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(Circulation. 1995;92:2975-2983.)
© 1995 American Heart Association, Inc.
Articles |
Progesterone Receptor Expression in Human Saphenous Veins
From INSERM U 135, Hormones et Reproduction Faculté de Médecine Paris Sud, Le Kremlin-Bicêtre (M.P.-A., K.C.-S., E.M.) and Innothera (M.F.), Arcueil, France.
Correspondence to Dr Martine Perrot-Applanat, INSERM U.55, Centre de Recherche Paris Saint Antoine, 184 rue du Frubourg Saint Antoine, 75571 Paris, Cedex 12, France.
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Abstract |
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 Top Abstract IntroductionMethods Results Discussion References |
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Background Clinical and epidemiological observations regarding varicose veins, such as their predominance in women and the occurrence of venous stasis during sex-hormone therapy, the luteal phase of the menstrual cycle, and pregnancy, suggest a sex hormone–dependency of this venous pathology. In the present study, analysis of steroid receptors was used to determine if these effects were due to a direct hormonal action on the saphenous vein.
Methods and Results Biopsy samples were obtained from patients undergoing stripping removal of varicose saphenous veins. Patients were men (n=5) and premenopausal (n=15) or postmenopausal (n=10) women. Progesterone receptors (PR) and estrogen receptors (ER) were determined by both enzyme immunoassay (EIA) and immunocytochemistry by use of monoclonal antibodies. Ninety percent of the biopsy samples showed PR positivity by EIA (range, 5 to 53 fmol/mg cytosol protein). When present, PR staining was observed in the cell nuclei of the tunica media and the subendothelial layer (neointima). No significant variation was observed in the PR content of different regions within the same saphenous vein. In contrast, no ER or extremely low levels of ER (<5 fmol/mg cytosol protein) were detected by EIA in 25 of 30 varicose biopsy samples. Reverse transcription–polymerase chain reaction (RT-PCR) was used to analyze PR and ER mRNAs in biopsy samples that were PR positive/ER negative. With primers to the hormone-binding region encoded by PR mRNA, a RT-PCR product of the expected size was detected and its identity confirmed by Southern blot by use of a PR cDNA probe. In contrast, no RT-PCR product could be detected by use of primers to the DNA-binding domain, the hinge region, and the ligand-binding domain encoded by ER mRNA.
Conclusions These results indicate that human saphenous veins from both sexes express PR, as previously described for arterial blood vessels. This observation suggests that progesterone acts directly on these veins via a classic receptor-mediated pathway.
Key Words: veins • biopsy • polymerase chain reaction • immunoassays • estrogen receptors
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Introduction |
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TopAbstractIntroductionMethods Results Discussion References |
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The prevalence and incidence of varicose veins appears to vary across populations, reaching higher levels in industrialized Western societies and causing various health problems, including deep-vein thrombosis and ulcers. Various hypotheses concerning the causes of this venous pathology have been proposed, including heredity, prolonged sitting and standing, dietary factors, obesity, and atherosclerosis.1 2 Obesity, low levels of physical activity, high systolic blood pressure, and cigarette smoking (for men) were linked to the recurrence of varicose veins among the 5209 male and female subjects of the Framingham Study.3 In addition, the predominance of varicose veins in women1 3 and the occurrence of venous stasis during luteal phase, pregnancy, or sex-hormone therapy have suggested a sex-hormone dependency of this venous pathology. However, it is not known whether estradiol and/or progesterone act directly on the venous system or if they have an indirect effect occurring, for example, through modifications of blood coagulability. The detection of steroid-hormone receptors in the vessel walls would provide strong evidence for a direct effect of these hormones.
In target tissues, sex hormones exert their effects through receptor proteins located in the nuclear compartment of the cells.4 5 In most target tissues, ie, the uterus and the mammary gland, the synthesis of ER and PR is stimulated by estrogens, through the available estrogen receptors.4 Recently, the presence of PR and ER in the arterial blood vessels of the reproductive tract has been reported,6 7 suggesting that sex steroids exert an effect on the modulation of uterine blood flow. The concentration of arterial receptors appears to be hormonally modulated.6 ER have also been detected in coronary arteries and vascular smooth muscle cells cultured from surgical specimens of mammary arteries.8 9 Moreover, the expression of ER in coronary arteries has been related to the absence of atherosclerosis in premenopausal women.8
To evaluate whether sex steroids may have a direct effect on saphenous veins, we used immunocytochemistry and EIA to investigate the presence of ER and PR in biopsy samples obtained from female and male patients during "stripping" surgery. The results of these studies were confirmed at the mRNA level by use of RT-PCR.
