Published date: Feb 18 2022

Journal Title: International Journal of Reproductive BioMedicine

Issue title: International Journal of Reproductive BioMedicine (IJRM): Volume 20, Issue No. 1

Pages: 47-58

DOI: 10.18502/ijrm.v20i1.10408

Behrooz NiknafsImmunology Research Center, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran.

Naser ShokrzadehInfertility and Reproductive Health Research Center, Health Research Institute, Babol University of Medical Sciences, Babol, Iran.

Mohammad Reza AlivandDepartment of Genetics, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran.

Mohammad Bakhtiar Hesam Shariatib.hesamshariati@gmail.comDepartment of Anatomical Sciences and Histology, School of Medicine, Kurdistan University of Medical Sciences, Sanandaj, Iran.

Background: The role of glucocorticoids in implantation has been demonstrated.

Objective: This study aimed to evaluate the effect of dexamethasone on endometrial receptivity.

Materials and Methods: In this experimental study, 40 BALB/c female mice aged eight wk old weighing approximately 25.0 ± 1.4 gr were used. The mice were divided into four groups (n = 10/each) of control, dexamethasone (100 μg/kg, intraperitoneal injection), mammalian target of rapamycin (mTOR) inhibitor (PP242) (30 mg/kg, intraperitoneal injection), and dexamethasone and PP242. The endometrial epithelium of the mouse was separated to measure messenger RNA expression of heart and neural crest derivativesexpressed protein 2 (HAND2), Msh homeobox 1 (Msx-1), heparin binding epidermal growth factor (HB-EGF), microRNA (miRNA) Let-7a, miRNA-145 and miRNA-451, using real-time polymerase chain reaction. Also, protein expression of mammalian mTOR and eukaryotic translation initiation factor 4E-binding protein1 (4E-BP1) was measured using western blot.

Results: The results revealed that the expression of Msx-1, HAND2, HB-EGF, miRNA-451, and miRNA-Let-7a was significantly decreased in the endometrium in the dexamethasone group compared to the control, while the expression of miRNA-145 in the endometrium was up-regulated. Additionally, the administration of PP242, known as an inhibitor of mTOR, was associated with significantly reduced expression of Msx-1, HAND2, HB-EGF, miRNA-451, and miRNA-Let-7a, while PP242 induced messenger RNA expression of miRNA-145.

Conclusion: It appears that dexamethasone can diminish uterine receptivity during the implantation period, at least to some extent, through the alteration of particular genes that impact endometrial receptivity. Furthermore, the mTOR pathway seemingly showed an essential role in endometrial receptivity.

Key words: Endometrial receptivity, Dexamethasone, HB-EGF, MSX-1, mTOR, Mouse.

[1] Kao LC, Germeyer A, Tulac S, Lobo S, Yang JP, Taylor RN, et al. Expression profiling of endometrium from women with endometriosis reveals candidate genes for disease-based implantation failure and infertility. Endocrinology 2003; 144: 2870–2881.

[2] Bastu E, Mutlu MF, Yasa C, Dural O, Aytan AN, Celik C, et al. Role of mucin 1 and glycodelin A in recurrent implantation failure. Fertil Steril 2015; 103: 1059– 1064.

[3] Salleh N, Giribabu N. Leukemia inhibitory factor: Roles in embryo implantation and in nonhormonal contraception. Sci World J 2014; 2014: 201514.

[4] Alappat S, Zhang ZY, Chen YP. Msx homeobox gene family and craniofacial development. Cell Res 2003; 13: 429–442.

[5] Nallasamy Sh, Li Q, Bagchi MK, Bagchi IC. Msx homeobox genes critically regulate embryo implantation by controlling paracrine signaling between uterine stroma and epithelium. PLoS Genet 2012; 8: e1002500.

[6] Li Q, Kannan A, DeMayo FJ, Lydon JP, Cooke PS, Yamagishi H, et al. The antiproliferative action of progesterone in uterine epithelium is mediated by Hand2. Science 2011; 331: 912–916.

[7] Cha J, Sun X, Dey SK. Mechanisms of implantation: Strategies for successful pregnancy. Nat Med 2012; 18: 1754–1767.

[8] Chen Ch, Zhao Y, Yu Y, Li R, Qiao J. MiR- 125b regulates endometrial receptivity by targeting MMP26 in women undergoing IVF-ET with elevated progesterone on HCG priming day. Sci Rep 2016; 6: 25302.

[9] Kang YJ, Lees M, Matthews LC, Kimber SJ, Forbes K, Aplin JD. miR-145 suppresses embryo–epithelial juxtacrine communication at implantation by modulating maternal IGF1R. J Cell Sci 2015; 128: 804–814.

[10] Hull ML, Nisenblat V. Tissue and circulating microRNA influence reproductive function in endometrial disease. Reprod Biomed Online 2013; 27: 515–529.

[11] Cheong AWY, Pang RTK, Liu WM, Kottawatta KSA, Lee KF, Yeung WSB. MicroRNA Let-7a and dicer are important in the activation and implantation of delayed implanting mouse embryos. Hum Reprod 2014; 29: 750–762.

[12] Hoang B, Benavides A, Shi Y, Yang Y, Frost P, Gera J, et al. The PP242 mammalian target of rapamycin (mTOR) inhibitor activates extracellular signalregulated kinase (ERK) in multiple myeloma cells via a target of rapamycin complex 1 (TORC1)/eukaryotic translation initiation factor 4E (eIF-4E)/RAF pathway and activation is a mechanism of resistance. J Biol Chem 2012; 287: 21796–21805.

