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Endocrinology Dec 2019Parturition is an essential process in placental mammals for giving birth to offspring. However, the molecular machineries of parturition are not fully understood. We...
Parturition is an essential process in placental mammals for giving birth to offspring. However, the molecular machineries of parturition are not fully understood. We investigated whether oxytocin plays a crucial role in the progress of parturition in cooperation with the prostaglandin F2α (PGF2α) receptor. We first examined alterations in the expression of uterine contraction-associated genes in uteri of oxytocin receptor-deficient mice (Oxtr-/-) during parturition. We found that induction of cyclooxygenase (COX)-2 and connexin 43 expression was impaired in Oxtr-/-, whereas that of PGF2α receptor expression was not. We next generated mice with double knockout of genes for the oxytocin receptor/oxytocin and PGF2α receptor (Oxtr-/-;Ptgfr-/- and Oxt-/-;Ptgfr-/-) and evaluated their parturition with Oxtr-/-, Oxt-/-, Ptgfr-/-, and wild-type mice. In Oxtr-/-;Ptgfr-/- and Oxt-/-;Ptgfr-/-, pregnancy rates were similar to those of other genotypes. However, normal parturition was not observed in Oxtr-/-;Ptgfr-/- or Oxt-/-;Ptgfr-/- because of persistent progesterone from the corpus luteum, as observed in Ptgfr-/-. We administered RU486, a progesterone antagonist, to Ptgfr-/-, Oxtr-/-;Ptgfr-/-, and Oxt-/-;Ptgfr-/- on gestation day 19. These mice were able to deliver a living first pup and the parturition onset was similar to that in Ptgfr-/-. Meanwhile, unlike Ptgfr-/-, ∼75% of Oxtr-/-;Ptgfr-/- and Oxt-/-;Ptgfr-/- administered RU486 remained in labor at 24 hours after the onset of parturition. All of the pups that experienced prolonged labor died. We thus revealed that the oxytocin receptor is an upstream regulator of COX-2 and connexin 43 in the uterus during parturition and that both oxytocin/oxytocin receptor and PGF2α receptor are major components for successful parturition.
Topics: Animals; Connexin 43; Cyclooxygenase 2; Female; Male; Mice; Mice, Knockout; Oxytocin; Parturition; Pregnancy; Progesterone; Receptors, Oxytocin; Receptors, Progesterone; Receptors, Prostaglandin; Uterus
PubMed: 31517984
DOI: 10.1210/en.2019-00499 -
Fertility and Sterility Apr 2015To determine whether berberine (BBR), a naturally occurring plant-derived alkaloid, inhibits the proliferation of human uterine leiomyoma (UtLM) cells.
OBJECTIVE
To determine whether berberine (BBR), a naturally occurring plant-derived alkaloid, inhibits the proliferation of human uterine leiomyoma (UtLM) cells.
DESIGN
Laboratory research.
SETTING
Laboratory.
PATIENT(S)
UtLM and normal human uterine smooth muscle (UtSMC) cell lines.
INTERVENTION(S)
Treatment with [1] BBR (10, 20, and 50 μM), [2] BBR (20 and 50 μM) and/or 17β-estradiol (E2; 10 and 100 nM), and [3] BBR (20 and 50 μM) and/or progesterone (P4; 10 and 100 nM) for 24 or 72 hours.
MAIN OUTCOME MEASURE(S)
Cell proliferation, cell cycle, apoptosis, and related genes expression were determined.
RESULT(S)
BBR inhibited UtLM cell proliferation by inducing G2/M cell cycle arrest and apoptosis. Cell cycle G2/M phase-related genes were altered by BBR treatment: the expression of cyclin A1, cyclin B1, and Cdk1 were down-regulated, while Cdk4, p21, and p53 were up-regulated. BBR-treated cells stained positively for annexin V and manifested increased BAX expression. E2- and P4-induced UtLM cell proliferation was blocked by BBR treatment. In marked contrast, even the highest concentration of BBR (50 μM) did not influence cell proliferation in UtSMC cells.