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Methods |
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TopAbstractIntroductionMethods Results Discussion References |
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Tissue Collection
Thirty biopsy samples were obtained from patients undergoing surgical removal of varicose saphenous veins. The fragments were trimmed of adhering fat and connective tissue, quickly frozen in liquid nitrogen, and stored until analysis. Biochemical analysis, immunocytochemical staining for ER and PR, and detection of mRNA coding for these receptors were performed on separate fragments (1 cm long) from each biopsy sample. Among the 30 biopsy samples examined, 15 were collected from premenopausal women (26 to 48 years old), 10 from postmenopausal women (48 to 78 years old), and 5 from men (45 to 54 years old). Two additional biopsy samples of normal saphenous veins were obtained from 2 patients (one male and one female) undergoing coronary artery bypass surgery. Positive and negative controls, provided by uterine (or breast) carcinoma and spleen tissue, respectively, were included in the study.
PR and ER EIA
PR and ER were measured in the cytosols from
human saphenous
veins by EIA with commercial anti-PR or anti-ER antibodies coated on
beads (Abbott Laboratories). Receptor values were reported as the
number of femtomoles of ER or PR bound per milligram of cytosol
protein. Biopsy samples with receptor content <5 fmol/mg cytosol
protein were considered to be receptor-negative.
Immunocytochemistry
Two monoclonal antibodies to human ER
(ERICA kit, Abbott
Laboratories) and PR (Li 417)10 (available from Transbio)
were used for this study. These antibodies have been extensively
characterized and their specificity studied by immunochemical and
immunocytochemical
methods.10 11 12 13 14
The epitope recognized by
Li 417 is located in the N-terminal region (within aa 208 to 296) of
PR.10 The epitope recognized by the ER antibody is located
in the hormone-binding domain.13 Immunocytochemical
detection of ER and PR was performed as previously
described.7 11 12 13 14
Sections (5 µm) of frozen tissue were
cut at -25°C and fixed in picric acid formaldehyde for 20 minutes at
4°C. Sections were incubated for 18 hours at 4°C with primary
antibodies and for 1 hour at room temperature with secondary
antibodies. PR was immunostained with anti-human PR
antibody Li 417 (6 µg/mL), biotinylated goat anti-mouse antibody
(Amersham; diluted 1:100), and a streptavidin-biotin-peroxidase
complex (Amersham; diluted 1:100). ER was immunostained
with the Abbott kit, with slight modifications as follows: overnight
incubation at 4°C with monoclonal ER antibody, followed by incubation
for 1 hour at room temperature with goat anti-rat IgG and rat
monoclonal peroxidase-antiperoxidase complexes. Peroxidase activity
was revealed by the diaminobenzidine reaction (0.7 mg/mL in 0.025%
hydrogen peroxide for 10 minutes) and observed by means of light
microscopy. The staining intensity of individual cells was
characterized as absent (–), weak (+), moderate (++), or
intense
(+++). Duplicate sections were lightly counterstained with
hematoxylin
to facilitate the identification of cellular elements.
Controls
A section of each tissue adjacent to the
immunostained section was treated by the immunoperoxidase
method, but the primary antibodies were replaced by unrelated mouse- or
rat-receptor antibodies.6
Localization of ER and PR in Saphenous Veins
The various
venous cell types were identified with use of
specific antibodies. Endothelial cells of the intima
were labeled with the monoclonal antibody BNH9 (Immunotech). This
antibody, which is reactive with vascular and lymphatic
endothelial cells, recognizes a blood
group–related antigen carried by H and Y determinants.
Subendothelial cells of the intima were labeled
with a monoclonal anti-
-smooth muscle actin antibody (Sigma Chem
Co). Myocytes (smooth muscle cells) in the media were labeled with a
rabbit anti-desmin antibody (Eurodiagnostic) and
the anti--smooth muscle actin antibody. Adventitial fibroblasts
were labeled with anti-vimentin antibody (ICN Immunobiologicals).
Immunocytochemical detection of these specific markers was performed as
previously described.7 These antibodies were applied on
frozen sections adjacent to those processed for steroid receptors.
