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Heparin prevents oxidative stress-induced apoptosis in human decidualized endometrial stromal cells

  • Shunsuke Tamaru
  • Takeshi KajiharaEmail author
  • Yumi Mizuno
  • Natsuko Takano
  • Hideno Tochigi
  • Tomomi Sato
  • Osamu Ishihara
Original Paper
  • 8 Downloads

Abstract

Clinical trials have shown that administering heparin during the luteal phase has beneficial effects on implantation and live birth rates. Heparin exerts direct effects on decidual human endometrial stromal cells (HESCs), which are independent of its anticoagulant effect. However, the accurate effects of heparin on the decidualization process remain unidentified. Here, we demonstrate that HESCs become dramatically resistant to oxidative stress upon decidualization, and we hypothesize a possible direct action of heparin on the decidualization of HESCs, which would lead to improved implantation. To test this hypothesis, we established primary HESC cultures and propagated them, and then we decidualized confluent cultures with 8-bromo-cAMP, with medroxyprogesterone acetate, and with or without heparin. We treated the cells with hydrogen peroxide (H2O2) as a source of reactive oxygen species (ROS). Adding heparin to decidualized HESCs induced prolactin secretion. Decidualized HESCs treated with heparin were prevented from undergoing apoptosis induced by oxidative stress. Heparin induced nuclear accumulation of the forkhead transcription factor FOXO1 and expression of its downstream target, the ROS scavenger superoxide dismutase 2. These results demonstrate that heparin-treated decidualized HESCs acquired further resistance to oxidative stress, suggesting that heparin may improve the implantation environment.

Keywords

Endometrial decidualization Heparin FOXO1 SOD2 Oxidative stress 

Notes

Acknowledgements

We would like to thank Ms. S. Mitsui for her assistance with experiments. This work was supported by JSPS KAKENHI (Grant numbers 17K11250 and 16K20204).

Compliance with ethical standards

Conflict of interest

The authors declare having no conflicts of interest.

