iturelix and Endometriosis

To evaluate the mechanism of action of recombinant human tumor necrosis factor (TNF)-binding protein-1 by assessing differential expression of messenger RNA (mRNA) for cytokines, matrix metalloproteinases, and growth and adhesion factors in baboons.. Analysis of gene expression in a prospective randomized study.. University Fertility Center.. In the in vivo study, 14 baboons were randomly and subcutaneously (SC) treated with either phosphate-buffered saline (PBS), GnRH antagonist, or recombinant human TNF-binding protein-1 at the time of induction. In the ex vivo study, 4 baboons were treated by menstrual endometrium that had been incubated randomly with either PBS or recombinant human TNF-binding protein-1 before intrapelvic injection.. In the in vivo study, analysis of 11 endometrial and 10 endometriosis biopsies included either PBS (n = 5), GnRH antagonist (n = 8), or recombinant human TNF-binding protein-1 (n = 8). In the ex vivo study, 2 endometrial and 4 endometriosis biopsies were analyzed from 4 baboons.. The mRNA expression of TNF-alpha, IL-8, IL-6, transforming growth factor-beta (TGF-beta), vascular endothelial growth factor, intercellular adhesion molecule-1, matrix metalloproteinase-1, and regulated on activation, normal T-cell expressed and secreted were investigated using real-time reverse transcriptase-polymer chain reaction (PCR).. TGF-beta mRNA expression was decreased in endometriotic lesions from baboons treated with recombinant human TNF-binding protein-1 when compared with the placebo group.. Except TGF-beta, mRNA expression of inflammatory cytokines and adhesion/growth factors is not affected in endometrial and endometriosis biopsies from baboons after induction of endometriosis combined with either systemic injection of recombinant human TNF-binding or GnRH antagonist or ex vivo treatment with recombinant human TNF-binding protein-1. Further studies are needed to elucidate the mode of action on how inhibition of TNF-alpha activity prevents the development of endometriosis.

Topics: Animals; Biopsy; Carrier Proteins; Cell Adhesion Molecules; Cytokines; Endometriosis; Endometrium; Female; Gene Expression Regulation; Gonadotropin-Releasing Hormone; Hormone Antagonists; Humans; Inflammation; Intercellular Signaling Peptides and Proteins; Oligopeptides; Papio; Placebos; Recombinant Proteins; RNA, Messenger

Our purpose was to evaluate the effect of the GnRH agonist (GnRHa), leuprolide acetate (LA), and the GnRH antagonist (GnRHant), Antide, on apoptosis and expression of apoptosis-related proteins in endometrial epithelial cell (EEC) cultures from patients with endometriosis and controls (infertile women without endometriosis).. Biopsy specimens of eutopic endometrium were obtained from 22 patients with endometriosis and from 14 women that served as controls. Apoptosis was examined in EEC after incubation with LA and Antide. Bax, Bcl-2, Fas and FasL expression was evaluated after exposure to LA, Antide or a combination of both. The percentage of apoptotic cells (%ApC) was assessed by the acridine orange-ethidium bromide technique, and protein expression was evaluated by western blot and immunocytochemistry.. LA 100 and 1000 ng/ml increased the %ApC in EEC from patients with endometriosis (both P < 0.05) and controls (p < 0.05 and P < 0.01, respectively). Antide 10(-5) M increased the %ApC in EEC from patients with endometriosis and controls (P < 0.01). In EEC from women with endometriosis, Bax expression increased after treatment with LA, Antide and LA + Antide (P < 0.05, P < 0.001 and P < 0.001), whereas Bcl-2 expression decreased after exposure to LA and Antide (P < 0.001 and P < 0.01). FasL expression increased after LA, Antide and LA + Antide treatments (P < 0.01, P < 0.001 and P < 0.01). No significant changes were observed on Fas expression.. GnRH analogues enhanced apoptosis in EEC, and this was accompanied by an increase in expression of the pro-apoptotic proteins Bax and FasL and a decrease in expression of the anti-apoptotic protein Bcl-2.

