Subscribe to RSS
DOI: 10.1055/s-2002-33891
Georg Thieme Verlag Stuttgart · New York
Effects of Recombinant Human FSH and hCG on the Expression of Interleukin-6 (IL-6) and Vascular Endothelial Growth Factor-A (VEGF-A) in Human Granulosa Lutein Cells and Stimulation of VEGF-A Expression by IL-6
Einfluss von rekombinantem hFSH und hCG auf die Expression von Interleukin-6 (IL-6) und Vascular Endothelial Growth Factor (VEGF) in humanen Granulosa Lutein Zellen und Stimulation der VEGF-A-Expression durch IL-6Publication History
Eingang Manuskript: 24 April 2002
Akzeptiert: 24 June 2002
Publication Date:
06 September 2002 (online)
![](https://www.thieme-connect.de/media/gebfra/200209/lookinside/thumbnails/10.1055-s-2002-33891-1.jpg)
Zusammenfassung
Einleitung
In der vorliegenden Studie wurde der Einfluss von Gonadotropinen auf die VEGF- und IL-6-Sekretion menschlicher Granulosazellen untersucht sowie der Einfluss von IL-6 auf die VEGF-Sekretion.
Methoden
Granulosazellen wurden aus der Follikelflüssigkeit von Patientinnen im Zusammenhang mit einer IVF-Behandlung gewonnen. Nach Stimulation der Zellen mit verschiedenen FSH- und hCG-Konzentrationen wurden die VEGF- und IL-6-Expressionen im Northern Blot gemessen. Die Bestimmung der VEGF- und IL-6-Proteinsekretion im Zellüberstand erfolgte durch ELISA. Die Zellen wurden darüber hinaus mit ansteigenden IL-6-Konzentrationen inkubiert und anschließend wurden die VEGF-A-Expression und ‐Sekretion ermittelt.
Ergebnisse
Die Stimulation von Granulosazellen sowohl mit rhFSH als auch mit hCG führte zu einer dosisabhängigen Steigerung der VEGF- und IL-6-Sekretion. Die Stimulation mit IL-6 führte zu einem Anstieg der VEGF-Sekretion.
Schlussfolgerung
Die VEGF- und IL-6-Sekretion menschlicher Granulosazellen wird durch Gonadotropine reguliert. IL-6 stimuliert die VEGF-Sekretion von Granulosazellen. Diese Mechanismen könnten zur Entwicklung des ovariellen Überstimulationssyndroms (OHSS) beitragen.
Abstract
Introduction
This study was performed to elucidate the effects of gonadotropins on the production of vascular endothelial growth factor-A (VEGF-A) and interleukin-6 (IL-6) and the effects of IL-6 on VEGF-A expression in human granulosa-lutein cells in vitro.
Methods
Granulosa cells (GC) were isolated from follicular aspirates of patients undergoing ovarian stimulation for in vitro-fertilization (IVF). Cells were cultured with or without recombinant human follicle stimulating hormone (rhFSH) and human chorionic gonadotropin (hCG) at various concentrations and cellular expression of VEGF-A and IL-6 was analysed on the mRNA-level by Northern analysis. VEGF-A- and IL-6 protein in cell supernatants was measured by enzyme-linked immunosorbent assay (ELISA). Furthermore GC were stimulated with increasing doses of recombinant human IL-6 (rhIL-6) and VEGF-A production of GC was measured.
Results
Stimulation of GC with either hCG or rhFSH led to a dose-dependent increase of VEGF-A- and IL-6 protein. Stimulation of GC with increasing doses of rhIL-6 led to a dose-dependent increase of VEGF-A protein.
Conclusion
These findings suggest a gonadotropin dependent expression of VEGF and IL-6 in human GC. Furthermore IL-6 stimulates VEGF-A-production of GC. These interactions may be of pathophysiological significance for the development of OHSS.