References

  1. 1.
    Rai R, Cohen H, Dave M, Regan L (1997) Randomized controlled trial of aspirin and aspirin plus heparin in pregnant women with recurrent miscarriage associated with phospholipid antibodies (or antiphospholipid antibodies). BMJ 314:253–257CrossRefGoogle Scholar
  2. 2.
    Rai R, Regan L (2002) Antiphospholipid syndrome in pregnancy: a randomized, controlled trial of treatment. Obstet Gynecol 100:1354CrossRefGoogle Scholar
  3. 3.
    Sebire NJ, Backos M, El Gaddal G, Goldin RD, Regan L (2003) Placental pathology, antiphospholipid antibodies, and pregnancy outcome in recurrent miscarriage patients. Obstet Gynecol 101:258–263Google Scholar
  4. 4.
    Urman B, Ata B, Yakin K, Alatas C, Aksoy S, Mercan R, Balaban B (2009) Luteal phase empirical low molecular weight heparin administration in patients with failed ICSI embryo transfer cycles: a randomized open-labeled pilot trial. Hum Reprod 24:1640–1647CrossRefGoogle Scholar
  5. 5.
    Qublan H, Amarin Z, Dabbas M, Farraj AE, Beni-Merei Z, Al-Akash H, Bdoor AN, Nawasreh M, Malkawi S, Diab F, Al-Ahmad N, Balawneh M, Abu-Salim A (2008) Low-molecular-weight heparin in the treatment of recurrent IVF-ET failure and thrombophilia: a prospective randomized placebo-controlled trial. Hum Fertil (Camb) 11:246–253CrossRefGoogle Scholar
  6. 6.
    Hills FA, Abrahams VM, Gonzalez-Timon B, Franvis J, Clock B, Hinkson L, Rai R, Mor Regan L, Sullivan M, Lam EW, Brosens JJ (2006) Heparin prevents programmed cell death in human trophoblast. Mol Hum Reprod 12:237–243CrossRefGoogle Scholar
  7. 7.
    Fluhr H, Spratte J, Ehrardt J, Steinmuller F, Licht P, Zygmunt M (2010) Heparin and low-molecular-weight heparins modulate the decidualization of human endometrial stromal cells. Fertil Steril 93:2581–2587CrossRefGoogle Scholar
  8. 8.
    Kajihara T, Jones M, Fusi L, Takano M, Feroze-Zaidi F, Pirianov G, Mehmet H, Ishihara O, Higham JM, Lam EW, Brosens JJ (2006) Differetial expression of FOXO1 and FOXO3a confers resistance to oxidative cell death upon endometrial decidualization. Mol Endocrinol 20:2444–2455CrossRefGoogle Scholar
  9. 9.
    Kajihara T, Uchino S, Suzuki M, Itakura A, Brosens JJ, Ishihara O (2011) Human chorionic gonadotropin confers resistance to oxidative stress-induced apoptosis in decidualizing human endometrial stromal cells. Fertil Steril 95:1302–1307CrossRefGoogle Scholar
  10. 10.
    Labied S, Kajihara T, Madureira PA, Fusi L, Jones MC, Higham JM, Varshochi R, Francis JM, Zoumpoulidou G, Essafi A, Fernandez de Mattos S. Lam EW, Brosens JJ (2006) Pregestins regulate the expression and activity of the forkhead transcription factor FOXO1 in differentiating human endometrium. Mol Endocrinol 20:35–44CrossRefGoogle Scholar
  11. 11.
    Brosens JJ, Hayashi N, White JO (1999) Progesterone receptor regulates decidual prolactin expression in differentiating human endometrial stromal cell. Endocrinology 140:4809–4820CrossRefGoogle Scholar
  12. 12.
    Christian M, Zhang X, Schneider-Merck T, Unterman TG, Gellersen B, White JO, Brosens JJ (2002) Cyclic AMP-induced forkhead transcription factor, FKHR, cooperates with CCAAT/enhancer-binding protein b in differentiating human endometrial stromal cells. J Biol Chem 277:20825–20832CrossRefGoogle Scholar
  13. 13.
    Tochigi H, Kajihara T, Mizuno Y, Mizuno Y, Tamaru S, Kamei Y, Okazaki Y, Brosens JJ, Ishihara O (2017) Loss of miR-542-3p enhances IGFBP-1 expression in decidualizing human endometrial stromal cells. Sci Rep 2017:40001CrossRefGoogle Scholar
  14. 14.
    Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(−delta delta C(T)) method. Methods 25:402–408CrossRefGoogle Scholar
  15. 15.
    Gellersen B, Kempf R, Telgmann R, DiMattia GE (1994) Nonpituitary human prolactin gene transcription is independent of Pit-1 and differentially controlled in lyphocytes and in endometrial stroma. Mol Endocrinol 8:356–373Google Scholar
  16. 16.
    Gellersen B, Brosens J (2003) Cyclic AMP and progesterone receptor cross-talk in human endometrium: a decidualizing affair. J Endocrinol 178:357–372CrossRefGoogle Scholar
  17. 17.
    Kajihara T, Brosens JJ, Ishihara O (2013) The role of FOXO1 in the decidual transformation of the endometrium and early pregnancy. Med Mol Morphol 46:61–68CrossRefGoogle Scholar
  18. 18.