CONCLUSION(S)
BBR selectively inhibits cellular proliferation and blocks E2- and P4-induced cell proliferation in UtLM but not in normal UtSMC cells. In addition, BBR did not demonstrate cytotoxicity effects in normal human UtSMCs. Our results suggest BBR could be a potential therapeutic agent for the treatment of uterine leiomyoma.
Topics: Antineoplastic Agents, Phytogenic; Berberine; Cell Proliferation; Drug Antagonism; Estradiol; Estrogen Antagonists; Female; Gene Expression Regulation, Neoplastic; Humans; Leiomyoma; Progesterone; Tumor Cells, Cultured; Uterine Neoplasms
PubMed: 25682924
DOI: 10.1016/j.fertnstert.2015.01.010 -
Endocrinology May 2009LH and FSH play critical roles in mammalian reproduction by mediating steroidogenesis and gametogenesis in the gonad. Gonadal steroid hormone feedback to the...
LH and FSH play critical roles in mammalian reproduction by mediating steroidogenesis and gametogenesis in the gonad. Gonadal steroid hormone feedback to the hypothalamus and pituitary influences production of the gonadotropins. We previously demonstrated that progesterone differentially regulates the expression of the LH and FSH beta-subunits at the level of the gonadotrope: FSHbeta transcription is induced, whereas LHbeta is repressed. In this study, we investigated the mechanism of progesterone repression of LHbeta gene expression using immortalized gonadotrope-derived LbetaT2 cells. The progesterone suppression of both basal and GnRH-induced LHbeta gene expression occurs in a hormone- and receptor-dependent manner. Chromatin immunoprecipitation demonstrates that the hormone-bound progesterone receptor (PR) is recruited to the endogenous mouse LHbeta promoter. In addition, suppression requires both the amino-terminal and DNA-binding regions of PR. Furthermore, progesterone suppression does not require direct PR binding to the promoter, and, thus, PR is likely recruited to the promoter via indirect binding through other transcription factors. These data demonstrate that the molecular mechanism for progesterone action on the LHbeta promoter is distinct from FSHbeta, which involves direct PR binding to the promoter to produce activation. It also differs from androgen repression of LHbeta gene expression in that, rather than Sp1 or steroidogenic factor-1 elements, it requires elements within -300/-250 and -200/-150 that also contribute to basal expression of the LHbeta promoter. Altogether, our data indicate that progesterone feedback at the level of the pituitary gonadotrope is likely to play a key role in differential production of the gonadotropin genes.
Topics: Animals; Base Sequence; Binding Sites; Cells, Cultured; DNA; Drug Antagonism; Gene Expression Regulation; Gonadotrophs; Gonadotropin-Releasing Hormone; Humans; Luteinizing Hormone, beta Subunit; Mutation; Progesterone; Promoter Regions, Genetic; Protein Structure, Tertiary; Receptors, Progesterone; Sp1 Transcription Factor; Spodoptera
PubMed: 19106225
DOI: 10.1210/en.2008-1027 -
Fertility and Sterility Jun 1988Healthy, regularly menstruating women were treated with the antiprogesterone RU 486, Mifepristone (Roussel-Uclaf, Romainville, France) during the follicular phase of the... (Comparative Study)
Comparative Study
Healthy, regularly menstruating women were treated with the antiprogesterone RU 486, Mifepristone (Roussel-Uclaf, Romainville, France) during the follicular phase of the cycle. Three women were given 25 mg of RU 486 on days 1 to 14 of the cycle and five received 25 mg on days 1 to 21 of the cycle. Venous blood samples were collected three times per week during a control cycle and during one treatment cycle in each subject. Serum concentrations of estradiol (E2), progesterone (P), and RU 486 were determined by radioimmunoassays. No drug-related side effects and no spotting or bleeding during RU 486 treatment were observed. Menstrual bleeding was delayed by 8.7 +/- 3.8 days (mean +/- SD) after treatment over days 1 to 14 and by 12.6 +/- 3.2 days after treatment over days 1 to 21. During the treatment with RU 486, the serum concentrations of E2 remained low, indicating effective inhibition of folliculogenesis. After cessation of RU 486 treatment, serum E2 levels rose to similar values as in the control cycle, and subsequently serum P concentrations also reached ovulatory levels in six out of the eight volunteers. The results showed that the antiprogesterone RU 486 delayed folliculogenesis and luteinization even at low doses when given during the follicular phase of the menstrual cycle. It is speculated that this property of RU 486 could be utilized in the design of an estrogen-free combined oral contraceptives.
