Abstract
Purpose: To investigate the secretion of progesterone (P4) and corticosteroid binding globulin (CBG) by granulosa luteal cells (GC) as well as the mRNA levels of CBG and 3β hydroxisteroid dehydrogenase (3β HSD), in women with and without endometriosis in vivo and in vitro.
Methods: Prospective study in a private, university-affiliated assisted reproduction unit, including women with severe endometriosis (n = 14) or without the disease (n = 20) undergoing in vitro fertilization/intracytoplasmic sperm injection and embryo transfer. GC were obtained from each follicle aspirated, pooled for each patient, and follicular and blood contaminating leukocytes depleted through immunomagnetic purification. Secreted P4 and CBG, and mRNA for both 3β HSD and CBG were determined in vivo and in vitro using RIA and reverse transcription followed by competitive polymerase chain reaction (cRT-PCR).
Results: The pattern of expression of 3β HSD and CBG mRNAs in vivo and in vitro was similar in both groups. Also, GC from patients with endometriosis produced equal amounts of P4 and CBG than controls without the disease, either in freshly isolated cells or in 24-h cultures.
Conclusions: The GC function in terms of 3β HSD and CBG mRNA expression and P4/CBG secretion does not seem to be altered in patients with endometriosis in comparison with those without this condition.
Keywords: CBG, endometriosis, follicular environment, granulosa cells, 3β HSD
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REFERENCES
- 1.Garrido N, Navarro J, Remohí J, Simón C, Pellicer A. Follicular hormonal environment and embryo quality in women with endometriosis. Hum Reprod Update. 2000;6(1):67–74. doi: 10.1093/humupd/6.1.67. [DOI] [PubMed] [Google Scholar]
- 2.Simón C, Gutiérrez A, Vidal A, de los Santos MJ, Tarín JJ, Remohí J, Pellicer A. Outcome of patients with endometriosis in assisted reproduction: Results from in vitro fertilization and oocyte donation. Hum Reprod. 1994;9:725–729. doi: 10.1093/oxfordjournals.humrep.a138578. [DOI] [PubMed] [Google Scholar]
- 3.Pellicer A, Oliveira N, Gutierrez A, Remohí J, Cano A, Rui A, Simón C. Implantation in endometriosis: Lessons learned from IVF and oocyte donation. In: Spinola P, Coutinho EM, editors. In Progress in Endometriosis. Parthenon: Casterton-Hill; 1994. pp. 177–183. [Google Scholar]
- 4.Díaz I, Navarro J, Blasco L, Simon C, Pellicer A, Remohí J. Impact of stages III-IV of endometriosis on recipients of sibling oocytes. Matched case-control study. Fertil Steril. 2000;74(1):31–34. doi: 10.1016/s0015-0282(00)00570-7. [DOI] [PubMed] [Google Scholar]
- 5.Brizek CL, Schlaff S, Pellegrini VA, Frank JB, Worrilow KC. Increased incidence of aberrant morphological phenotypes in human embryogenesis—an association with endometriosis. J Assist Reprod Genet. 1996;12:106–112. doi: 10.1007/BF02211378. [DOI] [PubMed] [Google Scholar]
- 6.Pellicer A, Oliveira N, Ruiz A, Remohí J. S. Exploring the mechanisms of endometriosis-associated infertility: An analysis of embryo development and implantation in assisted reproduction. Hum Reprod. 1994;9:725–729. doi: 10.1093/humrep/10.suppl_2.91. [DOI] [PubMed] [Google Scholar]
- 7.Pellicer A, Valbuena D, Bauset C, Albert C, Bonilla-Musoles F. R. J. S. The follicular endocrine environment in stimulated cycles of women with endometriosis: Steroid levels and embryo quality. Fertil Steril. 1998;69:1135–1141. doi: 10.1016/s0015-0282(98)00085-5. [DOI] [PubMed] [Google Scholar]
- 8.Speroff L, Glass RH, Kase NG. Clinical Gynecologic Endocrinology and Infertility. Baltimore: Williams and Wilkins; 1973. [Google Scholar]
- 9.Andersen CY. Levels of steroid-binding proteins and steroids in human preovulatory follicle fluid and serum as predictors of success in in vitro fertilization-embryo transfer treatment. J Clin Endocrinol Metab. 1990;71(5):1375–1381. doi: 10.1210/jcem-71-5-1375. [DOI] [PubMed] [Google Scholar]
