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ABSTRACT: OBJECTIVE: To investigate whether hypoxia or the female sex steroids exert direct effects on angiopoietin-1 (ANGPT1), ANGPT2, and vascular endothelial growth factor (VEGF) in human endometrial stromal cells (ESCs) to clarify the regulatory function of these local angiogenic factors in the endometrium. STUDY DESIGN: Human endometrial tissues were obtained from 18 patients aged 34-47 years undergoing hysterectomy for benign reasons. ESCs were cultured under hypoxic condition or treated with 17β-estradiol (E) and/or medroxyprogesterone acetate (MPA). The mRNA levels and production of ANGPT1, ANGPT2, and VEGF were assessed by real-time RT-PCR and ELISA, respectively. Analysis of hypoxia-inducible factor 1α (HIF-1α) and estrogen receptor α (ERα) protein levels were evaluated by Western blot analysis. RESULT(S): Hypoxia reduced the mRNA expression and protein production of ANGPT-1 in ESCs, whereas those of ANGPT2 were unaffected, resulting in an increase of the ANGPT2/ANGPT1 ratio. Hypoxia induced mRNA expression and protein production of VEGF. E simultaneously induced VEGF production and suppressed ANGPT1 production, resulting in an increase of the ANGPT2/ANGPT1 ratio. MPA or E+MPA reduced ANGPT2 production and sustained the levels of ANGPT1, resulting in a decrease of the ANGPT2/ANGPT1 ratio. With regard to the interaction of E and hypoxia, E did not affect the regulation of angiogenic factors, HIF-1α, and ERα under hypoxic conditions. CONCLUSIONS: Hypoxia and female sex hormones independently regulate the ANGPT2/ANGPT1 ratio and VEGF expression in human ESCs. These results may indicate a potential mechanism for hypoxia or female sex steroids influencing angiogenesis in the human endometrium.
European journal of obstetrics, gynecology, and reproductive biology 01/2013; · 1.97 Impact Factor
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ABSTRACT: OBJECTIVE: To investigate whether heart and neural crest derivatives expressed transcript 2 (HAND2) regulates fibulin-1 (FBLN1) expression during decidualization of human endometrial stromal cells (ESCs). DESIGN: In vitro experiment. SETTING: Research laboratory. PATIENT(S): Twenty-four patients undergoing hysterectomy for benign reasons. INTERVENTION(S): ESCs were cultured with various progestins (medroxyprogesterone acetate [MPA], norethisterone, levonorgestrel, dienogest, and P), E(2), dexamethasone, and/or 8-bromoadenosine 3', 5'-cyclic monophosphate (8-Br-cAMP). HAND2 and FBLN1 were silenced by small interfering RNA technology. MAIN OUTCOME MEASURE(S): HAND2 and FBLN1 expression levels were assessed by real-time polymerase chain reaction and Western blot analysis. RESULT(S): MPA or E(2) + MPA increased HAND2 mRNA levels in ESCs in a time- and dose-dependent manner, and this stimulatory effect was blocked by RU-486 (P receptor antagonist). HAND2 was increased by E(2) + MPA earlier than FBLN1. Simultaneous MPA and 8-Br-cAMP treatment synergistically enhanced HAND2 mRNA levels. P and all the progestins significantly increased HAND2 mRNA levels, whereas E(2), 8-Br-cAMP, or dexamethasone alone had no effect. Silencing of HAND2 expression significantly reduced FBLN1 expression, whereas FBLN1 silencing had no effect on HAND2 expression. CONCLUSION(S): These results suggest that progestin-induced HAND2 contributes to FBLN1 expression in human ESCs.
Fertility and sterility 10/2012; · 3.97 Impact Factor
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ABSTRACT: Shakuyaku-kanzo-to (SK) is a herbal medicine and is known to possess an antispasmodic effect on skeletal muscle and intestinal smooth muscle. However, it is unclear whether SK is effective in antagonizing uterine smooth muscle contractions. Herein, we investigated the effects of SK on smooth muscle contractions of human pregnant uterine samples.
We prepared myometrial strips from uterine tissues of pregnant women who underwent cesarean section for obstetrical indications, and examined the inhibitory effects of SK and its components, shakuyaku (S) and kanzo (K), on agonist-induced and spontaneous contractions in vitro. Oxytocin, prostaglandinF(2α) , and high KCl were utilized as agonists in this study.
