Effects of IL-6 on steroidogenesis of human granulosa cells in vitro

Nouri, Mohammad; Ghaffari, Maerefat; Salmasi, Ali; Farzadi, Laeya; Ghasemzadeh, Alieh; Lu, Sh; Mettler, Liza L

Effects of IL-6 on steroidogenesis of human granulosa cells in vitro

Vol. 2, Issue 1, / January-March 2001
(Original Article, pages 40-49)

Mohammad Nouri: - Biochemistry Department, Tabriz Univarsity of Medical Sciences, Tabriz, Iran
Maerefat Ghaffari: 1- Biochemistry Department, Tabriz Univarsity of Medical Sciences, Tehran, Iran; 2- Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran
Ali Salmasi: - Infertility Center, Kiel University, Kiel, Germany
Laeya Farzadi: - Department of Obstet . and Gynecol . of Tabriz University of Medical Science, Tabriz, Iran
Alieh Ghasemzadeh: - Department of Obstet . and Gynecol . of Tabriz University of Medical Science, Tabriz, Iran
Sh Lu: - Infertility Center, Kiel University, Kiel, Germany
Liza L Mettler: - Obs & Gyn Department of Keil University, Kiel, Germany

Received: 11/28/2000 Accepted: 3/1/2001 - Publisher : Avicenna Research Institute

Other Format

Abstract

Recent studies suggest that in addition to gonadotropins, immunological factors, such as cytokines play an important role in production of steroid hormones. The purpose of this study was to examine effects of IL-6 on basal and FSH stimulated secretions of estradiol and progesterone in the presence of androstendione by human granulosa cells (GC) in vitro. Graunlosa cells were harvested at the time of follicular aspiration after ovarian hyperstimulation according to standard protocols with hMG from patients undergoing IVFET. The cells (2 x 104 viable cells per well) were cultured with HAM's F-10 without any supplements (control) or increasing concentrations of recombinant human (rh) IL-6, (8,16,32,64,128 pg/ml) added in the absence or presence of FSH (96 IU/ml). Media were collected after 24,48,72 and 96 hours at a 24h interval and estradiol and progesterone levels were measured by an enzyme immunoassay (EIA) with automated system. Results of this study showed that leuteinized GC in the absence of FSH and the presence of androgen was able to produce estradiol and progesterone in vitro. This production was significantly increased in the presence of FSH. Basal and FSH stimulated productions of estradiol were significantly (P<0.05) inhibited by increasing amounts of IL-6. Although this inhibitory effect on basal production of progesterone was not significant. IL-6 in a dose-dependent manner significantly (P<0.05) inhibited FSH stimulated production of progesterone by GC. These results suggest that IL-6 may play an important role in the production of estradiol and progesterone and any disorders in level of IL-6 may cause estradiol and progesterone release disturbances.

Keywords: Steroidogenesis, Human graunlosa cells, IL-6, Cytokine

To cite this article:

Nouri M, Ghaffari M, Salmasi A, Farzadi L, Ghasemzadeh A, Lu S, et al. Effects of IL-6 on steroidogenesis of human granulosa cells in vitro. J Reprod Infertil. 2001;2(1):40-49.