Topics: Adult; Contraceptives, Oral, Synthetic; Depression, Chemical; Estradiol; Estrenes; Female; Follicular Phase; Humans; Mifepristone; Ovarian Follicle; Ovulation; Progesterone; Radioimmunoassay; Time Factors
PubMed: 3371492
DOI: No ID Found -
The Journal of Clinical Endocrinology... May 2012Mifepristone is a glucocorticoid and progestin antagonist under investigation for the treatment of Cushing's syndrome. Mifepristone decreases high-density lipoprotein... (Randomized Controlled Trial)
Randomized Controlled Trial
CONTEXT
Mifepristone is a glucocorticoid and progestin antagonist under investigation for the treatment of Cushing's syndrome. Mifepristone decreases high-density lipoprotein (HDL) cholesterol (HDL-C) levels in treated patients, but the clinical significance of this is unclear because recent studies suggest that functional properties of HDL predict cardiovascular disease status better than does HDL-C concentration.
OBJECTIVE
The aim of the study was to characterize the impact of mifepristone administration on HDL particle concentration and function.
DESIGN AND SETTING
We conducted a double-blind, randomized, placebo-controlled trial at a single-site, clinical research center.
PARTICIPANTS
Thirty healthy postmenopausal female volunteers participated in the study.
INTERVENTION
Individuals were randomized to receive daily oral mifepristone (600 mg) or placebo for 6 wk.
MAIN OUTCOME MEASURES
We measured HDL-C, serum HDL particle concentration, and HDL-mediated cholesterol efflux by treatment group.
RESULTS
As expected, ACTH, cortisol, estradiol, and testosterone levels increased in the mifepristone group. Mifepristone treatment decreased HDL-C and HDL particle concentration by 26 and 25%, respectively, but did not alter pre-β HDL concentration. In contrast, the serum HDL-mediated cholesterol efflux decreased with mifepristone treatment by only 12%, resulting in an effective increase of the efflux capacity per HDL particle. No changes were observed in cholesterol ester transfer protein or lecithin:cholesterol acyltransferase activity.
CONCLUSIONS
Treatment with mifepristone reduced HDL-C, HDL particle concentration, and serum HDL cholesterol efflux in postmenopausal women. However, on a per particle basis, the efflux capacity of serum HDL increased. These observations support the concept that a decrease in HDL-C may not represent proportional impairment of HDL function.
Topics: Aged; Cholesterol, HDL; Double-Blind Method; Female; Hormone Antagonists; Humans; Lipoproteins, HDL; Middle Aged; Mifepristone; Postmenopause; Progesterone
PubMed: 22399518
DOI: 10.1210/jc.2011-2813 -
Journal of Applied Physiology... Mar 2004To determine the effect of estrogen and progesterone on plasma volume (PV) and extracellular fluid volume (ECFV), we suppressed endogenous estrogen and progesterone by... (Comparative Study)
Comparative Study
To determine the effect of estrogen and progesterone on plasma volume (PV) and extracellular fluid volume (ECFV), we suppressed endogenous estrogen and progesterone by using the gonadotropin-releasing hormone (GnRH) antagonist ganirelix acetate in seven healthy women (22 +/- 1 yr). Subjects were administered GnRH antagonist for 16 days. Beginning on day 5 of GnRH antagonist administration, subjects were administered estrogen (E(2)) for 11 days, and beginning on day 12 of GnRH antagonist administration, subjects added progesterone (E(2)-P(4)) for 4 days. On days 2, 9, and 16 of GnRH antagonist administration, we estimated ECFV (inulin washout), transcapillary escape rate of albumin (TER(alb)), and PV (Evans blue