Skip to main content

Decreased fatty acids induced granulosa cell apoptosis in patients with diminished ovarian reserve

Journal of Assisted Reproduction and Genetics volume 39, pages 1105–1114 (2022)Cite this article

Abstract

Purpose

To investigate whether fatty acid changes in granulosa cells (GCs) underly the pathogenic mechanisms of diminished ovarian reserve (DOR).

Methods

GCs were obtained from patients with DOR (n = 70) and normal ovarian reserve (NOR, n = 70). Analysis of fatty acids changes in GCs was then analyzed.

Results

Patients with DOR had significantly lower levels of antral follicle count and anti-Mullerian hormone and higher levels of follicle-stimulating hormone compared with NOR patients (P < 0.001). The good-quality embryo rate was notably decreased in DOR patients (51.99 vs 39.52%, P < 0.05). A total of 15 significantly decreased fatty acids in GCs from patients with DOR. The ATP levels were markedly lower in DOR patients than in NOR patients (39.07 ± 12.89 vs 23.21 ± 13.69%, P < 0.05). Mitochondrial membrane potential decreased in DOR patients (P < 0.01). In GCs from DOR patients, the β-oxidation genes (HADHA and ACSL) and DNA repair genes (PRKDC and RAD50) were significantly downregulated (P < 0.05). The γH2AX foci/nucleus ratio in DOR patients markedly increased relative to that of NOR patients (0.31 ± 0.03 vs 0.87 ± 0.07, P < 0.001). Meanwhile, the apoptosis rate of GCs was significantly higher in DOR patients (6.43 ± 2.11 vs 48.06 ± 6.72%, P < 0.01).

Conclusion

GC apoptosis resulting from the decrease of fatty acids, and associated with reduced ATP production and DNA damage, may contribute to the pathogenic mechanisms responsible for DOR.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

USD 39.95

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Fig. 1
Fig. 2
Fig. 3
Fig. 4

References

  1. Greene AD, Patounakis G, Segars JH. Genetic associations with diminished ovarian reserve: a systematic review of the literature. J Assist Reprod Genet. 2014;31(8):935–46.

    Article  PubMed  PubMed Central  Google Scholar 

  2. Pal L, Bevilacqua K, Santoro NF. Chronic psychosocial stressors are detrimental to ovarian reserve: a study of infertile women. J Psychosom Obstet Gynaecol. 2010;31(3):130–9.

    Article  PubMed  Google Scholar 

  3. Mutlu MF, Erdem A. Evaluation of ovarian reserve in infertile patients. J Turk Ger Gynecol Assoc. 2012;13(3):196–203.

    Article  PubMed  PubMed Central  Google Scholar 

  4. Gilchrist RB, Ritter LJ, Armstrong DT. Oocyte-somatic cell interactions during follicle development in mammals. Anim Reprod Sci. 2004;82–83:431–46.

    Article  PubMed  CAS  Google Scholar 

  5. Dunning KR, Russell DL, Robker RL. Lipids and oocyte developmental competence: the role of fatty acids and β-oxidation. Reproduction. 2014;148(1):R15-27.

    Article  CAS  PubMed  Google Scholar 

  6. Downs SM, Mosey JL, Klinger J. Fatty acid oxidation and meiotic resumption in mouse oocytes. Mol Reprod Dev. 2009;76(9):844–53.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Sturmey RG, O’Toole PJ, Leese HJ. Fluorescence resonance energy transfer analysis of mitochondrial:lipid association in the porcine oocyte. Reproduction. 2006;132(6):829–37.

    Article  CAS  PubMed  Google Scholar 

  8. Dunning KR, et al. Beta-oxidation is essential for mouse oocyte developmental competence and early embryo development. Biol Reprod. 2010;83(6):909–18.

