Journal of Endocrinological Investigation

, Volume 32, Issue 5, pp 415–419 | Cite as

Modulating effect of leptin on basal and follicle stimulating hormone stimulated steroidogenesis in cultured human lutein granulosa cells

  • M. Karamouti
  • P. Kollia
  • A. Kallitsaris
  • N. Vamvakopoulos
  • G. Kollios
  • I. E. Messinis
Original Articles


Background: In vitro data have shown conflicting results in terms of the effect of leptin on granulosa cells steroidogenesis. Aim: The aim of the present study was to investigate the effect of low and high doses of leptin on basal and FSH-induced steroids secretion by human luteinized granulosa cells in culture. Materials and methods: Granulosa cells were obtained from normal women undergoing in vitro fertilization (IVF) treatment and were cultured in serum-free conditions for 72 h. A one-way analysis of variance design was set to study the effect of leptin on basal and FSH-induced steroidogenesis. Results: Leptin affected basal estradiol and progesterone secretion in a dose-related manner. In particular, leptin at low concentrations stimulated the secretion of estradiol (1 and 10 ng/ml) and progesterone (10 ng/ml), while at a high concentration (100 ng/ml) it suppressed the secretion of both steroids. A dose-related effect of leptin on FSH-induced steroidogenesis was not evident, since only the suppressive effect of the high concentration of leptin (100 ng/ml) reached statistical significance for both steroids. Conclusions: These results demonstrate that leptin affects the secretion of steroids in luteinized granulosa cells in a dose-dependent manner. Although a physiological role for leptin is possible, it is suggested that this protein is a mediator of negative rather than positive influential interactions on ovarian function that may compromise fertility.