dye). Plasma E(2) concentration increased from 17.9 +/- 4.5 (GnRH antagonist) to 195.9 +/- 60.1 (E(2), P < 0.05) to 245.6 +/- 62.9 pg/ml (E(2)-P(4), P < 0.05). Compared with GnRH antagonist (1.3 +/- 0.5 ng/ml), plasma P(4) concentration was unchanged during E(2) (0.9 +/- 0.3 ng/ml) and increased to 9.4 +/- 3.1 ng/ml during E(2)-P(4) (P < 0.05). Both E(2) (44.1 +/- 3.1 ml/kg) and E(2)-P(4) (47.7 +/- 2.8 ml/kg) increased PV compared with GnRH antagonist (42.8 +/- 1.3 ml/kg, P < 0.05). Within-subjects TER(alb) was a strong negative predictor of PV (mean r = 0.92 +/- 0.03, P < 0.05), and TER(alb) was lowest during E(2)-P(4) (5.7 +/- 0.5, 4.1.0 +/- 1.1, and 2.8 +/- 0.9%/h, P < 0.05, for GnRH antagonist, E(2), and E(2)-P(4), respectively). ECFV was reduced during E(2) (227 +/- 31 ml/kg, P < 0.05) compared with both GnRH antagonist (291 +/- 37 ml/kg) and E(2)-P(4) (283 +/- 19 ml/kg). Thus the percentage of extracellular fluid in the plasma compartment increased to 21.0% (P < 0.05) during E(2) compared with GnRH antagonist (16.1%) and E(2)-P(4) (17.2%) administration. Thus E(2) increased PV via actions on the capillary endothelium to lower TER(alb) and favor intravascular water retention, whereas during E(2)-P(4) PV increased via the combined responses of ECFV expansion and lower TER(alb).
Topics: Adult; Estrogens; Extracellular Fluid; Female; Gonadotropin-Releasing Hormone; Humans; Luteal Phase; Plasma Volume; Progesterone
PubMed: 14660504
DOI: 10.1152/japplphysiol.01032.2003 -
American Journal of Physiology. Heart... Jan 2017Acute application of progesterone attenuates cardiac contraction, although the underlying mechanisms are unclear. We investigated whether progesterone modified...
UNLABELLED
Acute application of progesterone attenuates cardiac contraction, although the underlying mechanisms are unclear. We investigated whether progesterone modified contraction in isolated ventricular myocytes and identified the Ca handling mechanisms involved in female C57BL/6 mice (6-9 mo; sodium pentobarbital anesthesia). Cells were field-stimulated (4 Hz; 37°C) and exposed to progesterone (0.001-10.0 μM) or vehicle (35 min). Ca transients (fura-2) and cell shortening were recorded simultaneously. Maximal concentrations of progesterone inhibited peak contraction by 71.4% (IC = 160 ± 50 nM; n = 12) and slowed relaxation by 75.4%. By contrast, progesterone had no effect on amplitudes or time courses of underlying Ca transients. Progesterone (1 µM) also abbreviated action potential duration. When the duration of depolarization was controlled by voltage-clamp, progesterone attenuated contraction and slowed relaxation but did not affect Ca currents, Ca transients, sarcoplasmic reticulum (SR) content, or fractional release of SR Ca Actomyosin MgATPase activity was assayed in myofilaments from hearts perfused with progesterone (1 μM) or vehicle (35 min). While maximal responses to Ca were not affected by progesterone, myofilament Ca sensitivity was reduced (EC = 0.94 ± 0.01 µM for control, n = 7 vs. 1.13 ± 0.05 μM for progesterone, n = 6; P < 0.05) and progesterone increased phosphorylation of myosin binding protein C. The effects on contraction were inhibited by lonaprisan (progesterone receptor antagonist) and levosimendan (Ca sensitizer). Unlike results in females, progesterone had no effect on contraction or myofilament Ca sensitivity in age-matched male mice. These data indicate that progesterone reduces myofilament Ca sensitivity in female hearts, which may exacerbate manifestations of cardiovascular disease late in pregnancy when progesterone levels are high.