    Steptoe PC, Edwards RG (1978) Birth after the reimplantation of a human embryo. Lancet 2(8085):366CrossRefGoogle Scholar
  19. 19.
    Donaghay M, Lessey BA (2007) Uterine receptivity: alterations associated with benign gynecological disease. Semin Reprod Med 25:461–475CrossRefGoogle Scholar
  20. 20.
    Gellersen B, Brosens IA, Brosens JJ (2007) Decidualization of the human endometrium: mechanisms, functions, and clinical perspectives. Semin Reprod Med 25:445–453CrossRefGoogle Scholar
  21. 21.
    Wilcox AJ, Baird DD, Weinberg CR (1999) Time of implantation of the conceptus and loss of pregnancy. N Engl J Med 340:1796–1799CrossRefGoogle Scholar
  22. 22.
    Ricci G, Giolo E, Simeone R (2010) Heparin’s `potential to improve pregnancy rates and outcomes’ is not evidence-based. Hum Reprod Update 16:225–227CrossRefGoogle Scholar
  23. 23.
    Cloke B, Huhtinen K, Fusi L, Kajihara T, Yliheikkila M, Ho KK, Teklenburg G, Lavery S, Jones MC, Trew G, Kim JJ, Lam EW, Cartwright JE, Poutanen M, Brosens JJ (2008) The androgen and progesterone receptors regulate distinct gene networks and cellular functions in decidualizing endometrium. Endocrinolgy 149:4462–4474CrossRefGoogle Scholar
  24. 24.
    Lynch VJ, Tanzer A, Wang Y, Leung FC, Gellersen B, Emera D, Wagner GP (2008) Adaptive changes in the transcription factor HoxA–11 are essential for the evolution of pregnancy in mammals. Proc Natl Acad Sci USA 105:14928–14933CrossRefGoogle Scholar
  25. 25.
    Fluhr H, Spratte J, Heidrich S, Ehrhardt J, Steinmüller F, Zygmunt M (2011) Heparin inhibits interferon-γ signaling in human endometrial stromal cells by interference with the cellular binding of interferon-γ. Fertil Steril 95:1272–1277CrossRefGoogle Scholar
  26. 26.
    Salker M, Teklenburg G, Molokhia M, Lavery S, Trew G, Aojanepong T, Mardon H, Lokugamage AU, Rai R, Landles C, Roelen BA, Quenby S, Kuijk EW, Kavelaars A, Heijnen CJ, Regan L, Macklon NS, Brosens JJ (2010) Natural selection of human embryos: impaired decidualization of endometrium disables embryo-maternal interactions and causes recurrent pregnancy loss. PLoS One 5:e10287CrossRefGoogle Scholar
  27. 27.
    Garzia E, Borgato S, Cozzi V, Doi P, Bulfamante G, Persani L, Cetin I (2004) Lack of expression of endometrial prolactin in early implantation failure: a pilot study. Hum Reprod 19:1911–1916CrossRefGoogle Scholar
  28. 28.
    Jauniaux E, Watson AL, Hempstock J, Bao YP, Skepper JN, Burton GJ (2000) Onset of maternal arterial blood flow and placental oxidative stress. A possible factor in human early pregnancy failure. Am J Pathol 157:2111–2122CrossRefGoogle Scholar
  29. 29.
    Jauniaux E, Poston L, Burton GJ (2006) Placental-related diseases of pregnancy: involvement of oxidative stress and implications in human evolution. Hum Reprod Update 12:747–755CrossRefGoogle Scholar
  30. 30.
    Kops GJ, de Ruiter ND, De Vries-Smits AM, Powell DR, Bos JL, Burgering BM (1999) Direct control of the forkhead transcription factor AFX by protein kinase B. Nature 398:630–634CrossRefGoogle Scholar
  31. 31.
    Brunet A, Bonni A, Zigmond MJ, Lin MZ, Juo P, Hu LS, Anderson MJ, Arden KC, Blenis J, Greenberg ME (1999) Akt promotes cell survival by phosphorylating and inhibiting a Forkhead transcription factor. Cell 96:857–868CrossRefGoogle Scholar
  32. 32.
    Dijkers PF, Medema RH, Pals C, Banerji L, Thomas NS, Lam EW, Burgering BM, Raaijmakers JA, Lammers JW, Koenderman L, Coffer PJ (2000) Forkhead transcription factor FKHR-1 modulates cytokine-dependent transcriptional regulation of p27KIP1. Mol Cell Biol 20:9138–9148CrossRefGoogle Scholar
  33. 33.
    Sunters A, Fernandez de Mottos S, Stahl M, Brosens JJ, Zoumpoulidou G, Saunders CA, Coffer PJ, Medema RH, Coombes RC, Lam EW (2003) FoxO3a transcriptional regulation of Bim controls apoptosis in paclitaxel-treated breast cancer cell lines. J Biol Chem 278:49795–49805CrossRefGoogle Scholar
  34. 34.
    Modur V, Nagarajan R, Evers BM, Milbrandt J (2002) FOXO proteins regulate tumor necrosis factor-related apoptosis inducing ligand expression. Implications for PTEN mutation in prostate cancer. J Biol Chem 277:47928–47937CrossRefGoogle Scholar
  35. 35.
