Comparison of the Effects of GHRP-6, Ghrelin Hormone Agonist, on human immature oocyte in Culture Medium in 48 Hours

Subject Areas : Journal of Animal Biology

Caspian Ostadian ¹ , Nasim Hayati ² , Ziba Zahiri ³ , Ahmad Hosseini ⁴

1 - Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran
2 - Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran
3 - Department of Obstetrics and Gynecology, Reproductive Health Research Center, Alzahra Hospital, School of Medicine, Guilan University of Medical Sciences, Rasht, Iran
4 - Department of Obstetrics and Gynecology, Reproductive Health Research Center, Alzahra Hospital, School of Medicine, Guilan University of Medical Sciences, Rasht, Iran

Received: 2024-02-17 Accepted : 2024-05-02 Published : 2024-08-06

Keywords: Immature oocytes, Intra cytoplasmic sperm injection, In vitro maturation, Ghrelin hormone, GHRP-6,

Abstract :

In vitro maturation (IVM) is a method by which immature oocytes mature in a culture medium and laboratory conditions. Therefore, the culture medium plays a crucial role in the final result of the IVF outcome. This study investigates the impact of six different doses of the Ghrelin hormone agonist GHRP-6 in a synthetic culture medium containing human tubal fluid with serum albumin (HTF+HSA) on the maturation rate of human immature oocytes (GV) within 24 and 48 hours. The oocytes were collected from women under 35 years old, referring to Mehr Infertility Center/ Rasht /Iran, due to non-female factor infertility, and immature oocytes, unsuitable for oocytes cytoplasmic injection, were donated to this project. 30 immature oocytes were used in all 6 experimental and control groups. The criterion of cell maturity and progress of meiosis was the formation of the first polar body after 24 and 48 hours. Fisher's exact test was used to examine statistically significant differences between the groups. The addition of 75 ng/ml of GHRP-6 to the HTF culture medium led to a maturation of 70% of GVs after 24 hours which was statistically significant in comparison with other groups (p < 0.05), but not significant at 48h which was 80% (p > 0.05). There was no significant difference between the 50 ng/ml and 25ng/ml groups and the control group. On the other hand, it was observed that increasing the dose of GHRP-6 in the culture medium led to stop growth and cell death. The results indicated that the Ghrelin hormone agonist with a concentration of 75 ng/ml in the culture medium of human tubule fluid containing human serum can accelerate the meiosis process and better development of oocytes.

References:

