The effects of cetrorelix and triptorelin on the viability and steroidogenesis of cultured human granulosa luteinized cells.

BACKGROUND We investigated the effects of the gonadotropin-releasing hormone (GnRH) agonist triptorelin as well the GnRH antagonist cetrorelix those of on the viability and steroidogenesis in human granulosa luteinized (hGL) cell cultures. MATERIALS AND METHODS The hGL cells were obtained from 34 women undergoing ovarian stimulation for IVF treatment. The cells were cultured for 48 h with or without 1 nM or 3 nM of cetrorelix or triptorelin in serum-free media. The cell viability was evaluated by the MTT [3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide] assay. The concentrations of estradiol and progesterone in culture supernatants were measured by ELISA. RESULTS Treatment with triptorelin slightly increased cell viability, whereas treatment with 3 nM cetrorelix led to a significant decrease. Estradiol concentrations were reduced with 3 nM triptorelin. Cultures treated with high-dose of either cetrorelix or triptorelin tended to secrete less progesterone than controls. CONCLUSION Cetrorelix significantly reduces the viability of hGL cells. Triptorelin and cetrorelix may have minor effects on steroidogenesis. These results suggest that GnRH analogues may influence ovarian functions.

[1]  D. Hardy,et al.  Gonadotropin releasing hormone antagonists suppress aromatase and anti-Müllerian hormone expression in human granulosa cells. , 2010, Fertility and sterility.

[2]  R. Ferriani,et al.  Leuprolide acetate reduces both in vivo and in vitro ovarian steroidogenesis in infertile women undergoing assisted reproduction , 2008, Steroids.

[3]  N. Nikolettos,et al.  Gonadotropin-Releasing Hormone in the Ovary , 2007, Reproductive Sciences.

[4]  E. Metzen,et al.  Gonadotropin-releasing hormone antagonists do not influence the secretion of steroid hormones but affect the secretion of vascular endothelial growth factor from human granulosa luteinized cell cultures. , 2006, Fertility and sterility.

[5]  M. Peluffo,et al.  Effect of gonadotropin-releasing hormone agonist and antagonist on proliferation and apoptosis of human luteinized granulosa cells. , 2006, Fertility and sterility.

[6]  P. Leung,et al.  Molecular biology of gonadotropin-releasing hormone (GnRH)-I, GnRH-II, and their receptors in humans. , 2005, Endocrine reviews.

[7]  E. Porcu,et al.  The effect of leuprorelin on steroidogenesis of human preovulatory granulosa cells in vitro , 1996, Journal of Assisted Reproduction and Genetics.

[8]  M. Hickey,et al.  The effect of leuprolide acetate on steroidogenesis by granulosa and theca cells in vitro , 1991, Journal of in Vitro Fertilization and Embryo Transfer.

[9]  D. Shapiro.  An overview of GnRH antagonists in infertility treatments. Introduction. , 2003, Fertility and sterility.

[10]  P. Leung,et al.  Potential role of gonadotrophin-releasing hormone (GnRH)-I and GnRH-II in the ovary and ovarian cancer. , 2003, Endocrine-related cancer.

[11]  A. Rigamonti,et al.  Mechanisms of action of GH. , 2003, Journal of endocrinological investigation.

[12]  M. Aboulghar,et al.  GnRH antagonist in assisted reproduction: a Cochrane review. , 2002, Human reproduction.

[13]  O. Ortmann,et al.  Gonadotrophin-releasing hormone (GnRH) and GnRH agonists: mechanisms of action. , 2002, Reproductive biomedicine online.

[14]  A. Abou-Setta,et al.  Gonadotrophin-releasing hormone antagonists for assisted conception. , 2006, The Cochrane database of systematic reviews.

[15]  O. Ortmann,et al.  Actions of gonadotropin-releasing hormone antagonists on steroidogenesis in human granulosa lutein cells. , 2001, European journal of endocrinology.

[16]  Claudio Chillik,et al.  The role of LHRH agonists and antagonists. , 2001, Reproductive biomedicine online.

[17]  J. Rabinovici,et al.  Effects of gonadotrophin-releasing hormone agonists on human ovarian steroid secretion in vivo and in vitro-results of a prospective, randomized in-vitro fertilization study. , 2000, Human reproduction.

[18]  T. Hillensjö,et al.  Is there a difference in the function of granulosa-luteal cells in patients undergoing in-vitro fertilization either with gonadotrophin-releasing hormone agonist or gonadotrophin-releasing hormone antagonist? , 1999, Human reproduction.

[19]  S. Pavlou,et al.  Gonadotropin-releasing hormone antagonist versus agonist administration in women undergoing controlled ovarian hyperstimulation: cycle performance and in vitro steroidogenesis of granulosa-lutein cells. , 1995, American journal of obstetrics and gynecology.

[20]  G. Ossenkoppele,et al.  A tetrazolium-based colorimetric MTT assay to quantitate human monocyte mediated cytotoxicity against leukemic cells from cell lines and patients with acute myeloid leukemia. , 1994, Journal of immunological methods.

[21]  H. Minaguchi,et al.  Direct effects of gonadotropin-releasing hormone on the ovary in rats and humans. , 1994, Hormone research.

[22]  J. Parinaud,et al.  Modulation of the steroidogenesis of cultured human granulosa-lutein cells by gonadotropin-releasing hormone analogs. , 1993, The Journal of clinical endocrinology and metabolism.

[23]  M. Tesone,et al.  Effect of a gonadotropin-releasing hormone agonist on luteinizing hormone receptors and steroidogenesis in ovarian cells. , 1993, Fertility and sterility.

[24]  J. Tarín,et al.  Steroidogenesis in vitro of human granulosa-luteal cells pretreated in vivo with two gonadotropin releasing hormone analogs employing different protocols. , 1992, Gynecological endocrinology : the official journal of the International Society of Gynecological Endocrinology.

[25]  A. Pellicer,et al.  Steroidogenesis in vitro of human granulosa-luteal cells pretreated in vivo with gonadotropin-releasing hormone analogs. , 1990, Fertility and sterility.

[26]  A. Isetta,et al.  MTT colorimetric assay for testing macrophage cytotoxic activity in vitro. , 1990, Journal of immunological methods.

[27]  J. Parinaud,et al.  Effect of a luteinizing hormone‐releasing hormone agonist (Buserelin) on steroidogenesis of cultured human preovulatory granulosa cell , 1989, Fertility and sterility.

[28]  W. Dodson,et al.  Leuprolide acetate: serum and follicular fluid concentrations and effects on human fertilization, embryo growth, and granulosa-lutein cell progesterone accumulation in vitro. , 1988, Fertility and sterility.

[29]  D. Gerlier,et al.  Use of MTT colorimetric assay to measure cell activation. , 1986, Journal of immunological methods.

[30]  J E Rivier,et al.  Gonadotropin-releasing hormone analog design. Structure-function studies toward the development of agonists and antagonists: rationale and perspective. , 1986, Endocrine reviews.

[31]  T. Mosmann.  Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. , 1983, Journal of immunological methods.

[32]  A. Schally,et al.  Isolation and properties of the FSH and LH-releasing hormone. , 1971, Biochemical and biophysical research communications.