[13] Niknafs B, Hesam Shariati MB, Shokrzadeh N. miR223-3p, HAND2, and LIF expression regulated by calcitonin in the ERK1/2-mTOR pathway during the implantation window in the endometrium of mice. Am J Reprod Immunol 2020; 85: e13333.

[14] Lang F, Pearce D. Regulation of the epithelial Na+ channel by the mTORC2/SGK1 pathway. Nephrol Dial Transplant 2016; 31: 200–205.

[15] Vemulapalli S, Mita A, Alvarado Y, Sankhala K, Mita M. The emerging role of mammalian target of rapamycin inhibitors in the treatment of sarcomas. Target Oncol 2011; 6: 29–39.

[16] Shariati MBH, Niknafs B, Seghinsara AM, Shokrzadeh N, Alivand MR. Administration of dexamethasone disrupts endometrial receptivity by alteration of expression of miRNA 223, 200a, LIF, Muc1, SGK1, and ENaC via the ERK1/2-mTOR pathway. J Cell Physiol 2019; 234: 19629–19639.

[17] Whitten WK. The effect of removal of the olfactory bulbs on the gonads of mice. J Endocrinol 1956; 14: 160–163.

[18] Whirledge ShD, Oakley RH, Myers PH, Lydon JP, DeMayo F, Cidlowski JA. Uterine glucocorticoid receptors are critical for fertility in mice through control of embryo implantation and decidualization. Proc Natl Acad Sci 2015; 112: 15166–15171.

[19] Gleason CE, Frindt G, Cheng ChJ, Ng M, Kidwai A, Rashmi P, et al. mTORC2 regulates renal tubule sodium uptake by promoting ENaC activity. J Clin Invest 2015; 125: 117–128.

[20] Shokrzadeh N, Alivand MR, Abedelahi A, Hessam Shariati MB, Niknafs B. Upregulation of HB-EGF, Msx. 1, and miRNA Let-7a by administration of calcitonin through mTOR and ERK1/2 pathways during a window of implantation in mice. Mol Reprod Dev 2018; 85: 790–801.

[21] Raab G, Klagsbrun M. Heparin-binding EGF-like growth factor. Biochim Biophys Acta 1997; 1333: F179–F199.

[22] Daikoku T, Song H, Guo Y, Riesewijk A, Mosselman S, Das SK, et al. Uterine Msx-1 and Wnt4 signaling becomes aberrant in mice with the loss of leukemia inhibitory factor or Hoxa-10: Evidence for a novel cytokine-homeobox-Wnt signaling in implantation. Mol Endocrinol 2004; 18: 1238–1250.

[23] Daikoku T, Cha J, Sun X, Tranguch S, Xie H, Fujita T, et al. Conditional deletion of Msx homeobox genes in the uterus inhibits blastocyst implantation by altering uterine receptivity. Dev Cell 2011; 21: 1014–1025.

[24] Huyen DV, Bany BM. Evidence for a conserved function of heart-and neural crest derivativesexpressed transcript 2 (Hand2) in mouse and human decidualization. Reproduction 2011; 142: 353–368.

[25] Hesam Shariati MB, Majdi Seghinsara A, Shokrzadeh N, Niknafs B. The effect of fludrocortisone on the uterine receptivity partially mediated by ERK1/2-mTOR pathway. J Cell Physiol 2019; 234: 20098–20110.

[26] Inyawilert W, Fu TY, Lin CT, Tang PC. Let-7-mediated suppression of mucin 1 expression in the mouse uterus during embryo implantation. J Reprod Dev 2015; 61: 138–144.

[27] Friedman RC, Farh KKH, Burge CB, Bartel DP. Most mammalian mRNAs are conserved targets of microRNAs. Genome Res 2009; 19: 92–105.

[28] Gur Ch, Diav-Citrin O, Shechtman S, Arnon J, Ornoy A. Pregnancy outcome after first trimester exposure to corticosteroids: A prospective controlled study. Reprod Toxicol 2004; 18: 93–101.

[29] Li R, Qiao J, Wang L, Li L, Zhen X, Liu P, et al. MicroRNA array and microarray evaluation of endometrial receptivity in patients with high serum progesterone levels on the day of hCG administration. Reprod Biol Endocrinol 2011; 9: 29.

[30] Nothnick WB, Healy C, Hong X. Steroidal regulation of uterine miRNAs is associated with modulation of the miRNA biogenesis components Exportin-5 and Dicer1. Endocrine 2010; 37: 265–273.

[31] Li Z, Jia J, Gou J, Zhao X, Yi T. MicroRNA-451 plays a role in murine embryo implantation through targeting Ankrd46, as implicated by a microarraybased analysis. Fertil Steril 2015; 103: 834–844.

[32] Revel A, Achache H, Stevens J, Smith Y, Reich R. MicroRNAs are associated with human embryo implantation defects. Hum Reprod 2011; 26: 2830– 2840.

[33] Murakami M, Ichisaka T, Maeda M, Oshiro N, Hara K, Edenhofer F, et al. mTOR is essential for growth and proliferation in early mouse embryos and embryonic stem cells. Mol Cell Biol 2004; 24: 6710–6718.

[34] Wang Y, Zhu L, Kuokkanen S, Pollard JW. Activation of protein synthesis in mouse uterine epithelial cells by estradiol-17ß is mediated by a PKC-ERK1/2-mTOR signaling pathway. Proc Natl Acad Sci 2015; 112: E1382–1391.