1. Bednarek M.A., Feighner S.D., Pong S.S., McKee K.K., Hreniuk D.L., Silva M.V., Heck J.V. 2000. Structure-function studies on the new growth hormone-releasing peptide, ghrelin: minimal sequence of ghrelin necessary for activation of growth hormone secretagogue receptor 1a. Journal of Medicinal Chemistry, 43(23):4370-4376.
2. Cekleniak N.A., Combelles C.M., Ganz D.A., Fung J., Albertini D.F., Racowsky C. 2001. A novel system for in vitro maturation of human oocytes. Fertility and Sterility, 75(6):1185-1193.
3. Chian R.C., Li J.H., Lim J.H., Yoshida H. 2023. IVM of human immature oocytes for infertility treatment and fertility preservation. Reproductive Medicine and Biology, 22(1):e12524.
4. Cowley M.A., Smith R.G., Diano S., Tschöp M., Pronchuk N., Grove K. L, Horvath T.L. 2003. The distribution and mechanism of action of ghrelin in the CNS demonstrates a novel hypothalamic circuit regulating energy homeostasis. Neuron, 37(4):649-661.
5. De Vos M., Grynberg M., Ho T.M., Yuan Y., Albertini D.F., Gilchrist R.B. 2021. Perspectives on the development and future of oocyte IVM in clinical practice. Journal of Assisted Reproduction and Genetics, 38(6):1265-1280.
6. Dovolou E., Messinis I. E., Periquesta E., Dafopoulos K., Gutierrez-Adan A., Amiridis G.S. 2014. Ghrelin accelerates in vitro maturation of bovine oocytes. Reproduction in Domestic Animals, 49(4):665-672.
7. Du C., Li H., Cao G., Wang X.C., Li C. 2010. Expression of the orexigenic peptide ghrelin and the type 1a growth hormone secretagogue receptor in sheep oocytes and pre-implantation embryos produced in vitro. Reproduction in Domestic Animals, 45(1):92-98.
8. Fesahat F., Dehghani Firouzabadi R., Faramarzi A., Khalili M.A. 2017. The effects of different types of media on in vitro maturation outcomes of human germinal vesicle oocytes retrieved in intracytoplasmic sperm injection cycles. Clinical and Experimental Reproductive Medicine, 44(2):79-84.
9. Gaytan F., Barreiro M. L., Chopin L. K., Herington A.C., Morales C., Pinilla L. Tena-Sempere M. 2003. Immuno localization of ghrelin and its functional receptor, the type 1a growth hormone secretagogue receptor, in the cyclic human ovary. Journal of Clinical Endocrinology and Metabolism, 88(2):879-887.
10. Kheradmand A., Roshangar L., Taati M., Sirotkin A.V. 2009. Morphometrical and intracellular changes in rat ovaries following chronic administration of ghrelin. Tissue Cell, 41(5):311-317.
11. Kojima M., Hosoda H., Date Y., Nakazato M., Matsuo H., Kangawa K. 1999. Ghrelin is a growth-hormone-releasing acylated peptide from stomach. Nature, 402(6762):656-660.
12. Kojima M., Kangawa K. 2005. Ghrelin: structure and function. Physiological Reviews, 85(2):495-522.
13. Li Y., Liu H., Yu Q., Liu H., Huang T., Zhao S., Zhao H. 2019. Growth Hormone Promotes in vitro Maturation of Human Oocytes. Frontiers in Endocrinology, 10:485.
14. Li Y., Pan P., Yuan P., Qiu Q., Yang D. 2016. Successful live birth in a woman with resistant ovary syndrome following in vitro maturation of oocytes. Journal of Ovarian Research, 9(1):54.
15. Lim C.T., Kola B., Grossman A., Korbonits M. 2011. The expression of ghrelin O-acyltransferase (GOAT) in human tissues. Endocrine Journal, 58(8):707-710.
16. Łukaszyk A., Rafińska L., Sawiński P., Kasprzak A., Olejniczak K., Ruciński M., Sowiński J. 2009. Immunohistochemical and hybridocytochemical study on ghrelin signalling in the rat seminiferous epithelium. Folia Histochemistry and Cytobiology, 47(3):415-423.
17. Mabudi H., Jamili S., Majd N.E., Vosoughi G., Fatemi, M.R., Rashed S. 2011. The effects of ghrelin on ovary histology in Barbus sharpeyi. Journal of Animal Physiology and Animal Nutrition, 95(5):599-602.
18. O'Brien M., Earley P., Morrison J.J., Smith T.J. 2010. Ghrelin in the human myometrium. Reproductive Biology and Endocrinology, 8(55):1-11.
19. Rak A., Gregoraszczuk E.L. 2008. Modulatory effect of ghrelin in prepubertal porcine ovarian follicles. Journal of Physiology and Pharmacology, 59(4):781-793.
20. Sirini M.A., Anchordoquy J.M., Anchordoquy J.P., Pascua A.M., Nikoloff N., Carranza A, Furnus C.C. 2017. The presence of acylated ghrelin during in vitro maturation of bovine oocytes induces cumulus cell DNA damage and apoptosis, and impairs early embryo development. Zygote, 25(5):601-611.
21. Sirini M.A., Anchordoquy J.P., Quintana S., Furnus C., Relling A.E., Anchordoquy J.M. 2019. Expression of Ghrelin and Its Receptor mRNA in Bovine Oocyte and Cumulus Cells. International Journal of Fertility and Sterility, 12(4):335-338.
22. Sirotkin A.V., Grossmann R. 2008. Effects of ghrelin and its analogues on chicken ovarian granulosa cells. Domestic Animal Endocrinology, 34(2):125-134.
23. Sirotkin A.V., Grossmann R., María-Peon M.T., Roa J., Tena-Sempere M., Klein S. 2006. Novel expression and functional role of ghrelin in chicken ovary. Molecular and Cellular Endocrinology, 257-258:15-25.
24. Stengel A., Taché Y. 2012. Ghrelin - a pleiotropic hormone secreted from endocrine x/a-like cells of the stomach. Frontiers in Neuroscience, 6(24):1-16.
25. Ueberberg B., Unger N., Saeger W., Mann K., Petersenn S. 2009. Expression of ghrelin and its receptor in human tissues. Hormone and Metabolic Research, 41(11):814-821.
26. Wang D., Yang Y., Song Y., Fu S., He X., Wang B, Cao G. 2022. The effect of ghrelin on the maturation of sheep oocytes and early embryonic development in vitro. Animals, 12(9):1158-1173.
27. Yang J., Brown M.S., Liang G., Grishin N., Goldstein J.L. 2008. Identification of the acyltransferase that octanoylates ghrelin, an appetite-stimulating peptide hormone. Cell, 132(3):387-396.
28. Zhu X., Cao Y., Voogd K., Steiner D.F. 2006. On the processing of proghrelin to ghrelin. Journal of Biological Chemistry, 281(50):38867-38870.