RNA Preparation
Total RNA was extracted from human saphenous
vein tissue (0.8 to
2.2 g) according to the protocol outlined in the RNA ß kit (Bioprobe
Systems). SvRNA was quantified by absorbance at 260 nm,
analyzed by electrophoresis in 1% agarose to establish RNA
integrity, and stored at -70°C until use. RNA from human uterine
tissue and from human spleen were used as positive and negative
controls, respectively.
cDNA Synthesis
cDNA was synthesized from total RNA (4
µg). Reactions were
carried out with 40 U RNAse inhibitor and 80 U reverse
transcriptase with the cDNA synthesis System Plus (Amersham) in 40 µL
final volume, according to the manufacturer's instructions.
Incubations were carried out for 1 hour at 42°C.
Oligonucleotide Primers and Probes
The following primer
sequences corresponding to human ER
cDNA15 were used in the RT-PCR to detect ER mRNA: (a)
amplification of cDNA corresponding to exons 1 through
5,16 primers 1a and 1b; 1a, sense
5'-CCCCACGGCCAGCAGGTGCCCTACT-3' (aa 122 through 129);
1b, anti-sense 5'-GAGCGCCAGACGAGACCAATCATCA-3'
(aa 387 through 395); (b) amplification of exons 4 through 8, primers
2a and 2b; 2a, sense 5'-TGATGGGGAGGGCAGGGGTGAAGTG-3' (aa 272
through
279); 2b, anti-sense 5'
-TAGGCGGTGGGCGTCCAGCATCTCC-3' (aa
541 through 549). The following oligonucleotides
corresponding to human PR cDNA17 were also used: (c)
amplification of cDNA corresponding to exons 4 through
8,18 primers 3a and 3b; 3a, sense
5'-GTGGGCGTTCCAAATGAAAGCCAAG-3' (aa 660
through 667); 3b, anti-sense 5'-AATTCAACACTCAGTGCCCGGGACT-3' (aa
897 through 905); (d) amplification of exons 1 through 4, primers 4a
and 4b; 4a, sense
5'-ACCCGCCAGTGCCTCAGTCTCGTCT-3'
(aa 443 through 450); 4b, anti-sense
5'-GGCTTTCATTTGGAACGCCCACTGG-3' (aa 659
through 666). Oligonucleotides were synthesized by use
of an Applied Biosystems DNA synthesizer. Hybridization of PCR
products was performed with PR cDNA17 and ER cDNA.
Human ER cDNA (gift from H. Loosfelt) has been cloned from a cDNA
library of T47D cells (F. Fridlanski and H. Loosfelt). Its sequence is
identical to that of the native (wild-type)
receptor.15 Probes were labeled by use of
[32P]-dCTP and Kleenow enzyme with the random primed
DNA labeling kit (Boehringer Mannheim Corp) to a specific
activity of 5x108 cpm/µg cDNA.
PCR and Southern Blot Analysis
One fourth of the cDNA
reaction mixture was combined with 10
µL of 10x PCR buffer (Perkin Elmer/Cetus), 0.2 mmol/L dNTP
(Boehringer Mannheim Corp), 150 ng of each primer, and 2.5 U of
Taq polymerase (Perkin Elmer/Cetus) in a final volume of 100
µL. Primers 1a and 1b (exons 1 through 5) or primers 2a and 2b (exons
4 through 8) were used to amplify ER cDNA. Primers 3a and 3b (exons 4
through 8) were used to amplify PR cDNA. Each cycle of amplification
consisted of 1 minute of denaturation (94°C) followed by 1 minute of
annealing (55°C) and 1 minute of extension (72°C). Each PCR
consisted of 30 cycles.
A 25-µL aliquot of the PCR reaction mixture
was size-fractionated
by electrophoresis on 1% agarose gels. An aliquot of a DNA ladder
(
x174 DNA-Hae III digest, Biolabs) was included as a
size marker. After electrophoresis, gels were denatured for 30 minutes
in 0.5 mol/L NaOH and 1.5 mol/L NaCl and subsequently neutralized for
30 minutes in a 1.5 mol/L NaCl, 0.5 mol/L Tris buffer. After capillary
transfer on nitrocellulose filter (Schleicher et Schuell) overnight,
the filter was baked (2 hours at 80°C). Hybridization of PCR
products with ER or PR cDNA probe was performed at 60°C for 14
hours, and the filter was washed in 2-0.1xSSC, 0.5% SDS 60°C before
exposure to a Kodak direct-exposure film.
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Results |
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TopAbstractIntroductionMethods Results Discussion References |
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To study sex-steroid receptors in human varicose veins, we used EIAs of tissue extracts and the immunoperoxidase method on frozen sections.