Culture human leptin lutein granulosa cells ovarian steroidogenesis 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Mantzoros CS, Moschos SJ. Leptin: in search of role(s) in human physiology and pathophysiology. Clin Endocrinol 1998, 49: 551–67.CrossRefGoogle Scholar
  2. 2.
    Messinis IE, Milingos SD. Leptin in human reproduction. Hum Reprod Update 1999, 5: 52–63.PubMedCrossRefGoogle Scholar
  3. 3.
    Moschos S, Chan JL, Mantzoros CS. Leptin and reproduction: a review. Fertil Steril 2002, 77: 433–44.PubMedCrossRefGoogle Scholar
  4. 4.
    Cioffi J, Van Blerkom J, Antczak M, Shafer A, Wittmer S, Snodgrass R. The expression of leptin and its receptors in pre-ovulatory human follicles. Mol Hum Reprod 1997, 3: 467–72.PubMedCrossRefGoogle Scholar
  5. 5.
    Karlsson C, Lindell K, Svensson E, et al. Expression of functional leptin receptors in the human ovary. J Clin Endocrinol Metab 1997, 82: 4144–8.PubMedGoogle Scholar
  6. 6.
    Agarwal SK, Vogel K, Weitsman SR, Magoffin DA. Leptin antagonizes the insulin-like growth factor-I augmentation of steroidogenesis in granulosa cells and theca cells of the human ovary. J Clin Endocrinol Metab 1999, 84: 1072–6.PubMedGoogle Scholar
  7. 7.
    Löeffler S, Aust G, Koehler U, Spanel-Borowski K. Evidence of l eptin expression in normal and polycystic human ovaries. Mol Hum Reprod 2001, 12: 1143–9.CrossRefGoogle Scholar
  8. 8.
    Karamouti M, Kollia P, Karligiotou E, et al. Absence of leptin expression and secretion by human luteinised granulosa cells. J Mol Endocrinol 2003,31: 233–9.PubMedCrossRefGoogle Scholar
  9. 9.
    Zachow RJ, Magoffin DA. Direct intraovarian effects of leptin: impairment of the synergistic action of Insulin-like Growth Factor-I on Follicle-Stimulating Hormone-Dependent Estradiol-17B production by rat ovarian granulosa cells. Endocrinology 1997, 138: 847–50.PubMedGoogle Scholar
  10. 10.
    Govoni N, Galeati G, Castellani G, Tamanini C. Leptin concentrations in plasma and follicular fluid from prepubertal gilts as influenced by fasting, refeeding and insulin. Horm Metab Res 2005, 37: 152–8.PubMedCrossRefGoogle Scholar
  11. 11.
    Spicer LJ, Francisco CC. The adipose obese gene product, leptin: Evidence of a direct inhibitory role in ovarian function. Endocrinology 1997, 138: 3374–9.PubMedGoogle Scholar
  12. 12.
    Spicer LJ, Francisco CC. Adipose obese gene product, leptin, inhibits bovine ovarian theca cell steroidogenesis. Biol Reprod 1998, 58: 207–12.PubMedCrossRefGoogle Scholar
  13. 13.
    Brannian J, Zhao Y, McElroy M. Leptin inhibits gonadotrophin-stimulated granulosa cell progesterone production by antagonizing insulin action. Hum Reprod 1999, 14: 1445–8.PubMedCrossRefGoogle Scholar
  14. 14.
    Kitawaki J, Kusuki I, Koshiba H, Tsukamoto K, Honjo H. Leptin directly stimulates aromatase activity in human luteinised granulosa cells. Mol Hum Reprod 1999, 5: 708–13.PubMedCrossRefGoogle Scholar
  15. 15.
    Greisen S, Ledet T, Moller N, et al. Effects of leptin on basal and FSH stimulated steroidogenesis in human granulosa luteal cells. Acta Obstet Gynecol Scand 2000, 79: 931–5.PubMedCrossRefGoogle Scholar
  16. 16.
    Ghizzoni L, Barreca A, Mastorakos G, et al. Leptin inhibits steroid biosynthesis by human granulosa-lutein cells. Horm Metab Res 2001, 33: 323–8.PubMedCrossRefGoogle Scholar
  17. 17.
    Sirotkin AV, Mlyncek M, Kotwica J, Makarevich AV, Florkovicova I, Hetenyi L. Leptin directly controls secretory activity of human granulosa cells: possible interrelationship with the IF/IGFBP system. Horm Res 2005, 64: 190–202.CrossRefGoogle Scholar
  18. 18.
    Messinis IE, Milingos S, Zikopoulos K, Kollios G, Seferiadis K, Lolis D. Leptin concentrations in the follicular phase of spontaneous cycles and cycles superovulated with follicle stimulating hormone. Hum Reprod 1998, 13: 1152–6.PubMedCrossRefGoogle Scholar
  19. 19.
    Yong EL, Baird DT, Yates R, Teichert LE, Hillier SG. Hormonal regulation of the growth and steroidogenic function of human granulosa cells. J Clin Endocrinol Metab 1992, 74: 842–9.PubMedCrossRefGoogle Scholar
  20. 20.
    Foldesi I, Breckwoldt M, Neulen J. Oestradiol production by luteinised human granulosa cells: evidence of the stimulatory action of recombinant human follicle stimulating hormone. Hum Reprod 1998, 13: 1455–60.PubMedCrossRefGoogle Scholar