    Article  CAS  PubMed  Google Scholar 

  9. Zhao Z, et al. The mRNA expression signature and prognostic analysis of multiple fatty acid metabolic enzymes in clear cell renal cell carcinoma. J Cancer. 2019;10(26):6599–607.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Bradley J, Swann K. Mitochondria and lipid metabolism in mammalian oocytes and early embryos. Int J Dev Biol. 2019;63(3–4–5):93–103.

    Article  CAS  PubMed  Google Scholar 

  11. Dunning KR, et al. Increased beta-oxidation and improved oocyte developmental competence in response to l-carnitine during ovarian in vitro follicle development in mice. Biol Reprod. 2011;85(3):548–55.

    Article  CAS  PubMed  Google Scholar 

  12. Nagata S, et al. Effect of aging on mitochondria and metabolism of bovine granulosa cells. J Reprod Dev. 2020;66(6):547–54.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Ge H, et al. The importance of mitochondrial metabolic activity and mitochondrial DNA replication during oocyte maturation in vitro on oocyte quality and subsequent embryo developmental competence. Mol Reprod Dev. 2012;79(6):392–401.

    Article  CAS  PubMed  Google Scholar 

  14. Babayev E, Seli E. Oocyte mitochondrial function and reproduction. Curr Opin Obstet Gynecol. 2015;27(3):175–81.

    Article  PubMed  PubMed Central  Google Scholar 

  15. de Barros TT, et al. DNA damage is inversely associated to blood levels of DHA and EPA fatty acids in Brazilian children and adolescents. Food Funct. 2020;11(6):5115–21.

    Article  PubMed  Google Scholar 

  16. Zhang D, et al. Increased DNA damage and repair deficiency in granulosa cells are associated with ovarian aging in rhesus monkey. J Assist Reprod Genet. 2015;32(7):1069–78.

    Article  PubMed  PubMed Central  Google Scholar 

  17. Qian M, et al. Boosting ATM activity alleviates aging and extends lifespan in a mouse model of progeria. Elife. 2018;7.e348360

  18. Lee JH, Paull TT. ATM activation by DNA double-strand breaks through the Mre11-Rad50-Nbs1 complex. Science. 2005;308(5721):551–4.

    Article  CAS  PubMed  Google Scholar 

  19. Goodwin JF, Knudsen KE. Beyond DNA repair: DNA-PK function in cancer. Cancer Discov. 2014;4(10):1126–39.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Roos WP, Kaina B. DNA damage-induced cell death: from specific DNA lesions to the DNA damage response and apoptosis. Cancer Lett. 2013;332(2):237–48.

    Article  CAS  PubMed  Google Scholar 

  21. Fan Y, et al. Apoptosis of mural granulosa cells is increased in women with diminished ovarian reserve. J Assist Reprod Genet. 2019;36(6):1225–35.

    Article  PubMed  PubMed Central  Google Scholar 

  22. Cil AP, et al. Assessment of ovarian reserve and Doppler characteristics in patients with multiple sclerosis using immunomodulating drugs. J Turk Ger Gynecol Assoc. 2009;10(4):213–9.

    PubMed  PubMed Central  Google Scholar 

  23. Testing and interpreting measures of ovarian reserve: a committee opinion. Fertil Steril. 2020;114(6):1151-1157.

  24. Li R, Albertini DF. The road to maturation: somatic cell interaction and self-organization of the mammalian oocyte. Nat Rev Mol Cell Biol. 2013;14(3):141–52.

    Article  CAS  PubMed  Google Scholar 

  25. Huang R, et al. Alterations of polyunsaturated fatty acid metabolism in ovarian tissues of polycystic ovary syndrome rats. J Cell Mol Med. 2018;22(7):3388–96.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Sawyer BT, et al. Targeting fatty acid oxidation to promote anoikis and inhibit ovarian cancer progression. Mol Cancer Res. 2020;18(7):1088–98.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. McKeegan PJ, Sturmey RG. The role of fatty acids in oocyte and early embryo development. Reprod Fertil Dev. 2011;24(1):59–67.