NEW & NOTEWORTHY
We investigated myocardial effects of acute application of progesterone. In females, but not males, progesterone attenuates and slows cardiomyocyte contraction with no effect on calcium transients. Progesterone also reduces myofilament calcium sensitivity in female hearts. This may adversely affect heart function, especially when serum progesterone levels are high in pregnancy.Listen to this article's corresponding podcast at https://ajpheart.podbean.com/e/acute-progesterone-modifies-cardiac-contraction/.
Topics: Animals; Calcium; Cardiotonic Agents; Carrier Proteins; Estrenes; Female; Heart Ventricles; Hydrazones; Male; Mice; Mice, Inbred C57BL; Myocardial Contraction; Myocytes, Cardiac; Myofibrils; Myosins; Phosphorylation; Progesterone; Progestins; Pyridazines; Receptors, Progesterone; Sarcoplasmic Reticulum; Simendan
PubMed: 27793852
DOI: 10.1152/ajpheart.00073.2016 -
Journal of Cellular and Molecular... Feb 2012The mechanisms of progesterone on endothelial cell motility are poorly investigated. Previously we showed that progesterone stimulated endothelial cell migration via the...
The mechanisms of progesterone on endothelial cell motility are poorly investigated. Previously we showed that progesterone stimulated endothelial cell migration via the activation of actin-binding protein moesin, leading to actin cytoskeleton remodelling and the formation of cell membrane structures required for cell movement. In this study, we investigated the effects of progesterone on the formation of focal adhesion complexes, which provide anchoring sites for cell movement. In cultured human umbilical endothelial cells, progesterone enhanced focal adhesion kinase (FAK) phosphorylation at Tyr(397) in a dose- and time-dependent manner. Several signalling inhibitors interfered with progesterone-induced FAK activation, including progesterone receptor (PR) antagonist ORG 31710, specific c-Src kinase inhibitor PP2, phosphatidylinosital-3 kinase (PI3K) inhibitor wortmannin as well as ρ-associated kinase (ROCK-2) inhibitor Y27632. It suggested that PR, c-Src, PI3K and ROCK-2 are implicated in this action. In line with this, we found that progesterone rapidly promoted c-Src/PI3K/Akt activity, which activated the small GTPase RhoA/ρ-associated kinase (ROCK-2) complex, resulting in FAK phosphorylation. In the presence of progesterone, endothelial cells displayed enhanced horizontal migration, which was reversed by small interfering RNAs abrogating FAK expression. In conclusion, progesterone promotes endothelial cell movement via the rapid regulation of FAK. These findings provide new information on the biological actions of progesterone on human endothelial cells that are relevant for vascular function.
Topics: Amides; Androstadienes; CSK Tyrosine-Protein Kinase; Cell Movement; Cells, Cultured; Endothelial Cells; Estrenes; Focal Adhesion Protein-Tyrosine Kinases; Focal Adhesions; Furans; Human Umbilical Vein Endothelial Cells; Humans; Phosphatidylinositol 3-Kinase; Phosphoinositide-3 Kinase Inhibitors; Phosphorylation; Progesterone; Protein-Tyrosine Kinases; Proto-Oncogene Proteins c-akt; Pyridines; Pyrimidines; RNA Interference; RNA, Small Interfering; Receptors, Progesterone; Signal Transduction; Wortmannin; rho-Associated Kinases; src-Family Kinases
PubMed: 21418517
DOI: 10.1111/j.1582-4934.2011.01305.x -
Fetal Diagnosis and Therapy 2012Mifepristone is a progesterone receptor antagonist widely used in obstetrics. The aim of the study was to focus on free corticotrophin-releasing hormone (CRH) and also... (Clinical Trial)
Clinical Trial
UNLABELLED
Mifepristone is a progesterone receptor antagonist widely used in obstetrics. The aim of the study was to focus on free corticotrophin-releasing hormone (CRH) and also describe modulation of adrenal and placental steroid hormone concentrations induced by mifepristone.