References
- 1 Adashi E Y. The potential relevance of cytokines to ovarian physiology: The emergent role of resident ovarian cells in white blood cell series. Endocrinol Rev. 1990; 11 454-464
- 2 Akira S, Hirano T, Taga T, Kishimoto T. Biology of multifunctional cytokines: IL-6 and related molecules (IL-1 and TNF). FASEB J. 1990; 4 2860-2867
- 3 Gorospe W C, Hughes F M, Spanelo B I. Interleukin-6: effects on and production by rat granulosa cells in vitro. Endocrinology. 1992; 130 1750-1752
- 4 Gorospe W C, Spangelo B L. Interleukin-6 production by rat granulosa cells in vitro effects of cytokines, follicle-stimulating hormone, and cyclic 3′, 5′-adenosine monoposphate. Biol Reprod. 1993; 48 538-543
- 5 Loret de Mola J R, Flores J P, Baumgardner G P, Goldfarb J M, Gindlesperger V, Friedlander M A. Elevated interleukin-6 levels in the ovarian hyperstimulation syndrome: Ovarian immunohistochemical localization of interleukin-6 signal. Obstet Gynecol. 1996; 87 581-587
- 6 Mikuni M. Effect of interleukin-2 and interleukin-6 on ovary in the ovulatory period-establishment of the new ovarian perfusion system and influence of interleukins on ovulation rate and steroid secretion. Hokkaido Igaku Zasshi. 1995; 70 561-572
- 7 Ferrara N, Henzel W J. Pituitary follicular cells secrete a novel heparin-binding growth factor specific for vascular endothelial cells. Biochem Biophys Res Commun. 1989; 161 851-858
- 8 Ferrara N, Houck K A, Jakamen L B. et al . The vascular endothelial growth factor family of polypeptides. J Cell Biochem. 1991; 47 211-218
- 9 Connolly D T. Vascular permeability factor: a unique regulator of blood vessel function. J Cell Biochem. 1991; 47 219-233
- 10 Ferrara N, Davis-Symth T. The biology of vascular endothelial growth factor. Endocrine Rev. 1997; 18 4-25
- 11 Hazzard T M, Stouffer R L. Angiogenesis in ovarian follicular and luteal development. Balliére's Clin Obstet & Gynaecol. 2000; 14 883-900
- 12 Shweiki D, Itin A, Neufeld G. Patterns of expression of vascular endothelial growth factor (VEGF) and (VEGF) receptors in mice suggest a role in hormonally regulated angiogenesis. J Clin Invest. 1993; 91 2235-2243
- 13 Goldsman M P, Pedram A, Dominguez C E. Increased capillary permeability induced by human follicular fluid: a hypothesis for an ovarian origin of the hyperstimulation syndrome. Fertil Steril. 1995; 63 268-272
- 14 McClure N, Healy D L, Rogers P AW. Vascular endothelial growth factor as capillary permeability agent in ovarian hyperstimulation syndrome. Lancet. 1994; 344 235-236
- 15 Yan Z, Weich H A, Bernart W, Breckwoldt M, Neulen J. Vascular endothelial growth factor (VEGF) messenger ribonucleic acid (mRNA) expression in luteinized human granulosa cells in vitro. J Clin Endocrinol Metab. 1993; 77 1723-1725
- 16 Neulen M, Yan Z, Racezek S, Weindel K, Keck C, Weich H A, Marme D, Breckwoldt M. Human chorionic gonadotropin-dependent expression of vascular endothalial growth factor/vascular permeability factor in human granulosa cells: importance in ovarian hyperstimulation syndrome. J Clin Endocrinol Metab. 1995; 80 1967-1971
- 17 Doldi N, Bassan M, Fusi F M, Ferrari A. In controlled ovarian hyperstimulation, steroid production, oocyte retrieval, and pregnancy rate correlate with gene expession of vascular endothelial growth factor. J Assist Reprod Genet. 1997; 14 589-592
- 18 Buyalos R P, Watson J M, Martinez-Maza O. Detection of IL-6 in human follicular fluid. Fertil Steril. 1992; 57 1230-1234
- 19 Friedlander M A, Loret de Mola J R, Goldfarb J M. Elevated levels of IL-6 in ascites and serum from women with ovarian hyperstimulation syndrome. Fertil Steril. 1993; 60 826-833