Topics: Apoptosis; bcl-2-Associated X Protein; Cells, Cultured; Endometriosis; Endometrium; Epithelial Cells; Fas Ligand Protein; fas Receptor; Female; Gene Expression Regulation; Gonadotropin-Releasing Hormone; Humans; Immunohistochemistry; Infertility, Female; Leuprolide; Oligopeptides; Proto-Oncogene Proteins c-bcl-2

The aim of the present study was to evaluate the effect of GnRH analogues on the in-vitro eutopic endometrial cell apoptosis and release of interleukin-1beta (IL-1beta) and vascular endothelial growth factor (VEGF).. Biopsy specimens of eutopic endometrium obtained from 16 women with untreated endometriosis and 14 controls were studied. Apoptosis, IL-1beta and VEGF release were evaluated in epithelial endometrial cell cultures after incubation with leuprolide acetate (LA) as GnRH agonist, antide as GnRH antagonist, and a combination of both. The percentage of apoptotic cells was evaluated by the acridine orange-ethidium bromide technique, and IL-1beta and VEGF concentrations were assessed by using commercial enzyme-linked immunosorbent assay (ELISA) kits.. We found that LA (100 ng/ml) enhanced apoptosis in endometrial cell cultures from endometriosis patients and controls and this effect was reversed by antide at 10(-7) mol/l. IL-1beta and VEGF release was downregulated by LA in cultures from controls and endometriosis patients. The addition of antide 10(-7) mol/l reversed this inhibition. Endometrial cultures treated with antide at 10(-7) mol/l did not show any significant effects compared with basal conditions.. GnRH agonists appear to have a direct effect in endometrial cells cultures, by enhancing the percentage of apoptotic cells and decreasing the release of pro-mitogenic cytokines such as IL-1beta and VEGF.

Topics: Apoptosis; Case-Control Studies; Cells, Cultured; Endometriosis; Endometrium; Female; Gonadotropin-Releasing Hormone; Humans; Interleukin-1; Leuprolide; Oligopeptides; Vascular Endothelial Growth Factor A

There is growing evidence that suggests a direct action of gonadotropin-releasing hormone agonist (GnRH-a) on endometrial growth. Consequently, our purpose was to evaluate the effect of GnRH-a on in vitro eutopic endometrial cell growth and apoptosis.. Prospective study.. Research institute and clinical fertility center.. Sixteen women with untreated endometriosis and 14 controls.. Biopsy specimens of eutopic endometrium were obtained from all subjects. Apoptosis and cell proliferation were examined in epithelial endometrial cell cultures after incubation with leuprolide acetate (LA), antide, and a combination of both.. The percentage of apoptotic cells was evaluated by the acridine orange-ethidium bromide technique; cell proliferation was assessed by (3)H-thymidine incorporation.. Leuprolide acetate (LA) (100 ng/mL) enhanced apoptosis in endometrial cultures from patients with endometriosis and controls, and this effect was reversed by antide 10(-7)M. Cell proliferation was down-regulated by LA at 1, 10, and 100 ng/mL in cultures from women without and with endometriosis. The addition of antide 10(-7)M reversed this inhibition.. GnRH-a appears to have a direct effect by enhancing the apoptotic index and decreasing the cell proliferation in endometrial cells.

Topics: Acridine Orange; Apoptosis; Biopsy; Cell Division; Endometriosis; Epithelial Cells; Female; Fertility Agents, Female; Gonadotropin-Releasing Hormone; Hormone Antagonists; Humans; Leuprolide; Oligopeptides; Prospective Studies; Thymidine; Transforming Growth Factor beta

Recurrent endometriosis in women is difficult to study because of the ethical consideration of performing repeated surgeries. Previously in the rat model we described therapeutic regression of endometriosis with the gonadotropin-releasing hormone antagonist antide. Presently we report the spontaneous and steroid-induced recurrence of endometriosis after withdrawal from antide therapy. Rats with endometriosis received antide or vehicle on days 0 (proestrus), 3, 6, and 9 and were killed on days 0, 6, 12, 18, 24, 30, and 42 (n = 4 antide-treated and 4 vehicle-treated rats killed per day). Additional antide-treated rats (n = 4 per treatment) received estrogen, progesterone, both estrogen and progesterone, cholesterol, and no steroid on day 9 and were killed on day 12. Antide significantly suppressed endometriotic implant size on days 12, 18, and 24. However, implant size spontaneously returned to pretreatment values by day 30. Administration of steroids on day 9 elicited regrowth of antide-suppressed endometriosis (estrogen plus progesterone greater than estrogen, progesterone, or cholesterol greater than no steroid) by day 12. This resilience of endometriosis offers an explanation for treatment failure and recurrence of the disease in women.