EIA of ER and PR
Tables 1
and 2
summarize the results
of PR and ER assays in 30 human varicose veins. Ninety percent of the
biopsy samples contained PR (mean, 18 fmol/mg cytosol protein; range, 5
to 53 fmol/mg cytosol protein). In contrast, no ER or extremely low
levels of ER (<5 fmol/mg cytosol protein) were detected by EIA in 25
of 30 biopsy samples. Only 5 biopsy samples contained low levels of ER
(6 to 9 fmol/mg protein). Most varicose biopsy samples (70%) were
considered as ER–/PR+, whereas only 20% of samples were
ER+/PR+ and
10% were ER–/PR–.
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The presence of PR and absence or low
level of ER was further
analyzed in relation to sex, menopausal status, and
localization within the biopsy sample. The presence of PR was observed
in 92% of women versus 80% of men and in 100% of the premenopausal
patients versus 80% of the postmenopausal women (Table 2
). In
four
patients (three women, one man), the presence of receptors was
analyzed in three different samples obtained from the upper
(proximal), median, or lower (distal) regions of the removed saphenous
vein. Tissues analyzed in this experiment were all ER–/PR+, a
situation encountered in most samples analyzed. No significant
variation was observed in ER or PR contents from different regions
within the same saphenous vein. A similar observation was made for two
saphenous veins of one patient, with no correlation of the level of PR
(or ER) detected with the extent of varicosity (Table 3).
Similarly, no significant variation was observed in
receptor content from varicose fragments or fragments free of
varicosity taken from the same patient (Table 4).
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PR and
ER were also analyzed in two normal saphenous veins,
obtained from patients undergoing cardiac surgery. One patient was
ER–/PR– and the other was ER–/PR+ (see Table
1).
Immunostaining for ER and PR
Eleven samples were examined
with use of both ER and PR
immunoperoxidase methods. These methods have been extensively validated
previously in several human tissues, including blood
vessels.6 7 11 12 13 14
Seven samples were ER– (<5 fmol/mg
protein) and PR+ (5 to 53 fmol/mg protein), two were ER–/PR–,
and two
were ER+/PR+, as determined by EIA (Table 1).
Specific
immunostaining for PR was present in five cases
(which were also positive in the EIA assay), whereas PR staining was
not detectable in the biopsy samples containing no PR or very low
concentrations of PR (below 11 fmol PR/mg cytosol protein; see cases
no. 1, 7, and 13 in Table 1). ER staining was detectable in
none of
these cases. The ER+/PR+ samples (cases no. 13 and 15), which gave
no
ER staining, contained low concentrations of ER (6 to 9 fmol/mg
protein). The EIA ER–/PR– samples (cases no. 11 and 14) gave no ER
or
PR staining. In contrast, two different positive controls, breast tumor
and endometrial samples known to express ER,7 11 were
used
in all staining runs and showed ER immunolabeling (Fig 1e). The
specificity of the
immunostaining was also demonstrated in vascular cells
by the absence of labeling in sections incubated with unrelated
monoclonal antibodies or after preadsorption of anti-PR antibodies with
purified receptor as previously described.11 12
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In all saphenous veins tested, PR immunostaining was
observed in cell nuclei of the tunica media and of the
subendothelial layer (neointima) (Fig 1a through 1d).
Fibroblasts from the tunica adventitia were stained
occasionally (Fig 1a). Smooth muscle cells of the nutritive
arteries
present in the adventitia of the vein were also stained for PR (Fig
1a). Cellular localization of PR in the wall of saphenous veins
was
further investigated by use of specific markers of
endothelial cells, smooth muscle cells, and
fibroblasts. Cells that stained for PR were confirmed to be myocytes of
the tunica media (Fig 2) and the
subendothelial layer. This preferential
distribution of PR in the smooth muscle cells was identical in all
biopsy samples tested. Although the number of cases was limited,
samples containing lower PR values (11 to 30 fmol/mg protein) showed a
lower number of stained smooth muscle cells than samples containing
higher PR values (30 to 50 fmol/mg protein). In contrast,
endothelial cells were devoid of PR in all
specimens.
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ER and PR mRNA in Varicose Saphenous Veins
The presence of
relatively low amounts of PR in varicose veins
suggested that PR mRNA (and ER mRNA) were also in low supply, and this
led to the choice of PCR for its study. We wanted to examine the
presence of PR mRNA in the PR+ biopsy samples, even when ER was
absent (70% of total PR+ samples). The specificity of the RT–PCR
has
been validated in preliminary experiments performed with utRNA
(see "Methods").