    Article  CAS  PubMed  Google Scholar 

  28. O’Gorman A, et al. Metabolic profiling of human follicular fluid identifies potential biomarkers of oocyte developmental competence. Reproduction. 2013;146(4):389–95.

    Article  CAS  PubMed  Google Scholar 

  29. Dubeibe Marin DF, et al. Importance of lipid metabolism on oocyte maturation and early embryo development: can we apply what we know to buffalo? Anim Reprod Sci. 2019;211:106220.

    Article  CAS  PubMed  Google Scholar 

  30. Dunning KR, et al. Regulation of fatty acid oxidation in mouse cumulus-oocyte complexes during maturation and modulation by PPAR agonists. PLoS One. 2014;9(2):e87327.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  31. Ohata K, et al. Effects of fatty acid supplementation during vitrification and warming on the developmental competence of mouse, bovine and human oocytes and embryos. Reprod Biomed Online. 2021;43(1):14–25.

    Article  CAS  PubMed  Google Scholar 

  32. Wang HW, et al. Activity of long-chain acyl-CoA synthetase is required for maintaining meiotic arrest in Xenopus laevis. Biol Reprod. 2012;87(3):74.

    Article  PubMed  CAS  Google Scholar 

  33. Igarashi H, et al. Aged mouse oocytes fail to readjust intracellular adenosine triphosphates at fertilization. Biol Reprod. 2005;72(5):1256–61.

    Article  CAS  PubMed  Google Scholar 

  34. Sullivan EM, et al. Mechanisms by which dietary fatty acids regulate mitochondrial structure-function in health and disease. Adv Nutr. 2018;9(3):247–62.

    Article  PubMed  PubMed Central  Google Scholar 

  35. Dumesic DA, et al. Cumulus cell mitochondrial resistance to stress in vitro predicts oocyte development during assisted reproduction. J Clin Endocrinol Metab. 2016;101(5):2235–45.

    Article  CAS  PubMed  Google Scholar 

  36. Xu G, et al. Cadmium induces apoptosis of human granulosa cell line KGN via mitochondrial dysfunction-mediated pathways. Ecotoxicol Environ Saf. 2021;220:112341.

    Article  CAS  PubMed  Google Scholar 

  37. Lee KS, et al. Cumulus cells apoptosis as an indicator to predict the quality of oocytes and the outcome of IVF-ET. J Assist Reprod Genet. 2001;18(9):490–8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Banim PJ, et al. Dietary oleic acid is inversely associated with pancreatic cancer - data from food diaries in a cohort study. Pancreatology. 2018;18(6):655–60.

    Article  CAS  PubMed  Google Scholar 

  39. Winship AL, et al. The importance of DNA repair for maintaining oocyte quality in response to anti-cancer treatments, environmental toxins and maternal ageing. Hum Reprod Update. 2018;24(2):119–34.

    Article  CAS  PubMed  Google Scholar 

  40. Stringer JM, et al. The capacity of oocytes for DNA repair. Cell Mol Life Sci. 2018;75(15):2777–92.

    Article  CAS  PubMed  Google Scholar 

  41. Bedoschi GM, Navarro PA, Oktay KH. Novel insights into the pathophysiology of chemotherapy-induced damage to the ovary. Panminerva Med. 2019;61(1):68–75.

    Article  PubMed  Google Scholar 

  42. Glamoclija V, et al. Apoptosis and active caspase-3 expression in human granulosa cells. Fertil Steril. 2005;83(2):426–31.

    Article  CAS  PubMed  Google Scholar 

  43. Stoffel W, et al. Dietary ω3-and ω6-polyunsaturated fatty acids reconstitute fertility of juvenile and adult Fads2-deficient mice. Mol Metab. 2020;36:100974.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Zarezadeh R, et al. Fatty acids of follicular fluid phospholipids and triglycerides display distinct association with IVF outcomes. Reprod Biomed Online. 2021;42(2):301–9.