METHODS
Twenty-six women were enrolled in the study. They received mifepristone for termination of pregnancy. Maternal blood samples were retrieved before administration of mifepristone (600 mg) and 48 h after, just before induction of labor. Bound and free CRH levels were determined in maternal blood concomitantly with cortisol, estriol, progesterone and SDHEA levels. Also paired fetal cord blood samples were collected.
RESULTS
Maternal plasmatic CRH level did not change after mifepristone absorption but free CRH increased significantly (0.500 ± 0.326 vs. 0.388 ± 0.303 ng/ml, p = 0.040). A significant decrease of progesterone was observed (83.6 ± 49.3 vs. 95.6 ± 54.9 ng/ml, p = 0.001) with a lower progesterone/estriol ratio (26.9 ± 15.7 vs. 40.7 ± 31.1, p = 0.004). There was a strong association between maternal and fetal free CRH (r² = 0.675, p = 0.001), cortisol (r² = 0.570, p = 0.019), and positive but modest correlation for progesterone (r² = 0.341, p = 0.046) and estriol (r² = 0.379, p = 0.025) levels.
CONCLUSION
Mifepristone has an effect on free CRH level and changes the estriol-progesterone balance.
Topics: Abortifacient Agents, Steroidal; Abortion, Therapeutic; Adrenal Cortex; Adult; Algorithms; Corticotropin-Releasing Hormone; Dehydroepiandrosterone Sulfate; Estriol; Female; Fetal Blood; Humans; Hydrocortisone; Mifepristone; Paraventricular Hypothalamic Nucleus; Placenta; Pregnancy; Pregnancy Trimester, Third; Progesterone; Receptors, Progesterone
PubMed: 22759411
DOI: 10.1159/000338927 -
Proceedings of the National Academy of... Apr 1992Human progesterone receptors (hPRs) are phosphorylated at multiple serine residues, first in a basal step and then in a hormone-induced step. To determine whether...
Hormone-induced progesterone receptor phosphorylation consists of sequential DNA-independent and DNA-dependent stages: analysis with zinc finger mutants and the progesterone antagonist ZK98299.
Human progesterone receptors (hPRs) are phosphorylated at multiple serine residues, first in a basal step and then in a hormone-induced step. To determine whether hormone-induced phosphorylation precedes or follows the interaction of hPRs with DNA two strategies were used. (i) DNA binding was prevented or altered with site-specific mutants of the A form of hPR; (ii) DNA binding of wild-type hPR forms A and B was prevented with the progesterone antagonist ZK98299. Two hPRA mutants were constructed: DBDCys, which lacks a critical cysteine residue in the first zinc finger, and DBDsp, which is mutated at three discriminatory amino acids to change its DNA binding specificity from a progesterone response element to an estrogen response element. Receptors were transiently expressed in PR-negative cells and were intranuclear. DBDCys did not bind DNA in vitro and DBDsp bound only the estrogen response element. Transiently expressed hPRA and DBDsp showed the upward shift in electrophoretic mobility characteristic of hormone-induced phosphorylation; it was absent with DBDCys. Hormone-induced [32P] orthophosphate incorporation into transiently expressed DBDCys was reduced 60% compared to hPRA and DBDsp but was not eliminated. ZK98299 binds hPRs but prevents their interaction with DNA. Compared to R5020, the antagonist reduced phosphorylation of hPRB and hPRA in T47D breast cancer cells by 60% and totally prevented the mobility shift. We conclude that the hormone-induced phosphorylation of hPR includes DNA-independent and DNA-dependent stages and that only DNA-dependent sites contribute to the mobility shift.
Topics: Animals; Base Sequence; Chlorocebus aethiops; DNA; DNA-Binding Proteins; Gonanes; Humans; Ligands; Molecular Sequence Data; Mutagenesis, Site-Directed; Oligodeoxyribonucleotides; Phosphorylation; Phosphoserine; Progesterone; Receptors, Progesterone; Regulatory Sequences, Nucleic Acid; Structure-Activity Relationship; Transfection; Zinc Fingers
PubMed: 1557412
DOI: 10.1073/pnas.89.7.3050