References

Armstrong D.T., Goff A.K., Dorrington J.H. Regulation of follicular estrogen biosynthesis. In Midgley, A.R. Jr and Sadler, W.A. Jr (eds) Ovarian Follicular Development and Function. Raven Press, New York. 1997; pp:169-182
Erickson G.F. Normal regulation of ovarian androgen production,. Semin. Reprod. Endocrinol. 1993; 11: 307-312
Karnitis VJ., Townson DH., Friedman C.I. et al. Recombinant human follicle-stimulating hormone stimulates multiple follicular growth, but minimal estrogen production in gonadotropin-releasing hormone antagonist-treated monkeys: examining the role of luteinizing hormone in follicular development and steroidogenesis. J Clin Endocrinol Metab. 1994; 79(1):91-7
Smyth C.D., Miro F., Howles C.M., et al. Effect of luteinizing hormone on follicle stimulating hormone-activated paracrine signalling in rat ovary. Hum Reprod. 1995; 10(1):33-9
Spangelo B.L., MacLeod R.M., Isakson P.C. Production of interleukin-6 by anterior pituitary cells in vitro. Endocrinol. 1990; 126(1):582-6
Van Snick J. Interleukin-6: an overview. Annu Rev Immunol. 1990; 8:253-78
Kishimoto T. The biology of interleukin-6. Blood. 1989; 74(1):1-10
Watson J.M., Sensintaffar J.L., Berek J.S., et al. Constitutive production of interleukin 6 by ovarian cancer cell lines and by primary ovarian tumor cultures. Cancer Res. 1990; 50(21):6959-65
Alpizar E., Spicer L.J. Effects of interleukin-6 on proliferation and follicle-stimulating hormoneinduced estradiol production by bovine granulose cells in vitro: dependence on size of follicle. Biol Reprod. 1994; 50(1):38-43
Breard E., Benhaim A., Feral C., et al. Rabbit ovarian production of interleukin-6 and its potential effects on gonadotropin-induced progesterone secretion in granulosa and theca cells.J Endocrinol. 1998; 159(3):479-87
Keck C., Rajabi Z., Pfeifer K., et al. Expression of interleukin-6 and interleukin-6 receptors in human granulosa lutein cells. Mol Hum Reprod. 1998; 4 (11): 1071-6
Figenschau Y., Sundsfjord J.A., Yousef M.I., et al. A simplified serum-free method for preparation and cultivation of human granulosaluteal cells. Hum Reprod. 1997; 12(3):523-31
Foldesi I., Breckwoldt M., Neulen J. production by luteinized human granulosa cells: evidence of the stimulatory action of recombinant human follicle stimulating hormone. Hum Reprod. 1998; 13(6):1455-60
Wood A.M., Lambert A., Hooper M.A., et al. Exogenous steroids and the control of oestradiol secretion by human granulosa-lutein cells by follicle stimulating hormone and insulin-like growth factor-I. Hum Reprod. 1994; 9(1):19-23
Bernhisel M.A., Holman J.F., Haney A.F., et al. Estrogen and progesterone production by granulosa cell monolayers derived from in vitro fertilization procedures: lack of evidence for modulation by androgen. J Clin Endocrinol Metab. 1987; 64(6):1251-6
Schipper I., Fauser B.C., Van Gaver E.B., et al. Development of a human granulosa cell culture model with follicle stimulating hormone responsiveness. Hum Reprod. 1993; 8(9):1380-6
Mason H.D., Mannaerts B., de Leeuw R., et al. Effects of recombinant human follicle stimulating hormone on cultured human granulosa cells: comparison with urinary gonadotrophins and actions in preovulatory follicles. Hum Reprod. 1993; 8(11):1823-7
Bergh C., Selleskog U., Hillensjo T. Recombinant human gonadotropins stimulate steroid and inhibin production in human granulose cells. Eur J Endocrinol. 1997; 136(6):617-23
Harlow C.R., Shaw H.J., Hillier S.G. Factors influencing follicle-stimulating hormoneresponsive steroidogenesis in marmoset granulose cells: effects of androgens and the stage of follicular maturity. Endocrinol. 1988; 122(6):2780-7
Hillier S.G., Tetsuka M., Fraser H.M. Location and developmental regulation of androgen receptor in primate ovary. Hum Reprod. 1997; 12(1):107-11
Whitman G.F., Luciano A.A., Maier D.B. Human chorionic gonadotropin localization and morphometric characterization of human granulosa-luteal cells obtained during in vitro fertilization cycles. Fertil Steril. 1989; 51(3):475-9
Mangan D.F., Wahl S.M. Differential regulation of human monocyte programmed cell death (apoptosis) by chemotactic factors and proinflammatory cytokines. J Immunol. 1991;15;147(10):3408-12
Machelon V., Emilie D., Lefevre A., et al. Interleukin-6 biosynthesis in human preovulatory follicles: some of its potential roles at ovulation. JClin Endocrinol Metab. 1994; 79(2):633-42
Pitzel L., Jarry H., Wuttke W. Effects and interactions of prostaglandin F2 alpha, oxytocin, and cytokines on steroidogenesis of porcine luteal cells. Endocrinol. 1993; 132(2):751-6
Gorospe W.C., Hughes F.M., Spangelo B.L. Interleukin-6: effects on and production by rat granulosa cells in vitro. Endocrinol. 1992; 130(3): 1750-2
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(4):561-72
Hughes F.M., Fong Y.Y. Gorospe W.C. Interleukin-6 stimulates apoptosis in FSHstimulated rat granulosa cells in vitro: development of an in vitro model. Endocrine. 1994; 2: 997-1002
Loret de Mola J.R., Flores J.P., Baumgardner G.P. et al. Elevated interleukin-6 levels in the ovarian hyperstimulation syndrome: ovarian immunohistochemical localization of interleukin-6 signal. Obstet Gynecol. 1996; 87(4):581-7
Chen C.D., Chen H.F., Lu H.F. et al. Value of serum and follicular fluid cytokine profile in the prediction of moderate to severe ovarian hyperstimulation syndrome. Hum Reprod. 2000; 15(5):1037-42
Garrido N., Navarro J., Remohi J., et al. Follicular hormonal environment and embryo quality in women with endometriosis. Hum Reprod Update. 2000; Feb;6 (1):67-74
Nouri M, Ghaffari M, Salmasi A, Ghasemzadeh A. Serum and follicular fluid IL-6 and sex steroid hormone levels and their correlation of undergoing IVF-ET with endometriosis and pregnancy rate in women. J Reprod Infertil. 2000;1(4):55-62.
Hillier S.G., Whitelaw P.F., Smyth C.D. Follicular oestrogen synthesis: the 'two-cell, twogonadotrophin' model revisited. Mol Cell Endocrinol. 1994;100(1-2):51-4.
Spangelo B.L., Isakson P.C., MacLeod R.M. Production of interleukin-6 by anterior pituitary cells is stimulated by increased intracellular adenosine 3',5'-monophosphate and vasoactive intestinal peptide. Endocrinol. 1990; 127(1):403-9.
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 monophosphate. Biol Reprod. 1993; 48(3):538-43.