    Article  CAS  PubMed  Google Scholar 

  45. Yang Y, et al. Metabolic changes of maternal uterine fluid, uterus, and plasma during the peri-implantation period of early pregnancy in mice. Reprod Sci. 2020;27(2):488–502.

    Article  PubMed  Google Scholar 

  46. Karaşahin T. The effect of oleic and linoleic acid addition to the culture media on bovine embryonic development following vitrification. Pol J Vet Sci. 2019;22(4):661–6.

    PubMed  Google Scholar 

  47. Haggarty P, et al. Fatty acid metabolism in human preimplantation embryos. Hum Reprod. 2006;21(3):766–73.

    Article  CAS  PubMed  Google Scholar 

  48. Hohos NM, et al. Fat-1 transgene is associated with improved reproductive outcomes. Endocrinology. 2018;159(12):3981–92.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  49. Sun X, et al. Research on serum metabolomics of ovariectomized rats and intervention effect of Cuscuta chinensis on metabolic pattern. J Pharm Biomed Anal. 2021;195:113847.

    Article  CAS  PubMed  Google Scholar 

  50. Wu TC, Wang L, Wan YJ. Expression of estrogen receptor gene in mouse oocyte and during embryogenesis. Mol Reprod Dev. 1992;33(4):407–12.

    Article  CAS  PubMed  Google Scholar 

  51. Panagiotopoulos AA, et al. Eicosanoids in prostate cancer. Cancer Metastasis Rev. 2018;37(2–3):237–43.

    Article  CAS  PubMed  Google Scholar 

  52. Khajeh M, et al. Potential role of polyunsaturated fatty acids, with particular regard to the signaling pathways of arachidonic acid and its derivatives in the process of maturation of the oocytes: contemporary review. Biomed Pharmacother. 2017;94:458–67.

    Article  CAS  PubMed  Google Scholar 

  53. Hoyos-Marulanda V, et al. Effects of polyunsaturated fatty acids on the development of pig oocytes in vitro following parthenogenetic activation and on the lipid content of oocytes and embryos. Anim Reprod Sci. 2019;205:150–5.

    Article  CAS  PubMed  Google Scholar 

  54. Jahangirifar M, et al. Dietary fatty acid intakes and the outcomes of assisted reproductive technique in infertile women. J Reprod Infertil. 2021;22(3):173–83.

    PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

This research was supported by the National Natural Science Foundation of China(No.81960278), the Outstanding Youth Funds of Science and Technology Department of Gansu Province (No.20JR5RA371)(No.20JR10RA701), and Fundamental Research Funds for the Central Universities (No.lzujbky-2021-kd38)

Author information

Authors and Affiliations

  1. The First Clinical Medical College of Lanzhou University, Lanzhou, China

    Zhongying Zhao, Qigang Fan, Qinying Zhu, Ruifen He & Yi Li

  2. Department of Obstetrics and Gynecology, Key Laboratory for Gynecologic Oncology Gansu Province, The First Hospital of Lanzhou University, The First Clinical Medical College of Lanzhou University, Lanzhou, China

    Chang Liu, Jing Wang & Xiaolei Liang

Authors
  1. Zhongying Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Qigang Fan
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Qinying Zhu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ruifen He
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Yi Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Chang Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Jing Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Xiaolei Liang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Xiaolei Liang.

Ethics declarations

Ethics approval

The study was approved by the ethics committee of the First Hospital of Lanzhou University (LDYYLL2019-44).

Consent to participate

Available.

Conflict of interest

The authors declare no competing interests.

Additional information

Publisher's note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary file1 (DOCX 245 KB)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Zhao, Z., Fan, Q., Zhu, Q. et al. Decreased fatty acids induced granulosa cell apoptosis in patients with diminished ovarian reserve. J Assist Reprod Genet 39, 1105–1114 (2022). https://doi.org/10.1007/s10815-022-02462-8

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10815-022-02462-8

Keywords

  • Diminished ovarian reserve
  • Fatty acids
  • ATP production
  • DNA damage
  • Granulosa cell apoptosis