- 10.American Fertility Society Revised American Fertility Society Classification of endometriosis. Fertil Steril. 1985;43:351–352. doi: 10.1016/s0015-0282(16)48430-x. [DOI] [PubMed] [Google Scholar]
- 11.Beckmann MW, Polacek D, Seung L, Schreiber JR. Human ovarian granulosa cell culture: Determination of blood cell contamination and evaluation of possible culture purification steps. Fertil Steril. 1991;56(5):881–887. doi: 10.1016/s0015-0282(16)54659-7. [DOI] [PubMed] [Google Scholar]
- 12.Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidiniumthiocyanate-phenol-chloroform extraction. Anal Biochem. 1987;16:156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
- 13.Harlow C., Cahill DJ, Maile LA, Talbot W. M., Wardle PG, et al. Reduced pre-ovulatory granulosa cell steroidogenesis in women with endometriosis. J Clin Endocrinol Metab. 1996;81:426–429. doi: 10.1210/jcem.81.1.8550788. [DOI] [PubMed] [Google Scholar]
- 14.Cahill DJ, Wardle PG, Maile LA, Harlow C., Hull MGR. Pituitary-ovarian dysfunction as a cause for endometriosisassociated and unexplained infertility. Hum Reprod. 1995;10:3142–3146. doi: 10.1093/oxfordjournals.humrep.a135876. [DOI] [PubMed] [Google Scholar]
- 15.Norman RJ, Bonello N, Jasper MJ, Van der Hoek KH. Leukocytes: Essential cells in ovarian function and ovulation. Reprod Med Rev. 1997;6:97–111. [Google Scholar]
- 16.Skrzypack J. Morphology and steroidogenesis of cultured granulosa cells from endometrioidally changed ovaries. Exp Clin Endocrinol Diabetes. 1996;103(4):228–232. doi: 10.1055/s-0029-1211355. [DOI] [PubMed] [Google Scholar]
- 17.Adashi EY. The potential relevance of cytokines to ovarian physiology: The emerging role of resident ovarian cells of white blood cell series. Endocr Rev. 1990;11:454–456. doi: 10.1210/edrv-11-3-454. [DOI] [PubMed] [Google Scholar]
- 18.Garrido N, Albert C, Mercader A, Remohí J, Simón C, Pellicer A. Leukocyte subpopulations in ovulatory follicles in patients with endometriosis (abstract No 172) Hum Reprod. 1999;14:227. [Google Scholar]
- 19.Nisolle M, Donnez J. Peritoneal endometriosis, ovarian endometriosis, and adenomyotic nodules of the rectovaginal septum are three different entities. Fertil Steril. 1997;68(4):585–596. doi: 10.1016/s0015-0282(97)00191-x. [DOI] [PubMed] [Google Scholar]
- 20.Brosens I. Diagnosis of endometriosis. Semin Reprod Endocrinol. 1997;15(3):229–233. doi: 10.1055/s-2008-1068752. [DOI] [PubMed] [Google Scholar]
- 21.Toya M, Saito H, Ohta N, Saito T, Kaneko T, Hiroi M. Moderate and severe endometriosis is associated with alterations in the cell cycle of granulosa cells in patients undergoing in vitro fertilization and embryo transfer. Fertil Steril. 2000;73(2):344–350. doi: 10.1016/s0015-0282(99)00507-5. [DOI] [PubMed] [Google Scholar]
- 22.Nakahara K, Saito H, Saito T, Ito M, Ohta N, Takahashi T, Hiroi M. Ovarian fecundity in patients with endometriosis can be estimated by the incidence of apoptotic bodies. Fertil Steril. 1998;69(5):931–935. doi: 10.1016/s0015-0282(98)00038-7. [DOI] [PubMed] [Google Scholar]
- 23.Carlberg M, Nejaty J, Froysa B, Guan Y, Soder O, Berqvist A. Elevated expression of tumor necrosis factor alpha in cultured granulosa cells from women with endometriosis. Hum Reprod. 2000;15(6):1250–1255. doi: 10.1093/humrep/15.6.1250. [DOI] [PubMed] [Google Scholar]
- 24.Lachapelle MH, Hemmings R, Roy DC, Falcone T, Miron P. Flow cytometric evaluation of leukocyte subpopulations in the follicular fluids of infertile patients. Fertil Steril. 1996;65(6):1135–1140. doi: 10.1016/s0015-0282(16)58327-7. [DOI] [PubMed] [Google Scholar]
- 25.Garrido N, Albert C, Krussel JS, O'Connor J. No differences in expression, production and secretion of vascular endothelial growth factor (VEGF) and interleukin-6 (IL-6) by granulosa cells in women with endometriosis compared to those without the disease. Fertil Steril. 2001;76(3):568–575. doi: 10.1016/s0015-0282(01)01961-6. [DOI] [PubMed] [Google Scholar]