  21. 21.
    Lambert A, Harris SD, Knaggs P, Robertson WR. Improved FSH sensitization and aromatase assay in human granulosa-lutein cells. Mol Hum Reprod 2000, 6: 677–80.PubMedCrossRefGoogle Scholar
  22. 22.
    Laughlin GA, Yen SS. Hypoleptinemia in women athletes: absence of a diurnal rhythm with amenorrhea. J Clin Endocrinol Metab 1997, 82: 318–21.PubMedCrossRefGoogle Scholar
  23. 23.
    Shimizu H, Shimomura Y, Nakanishi Y, et al. Estrogen increases in vivo leptin production in rats and human subjects. J Endocrinol 1997, 154: 285–92.PubMedCrossRefGoogle Scholar
  24. 24.
    Messinis IE, Papageorgiou I, Milingos S, et al. Oestradiol plus progesterone treatment increases serum leptin concentrations in normal women. Hum Reprod 2001, 16: 1827–32.PubMedCrossRefGoogle Scholar
  25. 25.
    Karamouti M, Kollia P, Kallitsaris A, Vamvakopoulos N, Kollios G, Messinis IE. Growth hormone, insulin-like growth factor I, and leptin interaction in human cultured lutein granulosa cells steroidogenesis. Fertil Steril 2008, 90 (Suppl 4): 1444–50.PubMedCrossRefGoogle Scholar
  26. 26.
    Ruiz-Cortés ZT, Martel-Kennes Y, Gévry NY, Downey BR, Palin MF, Murphy BD. Biphasic effects of leptin in porcine granulosa cells. Biol Reprod 2003, 68: 789–96.PubMedCrossRefGoogle Scholar
  27. 27.
    McNatty KP, Makris A, DeGrazia C, Osathanondh R, Ryan KJ. The production of progesterone, androgens, and estrogens by granulosa cells, thecal tissue, and stromal tissue from human ovaries in vitro. J Clin Endocrinol Metab 1979, 49: 687–99.PubMedCrossRefGoogle Scholar
  28. 28.
    Selvaraj N, Israeli D, Amsterdam A. Partial sequencing of the rat steroidogenic acute regulatory protein message from immortalized granulosa cells: regulation by gonadotropins and isoproterenol. Mol Cell Endocrinol 1996, 123: 171–7.PubMedCrossRefGoogle Scholar
  29. 29.
    Zachow RJ, Weitsman SR, Magoffin DA. Leptin impairs the synergistic stimulation by transforming growth factor-beta of follicle-stimulating hormone-dependent aromatase activity and messenger ribonucleic acid expression in rat ovarian granulosa cells. Biol Reprod 1999, 61: 1104–9.PubMedCrossRefGoogle Scholar
  30. 30.
    Tsai EM, Yang CH, Chen SC, et al. Leptin affects pregnancy outcome of in vitro fertilization and steroidogenesis of human granulosa cells. J Assist Reprod Genet 2002, 19: 169–76.PubMedCentralPubMedCrossRefGoogle Scholar
  31. 31.
    Campbell BK, Scaramuzzi RJ, Webb R. Induction and maintenance of oestradiol and immunoreactive inhibin production with FSH by ovine granulosa cells cultured in serum-free media. J Reprod Fertil 1996, 106: 7–16.PubMedCrossRefGoogle Scholar
  32. 32.
    Holst N, Bertheussen K, Burhol PG, Forsdahl F. Medium-associated luteinization expressed as progesterone release in granulosaluteal cells isolated from patients undergoing in-vitro fertilization. Hum Reprod 1991, 6: 1343–8.PubMedGoogle Scholar
  33. 33.
    Carey PE, Stewart MW, Ashworth L, Taylor R. Effect of insulin therapy on plasma leptin and body weight in patients with type 2 diabetes. Horm Metab Res 2003, 35: 372–6.PubMedCrossRefGoogle Scholar
  34. 34.
    Mantzoros CS, Cramer DW, Liberman RF, Barbieri RL. Predictive value of serum and follicular fluid leptin concentrations during assisted reproductive cycles in normal women and in women with the polycystic ovarian syndrome. Hum Reprod 2002, 15: 539–44.CrossRefGoogle Scholar
  35. 35.
    De Placido G, Alviggi C, Clarizia R, et al. Intra-follicular leptin concentration as a predictive factor for in vitro oocyte fertilization in assisted reproductive techniques. J Endocrinol Invest 2006, 29: 719–26.Google Scholar

Copyright information

© Italian Society of Endocrinology (SIE) 2009

Authors and Affiliations

  • M. Karamouti
    • 1
  • P. Kollia
    • 2
  • A. Kallitsaris
    • 1
  • N. Vamvakopoulos
    • 2
  • G. Kollios
    • 3
  • I. E. Messinis
    • 1
  1. 1.Department of OB and GYN,Medical SchoolUniversity of ThessaliaLarissaGreece
  2. 2.Department of Biology, Medical SchoolUniversity of ThessaliaLarissa
  3. 3.Department of Biological ChemistryUniversity of LoanninaLoanninaGreece

Personalised recommendations