    Suhara T, Kim HS, Kirshenbaum LA, Walsh K (2002) Suppression of Akt signaling induces Fas ligand expression: involvement of caspase and Jun kinase activation in Akt-mediated Fas ligand regulation. Mol Cell Biol 22:680–691CrossRefGoogle Scholar
  36. 36.
    Dijkers PF, Medema RH, Lammers JW, Koenderman L, Coffer PJ (2000) Expression of the pro-apoptotic Bcl-2 family member Bim is regulated by the forkhead transcription factor FKHR-L1. Curr Biol 10:1201–1204CrossRefGoogle Scholar
  37. 37.
    Medema RH, Kops GJ, Bos JL, Burgering BM (2000) AFX-like Forkhead transcription factors mediate cell-cycle regulation by Ras and PKB through p27kip1. Nature 404:782–787CrossRefGoogle Scholar
  38. 38.
    Nemoto S, Finkel T (2002) Redox regulation of forkhead proteins through a p66shc-dependent signaling pathway. Science 295:2450–2452CrossRefGoogle Scholar
  39. 39.
    Essers MA, Weijzen S, de Vries-Smits AM, Saarloos I, de Ruiter ND, Bos JL, Burgering BM, EMBO J (2004) FOXO transcription factor activation by oxidative stress mediated by the small GTPase Ral and JNK. EMBO J 23:4802–4812CrossRefGoogle Scholar
  40. 40.
    Tran H, Brunet A, Grenier JM, Datta SR, Fornace AJ Jr, DiStefano PS, Chiang LW, Greenberg ME (2002) DNA repair pathway stimulated by the forkhead transcription factor FOXO3a through the Gadd45 protein. Science 296:530–534CrossRefGoogle Scholar
  41. 41.
    Di Simone N, Di Niculo F, Sanguinetti M, Ferrazzani S, D’Alessio MC, Castellani R, Bompiani A, Caruso A (2007) Low-molecular weight heparin induces in vitro trophoblast invasiveness: role matrix metalloproteinases and tissue inhibitors. Placenta 28:298–304CrossRefGoogle Scholar
  42. 42.
    Greer I, Hunt BJ (2005) Low molecular weight heparin in pregnancy: current issues. Br J Haematol 128:593–601CrossRefGoogle Scholar
  43. 43.
    Kutteh WH (1996) Antiphospholipid antibody-associated recurrent pregnancy loss: treatment with heparin and low-dose aspirin is superior to low-dose aspirin alone. Am J Obstet Gynecol 174:1584–1589CrossRefGoogle Scholar
  44. 44.
    Spratte J, Schönborn M, Treder N, Bornkessel F, Zygmunt M, Fluhr H (2015) Heparin modulates chemokines in human endometrial stromal cells by interaction with tumor necrosis factor α and thrombin. Fertil Steril 103:1363–1369CrossRefGoogle Scholar
  45. 45.
    Yamamoto H, Fuyama S, Arai S, Sendo F (1985) Inhibition of mouse natural killer cytotoxicity by heparin. Cell Immunol 96:409–417CrossRefGoogle Scholar
  46. 46.
    Johann S, Zoller C, Haas S, Blumel G, Lipp M, Forster R (1995) Sulfated polysaccharide anticoagulants suppress natural killer cell activity in vitro. Thromb Haemost 74:998–1002CrossRefGoogle Scholar
  47. 47.
    Christopherson KW 2nd, Campbell JJ, Travers JB, Hromas RA (2002) Low-molecular-weight heparins inhibit CCL2l-induced T cell adhesion and migration. J Pharmacol Exp Ther 302:290–295CrossRefGoogle Scholar
  48. 48.
    Manduteanu I, Voinea M, Capraru M, Dragomir E, Simionescu M (2002) A novel attribute of enoxaparin: inhibition of monocyte adhesion to endothelial cells by a mechanism involving cell adhesion molecules. Pharmacology 65:52–53CrossRefGoogle Scholar
  49. 49.
    Wan JG, Mu JS, Zhu HS, Geng JG (2002) N-desulfated non-anticoagulant heparin inhibits leukocyte adhesion and transmigration in vitro and attenuates acute peritonitis and ischemia and reperfusion injury in vitro. Inflamm Res 51:435–443CrossRefGoogle Scholar
  50. 50.
    Fritchley SJ, Kirby JA, Ali S (2000) The antagonism of interferon-gamma (INF-gamma) by heparin: examination of the blockade of class II MHC antigen and heat shock protein-70 expression. Clin Exp Med Biol 120:247–252Google Scholar
  51. 51.
    Folkman J, Shing Y (1992) Control of angiogenesis by heparin and other sulfated polysaccharides. Adv Exp Med Biol 313:355–364CrossRefGoogle Scholar
  52. 52.
    Li Y, Wang HY, Cho CH (1999) Association of heparin with basic fibroblast growth factor, epidermal growth factor, and constitutive nitric oxide synthase on healing of gastric ulcer in rats. J Pharmacol Exp Ther 290:789–796Google Scholar

Copyright information

© The Japanese Society for Clinical Molecular Morphology 2019

Authors and Affiliations

  1. 1.Department of Obstetrics and GynecologySaitama Medical UniversitySaitamaJapan
  2. 2.Department of AnatomySaitama Medical UniversitySaitamaJapan

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