Total RNA was isolated from eight
ER–/PR+ biopsy samples of human
varicose veins (PR content, 5 to 27 fmol/mg cytosol protein; see cases
no. 23 through 30 in Table 1) and reverse-transcribed to cDNA.
After PCR amplification of PR cDNA (with use of primers 3a and 3b,
covering exons 4 to 8, which correspond to the hormone-binding
domain, aa 660 through 905, as described in "Methods") and
hybridization with a PR cDNA probe, a specific product was detected
in the samples (Figs 3 and 4A). This PCR
product had the expected size of 737 bp, similar to the PCR
product generated by amplification of cDNA from the uterus. A
specific PCR product of 669 bp was obtained in the same samples
with use of primers 4a and 4b, covering exons 1 to 4. This corresponds
to the N-terminal region, the DNA-binding segment, and part of the
hormone-binding domain (aa 443 to 666). The spRNA did not yield
such amplification products.
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The expression of ER mRNA was examined in
the same samples, after PCR
amplification of the cDNA with use of either primers 1a and 1b or 2a
and 2b (covering exons 1 to 5 and 4 to 8, respectively) (Figs 3
and 4B). No amplification product was detected in any of the
RNA
samples prepared from varicose veins or from the spleen, in contrast to
the 818 or 832 bp products generated when utRNAs were used.
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Discussion |
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TopAbstractIntroductionMethods Results Discussion References |
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The data presented here show that human saphenous veins from women undergoing stripping surgery express PR mRNA and PR. A meaningful quantification of these receptors, which are present in low concentrations, was performed with use of a highly reproducible and sensitive assay (EIA). The specificity of the monoclonal antibodies and the validity of immunostaining of PR in various progesterone target or nontarget cells have been extensively analyzed by our group in previous reports.6 7 10 11 12 13 14 Specificity of the immunostaining also was demonstrated in arterial6 7 and venous cells by the absence of labeling of sections incubated with receptor-unrelated monoclonal antibody and by the complete absence of staining after preabsorbtion of anti-PR antibodies with purified receptor. We report the presence of PR in the tunica media (smooth muscle cells) from saphenous veins, as previously observed in arterial smooth muscle cells.6 7 19 20 Nuclear localization of PR is similar to that observed in vascular and nonvascular cells (see review in Reference 5). This localization in smooth muscle cells was observed in all PR+ samples, irrespective of their receptor content. Samples containing higher PR values as determined by EIA showed the highest number of stained smooth muscle cells. The absence of PR in endothelial cells of all varicose veins examined, as determined by immunocytochemistry, is in agreement with previous observations on arteries: spiral arteries of the human endometrium8 9 (by use of immunocytochemistry), coronary arteries and aorta of the baboon20 (by use of autoradiography) and dog aorta21 (by use of comparative receptor determinations in stripped intima or intact aorta). The relatively low levels (5 to 53 fmol/mg protein) of receptor in the saphenous veins is consistent with the moderate abundance of PR in other nonreproductive tissues, including arteries from the cardiovascular system6 19 20 21 22 23 and meningiomas.14 The presence or absence of PR (as determined by EIA) correlated well (91%) with immunocytochemical staining for PR in 11 varicose samples. The only exception were 2 samples that had a low concentration of PR (<11 fmol/mg cytosol protein). It has been shown previously11 that breast tumors containing low amounts of receptor may often be negative in immunocytochemical studies: the few cells that contain PR may not be present on the tissue sections that are examined. Experiments that used RT-PCR and two pairs of primers covering different parts of the receptor molecule confirmed the expression of PR mRNA in varicose veins.