- 20 Orvieto R, Voliovitch I, Fishman P, Ben-Rafael Z. Interleukin-6 and ovarian hyperstimulation syndrome: a pilot study. Hum Reprod. 1995; 10 24-27
- 21 Loret de Mola J R, Arredondo-Soberon F, Randle C P, Tureck R T, Friedlander M A. Markedly elevated cytokines in pleural effusion during the ovulation hyperstimulation syndrome: transudate or ascites?. Fertil Steril. 1997; 67 780-782
- 22 Aboulghar M A, Mansour R T, Serour G I, El Helw B A, Shaarawy M. Elevated levels of interleukin-2, soluble interleukin-2 receptor alpha, interleukin-6, soluble interleukin-6 receptor and vascular endothelial growth factor in serum and ascitic fluid of patients with severe ovarian hyperstimulation syndrome. Eur J Obstet Gynecol Reprod Biol. 1999; 87 81-85
- 23 Chen C-D, Wu M-Y, Chen H-F, Chen S-U, Ho H-N, Yang Y-S. Relationships of serum pro-inflammatory cytokines and vascular endothelial growth factor with liver dysfunction in severe ovarian hyperstimulation syndrome. Hum Reprod. 2000; 15 66-71
- 24 Abramov Y, Schenker J G, Lewin A, Friedler S, Nilsman B, Barak V. Plasma inflammatory cytokines correlate to the ovarian hyperstimulation syndrome. Hum Reprod. 1996; 11 1381-1386
- 25 Geva E, Lessing J B, Lerner-Geba L, Azem F, Yovel I, Amit A. Elevated levels of interleukin-6 in follicular fluid at the time of oocyte retrieval for in vitro fertilization predict the development of early-form ovarian hyperstimulation syndrome. Fertil Steril. 1997; 68 133-137
- 26 Keck C, Rajabi Z, Pfeifer K, Bettendorf H, Brandstetter T, Breckwoldt M. Expression of interleukin-6 and interleukin-6 receptors in human granulosa lutein cells. Mol Human Reprod. 1998; 4 1071-1076
- 27 Beckmann M W, Polacek D, Seung L. Human ovarian granulosa cell culture determination of blood cell contamination and evaluation of possible culture purification steps. Fertil Steril. 1991; 56 881-887
- 28 Best C L, Griffin P M, Hill J A. Interferon gamma inhibits luteinized human granulosa cell steroid production in vitro. Am J Obstet Gynecol. 1995; 172 1505-1510
- 29 Machelon V, Gougeon A, Duquenne C, Testart J, Wallon C, Delattre R M, Emilie D, Galanaud P. Ovarian production of IL6 and its potential inhibitory effect on progesterone secretion in Cynomolgus fascicularis. CR ACAD SCI SER III. 1995; 318 1111-1118
- 30 Chen C-D, Chen H-F, Lu H-F, Chen S-U, Ho H-N, Yang Y-S. Value of serum and follicular fluid cytokine profile in the prediction of moderate to severe ovarian hyperstimulation syndrome. Hum Reprod. 2000; 15 1037-1042
- 31 Christenson L K, Stouffer R L. Follicle-stimulating hormone and luteinizing hormone/chorionic gonadotropin stimulation of vascular endothelial growth factor production by macaque granulosa cells from pre- and periovulatory follicles. J Clin Endocrinol Metab. 1997; 82 2135-2142
- 32 Yan Z, Neulen J, Razek S, Weich H A, Keck C, Grunwald K, Breckwoldt M. Vascular endothelial growth factor (VEGF)/vascular permeability factor (VPF) production by lutinized human granulosa cells in vitro; a paracrine signal in corpus luteum formation. Gynecol Endocrinol. 1998; 12 149-153
- 33 Cohen T, Nahari D, Cerem L W, Neufeld G, Levi B Z. Interleukin 6 induces the expression of vascular endothelial growth factor. J Biol Chem. 1996; 271 736-741
- 34 Mathur R S, Jenkins J M, Bansal A S. The possible role of the immune system in the aetipathogenesis of ovarian hyperstimulation syndrome. Hum Reprod. 1997; 12 2629-2634
PD Dr. C. Keck
Department of Obstetrics and Gynecology
Hugstetter Straße 55
79106 Freiburg
Germany
Email: CKECK@FRK.UKL.uni-freiburg.de