Topics: Animals; Cholesterol; Endometriosis; Estradiol; Estrogens; Female; Gonadotropin-Releasing Hormone; Oligopeptides; Progesterone; Rats; Rats, Inbred Strains; Recurrence

Steroids modulate the secretory activity of the uterus, but little is known of their effect on ectopic endometrium protein synthesis and secretion. We utilized two-dimensional electrophoresis to visualize proteins produced by the uteri and endometriotic implants of both steroid-treated and reproductively cyclic rats with and without surgically induced endometriosis. Of the greater than 300 proteins visualized, only the uterine cultures from progesterone (P)-stimulated, estrogen-suppressed rats contained a distinctive glycoprotein (P-induced uterine protein-1; molecular weight [Mr] 70,000; isoelectric point [pI] 5.7). This protein was not detected in any of the endometriotic implant cultures. Progesterone-induced uterine protein-1 could play a role in luteal or endometrial physiology and may be valuable in assessing endometrial function. The aberrant secretory behavior of the ectopic endometrium suggests a possible involvement in the reproductive dysfunction associated with endometriosis.

Topics: Animals; Cells, Cultured; Electrophoresis, Gel, Two-Dimensional; Endometriosis; Estradiol; Estrus; Female; Glycoproteins; Gonadotropin-Releasing Hormone; Oligopeptides; Progesterone; Rats; Rats, Inbred Strains; Reference Values; Transplantation, Autologous; Uterus

Adverse effects of the GnRH antagonist Antide on folliculogenesis and fertility were noted when we were evaluating the therapeutic value of Antide on endometriosis in a rat model. Cyclic rats with (n = 56) and without (n = 18) surgically induced endometriosis received Antide (2 mg/kg) or vehicle at noon on days 0 (proestrus), 3, 6, and 9. Rats were killed at noon on days 0, 6, 12, 18, 24, 30, 42, and 165. The number of antral follicles and the number of atretic antral follicles evaluated did not differ (P greater than 0.05) between endometriosis and control rats. Antide-treated rats had more (P less than 0.05 ) atretic antral follicles (64.7%) than vehicle-treated rats (15.3%). Antide-treated rat ovaries contained fewer corpora lutea than those of vehicle-treated rats on days 6, 12, and 18. No corpora lutea were found in Antide-treated rat ovaries after day 18. The abnormal ovarian morphology of the Antide-treated rats persisted for the duration of the project (165 days). Fertility (rats without endometriosis) was assessed by mating vehicle-and Antide-treated rats at spontaneous proestrus (n = 8) for eight posttreatment cycles as well as after follicular stimulation (n = 2). All vehicle-treated and no Antide-treated rats became pregnant. No oocytes were found in the oviducts of Antide-treated rats after eight cycles, indicating that ovulation had not occurred. The serum FSH and estradiol concentrations in the rats treated with Antide were lower (P less than 0.05) on days 6, 12, and 18, but rose to values equal to (days 24 and 30) or greater than (day 42) those in vehicle-treated rats. Serum progesterone levels in rats treated with Antide were lower (P less than 0.05) than those in vehicle-treated rats on all days tested. In conclusion, at a dosage sufficient to suppress reproductive cyclicity (and elicit the regression of endometriosis), Antide also caused long term follicular atresia and infertility.

Topics: Animals; Endometriosis; Estradiol; Estrus; Female; Follicle Stimulating Hormone; Follicular Atresia; Follicular Phase; Gonadotropin-Releasing Hormone; Infertility, Female; Oligopeptides; Ovarian Follicle; Ovary; Rats; Rats, Inbred Strains

Topics: Animals; Disease Models, Animal; Drug Evaluation, Preclinical; Endometriosis; Estradiol; Estrus; Female; Gonadotropin-Releasing Hormone; Oligopeptides; Rats; Rats, Inbred Strains