In contrast, ER and its mRNA were undetectable in 80% of varicose veins by use of the same methods as those used for demonstrating the presence of PR. Only 5 of 30 biopsy samples contained ER in very low levels (6 to 9 fmol/mg protein), close to the sensitivity of the EIA. Although the number of cases was limited, the absence or low concentration of ER was not limited to a specific region of the vein, as shown by analysis of different regions within the same vein or by the study of the two saphenous veins taken from the same patient. That little (or no) ER could be detected in varicose veins has been reported previously in 31 patients by Schmidt et al,24 who used a ligand-binding method. The sensitivity of our assay for ER (<5 fmol/mg protein), which is higher than the sensitivity of the binding techniques, may explain the quantitative difference (16% versus 3% of ER) between the two studies. Experiments with RT-PCR and several different controls confirmed that the mRNA encoding ER was undetectable (ie, under the limit of sensitivity of the assay) in the varicose samples shown to be ER–/PR+ at the protein level. First, ER was amplified from the uterus but not from the ER-negative spleen. Second, amplification of cDNA by use of utRNA resulted in products of the predicted size and in their hybridization with a specific probe. Finally, results were similar with use of two different pairs of primers to amplify ER (primers 1a and 1b corresponding to aa 122 to 395 and primers 2a and 2b corresponding to aa 272 to 549). These primers cover part of the N-terminal region, the DNA-binding domain, the hinge region, and the ligand-binding domain of the receptor. PCR performed in these conditions excluded the presence of exon deletion mutants, such as mutants deleted in exons 4, 5, or 7, which have been found in some PR +/ER– breast cancers, breast cancer cell lines, and meningiomas.25 26
The present study, performed on 30 patients, showed that 90% of the varicose biopsy samples contained PR (range, 5 to 53 fmol/mg protein). Of these, 70% contained PR in the absence of ER. These results raise several questions regarding the mechanisms of regulation of PR and the functional significance of the presence of these receptors. Transcriptional regulation of the PR gene involves induction by estrogens and downregulation by progestins in most target cells.27 28 29 In contrast, synthesis of PR does not seem to be estrogen-dependent in varicose veins, as demonstrated for meningiomas,30 sublines of T47D human breast cancer cells,31 32 and IK 90 human endometrial cancer cells.33
The absence (or low level) of ER in most varicose veins, as described
in the present study, could be specific to the venous system or
could be due to a loss of the receptor during the degenerative process.
Chronic venous insufficiency is known to be correlated with cell
hypertrophy, transformation of contractile smooth muscle
cells into proliferating metabolic cells, appearance of
fibrosis and necrosis, and alterations of
microcirculation.34 However, our preliminary observations
have shown no differences in ER and PR content of varicose fragments
versus fragments free of varicosity in the veins of the same patients.
Moreover, ER was absent (or present at a very low level) in normal
saphenous veins obtained from two patients undergoing coronary
artery bypass. In contrast, smooth muscle cells freshly cultured from
surgical specimens of human saphenous veins taken in patients
undergoing coronary artery bypass surgery35 have
previously been shown to contain low levels of ER.9
However, a comparison with the present study is difficult because
the number of patients, their age, and the healthy or pathological
status of the veins were not indicated in the latter study. Culture
conditions also may have altered the phenotype of smooth
muscle cells. Such changes in -adrenergic receptor subtypes
between thoracic aorta media and vena cava and cultured vascular smooth
muscle cells derived from the same sources have been
described.36 Finally, smooth muscle cells in tissue
culture also could have come from feeder arteries, at least in part.
Whether estrogen affects smooth muscle cell growth, as has been
reported for arterial smooth muscle
cells,37 38 39 40 remains to
be clarified.
The presence of PR in the media of saphenous veins suggests that progesterone may influence venous structure and/or function. Physiological venous changes have been described on the basis of hormonal status, such as venous stasis during luteal phase or pregnancy. Consequently, venous blood flow may be directly modulated by steroid hormones. Changes in uterine blood flow, known to occur in the cycling uterus or during pregnancy,41 were previously shown to be directly modulated by steroid hormones via their specific receptors (ER and PR) present in vascular smooth muscle cells.6 7 In particular, the presence of PR staining in smooth muscle cells of spiral arteries in early pregnancy7 correlates with large increases in endometrial and myometrial blood flow that occur to support fetal growth and homeostasis. The role of progesterone in the saphenous vein remains to be determined. We have reported preliminary data on the in vitro progesterone-induced decrease of noradrenaline and calcium-induced contractions from isolated human saphenous veins.42 Whether or not progesterone has also an effect on the structure of the vein via a regulation of cell proliferation remains unknown.
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Selected Abbreviations and Acronyms |
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Acknowledgments |
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This work was supported by the INOV foundation, the Institut National de la Santé et de la Recherche Medicale, the Centre National de la Recherche Scientifique, the UER Kremlin Bicêtre and the Fondation pour la Recherche Médicale Française. We thank Dr Michel Atger and Dr Hugues Loosfelt (INSERM U135, France) for their help in RT-PCR analysis. Catherine Blonde typed the manuscript.
Received November 9, 1994; revision received June 27, 1995; accepted July 3, 1995.
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References |
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TopAbstractIntroductionMethods Results Discussion References |
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