Fouque, J. Rami, P. Chambon, F. Bayard, and J.-F. Arnal Estradiol Alters Nitric Oxide Production in the Mouse Aorta Through the {alpha}-, but not {beta}-, Estrogen Receptor Circ. Res., March 8, 2002; 90(4): 413 - 419. [Abstract] [Full Text] [PDF]
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