SK inhibited agonist-induced and spontaneous contractions in a dose-dependent manner. Inhibition of SK on oxytocin-induced contractions occurred at a concentration of 100 µg/mL and reached maximum effect at a concentration of more than 1000 µg/mL. The half max inhibitory concentration of SK was approximately 440 µg/mL in oxytocin-induced contractions. SK at 1000 µg/mL completely inhibited the oxytocin- and prostaglandinF(2α) -induced contractions but not the high KCl-induced contractions. The inhibitory effects on agonist-induced contractions of K, but not S, matched those of SK.
These results suggest that the inhibitory effect of SK on smooth muscle contractions is due to K. The mechanism of the inhibitory effects of SK on oxytocin- and prostaglandinF(2α) -induced contractions may differ from that on KCl-induced contractions.
Journal of Obstetrics and Gynaecology Research 05/2012; 38(7):1004-10. · 0.94 Impact Factor
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ABSTRACT: To determine the concentrations of angiopoietin 1 (ANGPT1) and ANGPT2 in individual human preovulatory follicles in relation to their diameter or volume to clarify the role of these molecules in folliculogenesis.
Prospective study.
Research laboratory at Kansai Medical University, Osaka, Japan.
Twenty-three women undergoing IVF.
On the day of oocyte retrieval, serum samples and follicular fluid (FF) from individual follicles were collected. We analyzed 348 follicles.
ANGPT1 and ANGPT2 concentrations in FF and serum and oocyte recovery rates.
On average, ANGPT1 concentrations in FF were 150 times lower than those in serum, whereas ANGPT2 concentrations in FF were 8 times higher than those in serum. The concentrations of ANGPT1 in follicles with a diameter ≤17 mm were significantly higher than those in follicles with a diameter ≥18 mm. On the other hand, the concentrations of ANGPT2 in follicles with a diameter ≤17 mm were significantly lower than those in follicles with a diameter ≥18 mm. The ANGPT2/ANGPT1 ratio increased with enlargement of follicular diameter. ANGPT1 concentrations in FF decreased with follicular volume. ANGPT2 concentrations and the ANGPT2/ANGPT1 ratio in FF rose with follicular volume. The ANGPT2/ANGPT1 ratio in FF from the oocyte recovery group was significantly higher than that from the nonrecovery group.
Our data suggested that the change in ANGPT1 and ANGPT2 levels may be associated with follicular growth and angiogenesis during the preovulatory period.
Fertility and sterility 12/2011; 96(6):1378-83. · 3.97 Impact Factor
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ABSTRACT: Hypoxia of the human endometrium is a physiologic event occurring during the perimenstrual period and the local stimulus for angiogenesis. The aim of this study was to investigate the effects of hypoxic stress on the regulation of vascular endothelial growth factor (VEGF) and stromal cell-derived factor-1 (SDF-1/CXCL12), and the potential role of hypoxia-inducible factor-1α (HIF-1α) in the endometrium.
Human endometrial stromal cells (ESCs, n= 22 samples) were studied in vitro. ESCs were cultured under hypoxic and normoxic conditions and treated with cobalt chloride (CoCl₂; a hypoxia-mimicking agent) and/or echinomycin, a small-molecule inhibitor of HIF-1α activity. The mRNA levels and production of VEGF and SDF-1 were assessed by real-time PCR and ELISA, respectively. The HIF-1α protein levels were measured using western blot analysis.
Hypoxia simultaneously induced the expression of mRNA and production of VEGF and attenuated the expression and production of SDF-1 from ESCs in a time-dependent manner. Similar changes were observed in the ESCs after stimulation with CoCl₂ in a dose-dependent manner. CoCl₂ significantly induced the expression of HIF-1α protein, and its highest expression was observed at 6 h. Echinomycin inhibited hypoxia-induced VEGF production without affecting the HIF-1α protein level and cell toxicity and had no effect on SDF-1 secretion (P < 0.01).
Hypoxia simultaneously acts to increase VEGF via HIF-1α and to decrease SDF-1 in a HIF-1α-independent manner in ESCs. These results indicate a potential mechanism for the action of hypoxic conditions that could influence angiogenesis in the human endometrium.
Human Reproduction 11/2011; 27(2):523-30. · 4.47 Impact Factor
