Investigation of the endometrial receptivity status in experimental hypothyroid-induced female rats

Document Type : Original Article

Authors

Department of Histology and Embryology, Faculty of Veterinary Medicine, Atatürk University, Erzurum, Turkey

10.22038/ijbms.2022.63143.13948

Abstract

Objective(s): This study aimed to investigate hypothyroidism’s effects on endometrial receptivity, creating an experimental hypothyroidism model in female rats. 
Materials and Methods: To induce hypothyroidism in rats of Hipotiroid-ER and Treatment-ER groups, 0.05% 6-propyl-2-thiouracil was freshly added to their drinking water for 8 weeks and then the endometrial receptivity model was applied and sacrificed on the fifth day. In the Treatment-ER group, after sc-administration of 0.8 µg/100 g L-thyroxine for 10 days, the endometrial receptivity model was applied to the rats and sacrificed on the fifth day. 
Results: In the histopathological evaluation, epithelial degeneration, vacuolization, enlargement of the uterine glands, and morphological disorders were observed in the endometrial layer of the Hypothyroid-ER group. However, these pathologies were significantly alleviated in the Treatment-ER group. Integrin β3, integrin αvβ3, LIF, and HOXA10 immune reaction intensities were high in the Control-ER and Treatment-ER groups, while in the Hypothyroid-ER group, integrin β3, integrin αvβ3, LIF, and HOXA10 immunoreactivity intensities were low. Also, while MUC1 immunoreactivity was high in the Hypothyroid-ER group, it was low in the other groups. In biochemical analysis, a significant increase in the TSH and progesterone levels and a significant decrease in the FT4, E2, FSH, and LH levels in the Hypothyroid-ER group compared with the Control-ER group were observed. Also, all hormone levels were significantly ameliorated in the rats of the Treatment-ER group compared with the Hypothyroid-ER group. 
Conclusion: The results obtained showed that hypothyroidism had a significant effect on endometrial receptivity—the histopathological and biochemical changes caused by hypothyroidism in the experimental rat model were ameliorated with L-thyroxine treatment.

Keywords


1. Çelik Ö. Yardımcı üreme teknikleri: Temel klinik ve embriyolojik uygulamalar. Nobel Kitabevi 2011. 
2.    Speroff L, Fritz MA. Clinical gynecologic endocrinology and infertility. Edition. Philadelphia, Lippincott Williams & Wilkins 2005.
3.    Unuane D, Poppe K. Female infertility: do we forget the thyroid? J Endocrinol Invest 2015; 38: 571-574.
4.    Dunn D, Turner C. Hypothyroidism in women. Nurs womens Health 2016; 20: 93-98.
5.    Boyar Hİ. Kadın infertilitesi ve endokrinolojik hastalıklar. Dicle Tıp Dergisi 2013; 40: 700-703.
6.    Panciera D, Purswell B, Kolster K. Effect of short-term hypothyroidism on reproduction in the bitch. Theriogenology 2007; 68: 316-321.
7.    Armada-Dias L, Carvalho J, Breitenbach M, Franci C, Moura E. Is the infertility in hypothyroidism mainly due to ovarian or pituitary functional changes? Braz J Med Biol Res 2001; 34: 1209-1215.
8.    Lutsyk A, Sogomonian E. Structural, functional, and lectin histochemical characteristics of rat ovaries and endometrium in experimental hyper-and hypothyroidism. Folia Histochem Cytobiol 2012; 50: 331-339.
9.    Saran S, Gupta BS, Philip R, Singh KS, Bende SA, Agroiya P, Agrawal P. Effect of hypothyroidism on female reproductive hormones. Indian J Endocrinol Metab 2016; 20: 108-113.
10.    Inuwa I, Williams M. A morphometric study on the endometrium of rat uterus in hypothyroid and thyroxine treated hypothyroid rats. Ups J Med Sci 2006; 111: 215-226.
11.    Narvekar SA, Gupta N, Shetty N, Kottur A, Srinivas M, Rao KA. Does local endometrial injury in the nontransfer cycle improve the IVF-ET outcome in the subsequent cycle in patients with previous unsuccessful IVF? A randomized controlled pilot study. J Hum Reprod Sci 2010; 3: 15-19.
12.    Wang L, Qiao J, Li R, Zhen X, Liu Z. Role of endometrial blood flow assessment with color   Doppler energy in predicting pregnancy outcome of IVF-ET cycles. Reprod Biol Endocrinol 2010; 8: 122-128.
13.    Achache H, Revel A. Endometrial receptivity markers, the journey to successful embryo implantation. Hum Reprod Update 2006; 12: 731-746.
14.    Cavagna M, Mantese J. Biomarkers of endometrial receptivity—a review. Placenta 2003; 24: S39-S47.
15.    Lim H, Ma L, Ma W-g, Maas RL, Dey SK. Hoxa-10 regulates uterine stromal cell responsiveness to progesterone during implantation and decidualization in the mouse. Mol Endocrinol 1999; 13: 1005-1017.
16.    Stewart CL. Leukaemia inhibitory factor and the regulation of blastocyst implantation.   Endocrinology of Embryo-Endometrium Interactions, Springer 1994; 269-278.
17.    Surveyor GA, Gendler SJ, Pemberton L, Das S, Chakraborty I, Julian J, Pimental R, Wegner C, Dey S, Carson D. Expression and steroid hormonal control of Muc-1 in the mouse uterus. Endocrinology 1995; 136: 3639-3647.
18.    Quaas A, Dokras A. Diagnosis and treatment of unexplained infertility. Rev Obstet Gynecol 2008; 1: 69-76.
19.    Moulakakis KG, Poulakou MV, Dosios T, Dontas I, Sokolis DP, VLACHOS IS, Safioleas MC, Papachristodoulou A, Karayannacos PE, Perrea DN. Hypothyroidism and the aorta. Evidence of increased oxidative DNA damage to the aorta of hypothyroid rats. In Vivo 2008; 22: 603-608.
20.    Yener T, Turkkani Tunc A, Aslan H, Aytan H, Cantug Caliskan A. Determination of oestrous cycle of the rats by direct examination: how reliable? Anat Histol Embryol 2007; 36: 75-77.
21.    Çoban ZD, Güran Ş, Altaylı E, Kayır H, Baykal B. Farede östrus siklusu tayininde hızlı, kolay ve etkin bir yöntem. Gülhane Tıp Derg 2016; 58: 82-87.
22.    Shan L, Zhou Y, Peng S, Wang X, Shan Z, Teng W. Implantation failure in rats with subclinical hypothyroidism is associated with LIF/STAT3 signaling. Endocr Connect 2019; 8: 718-727.
23.    Voipio H-M, Nevalainen T. Improved method for vaginal plug detection in rats. Scand J Lab Anim Sci 1998; 25: 5-9.
24.    Akpak YK, Çekmez Y, Erdoğan Çakır A, Karaca N, Batmaz G, Gülşen S, Tuştaş Haberal E. An animal model of effects of nicotine exposure on endometrial receptivity and embryo implantation in pregnancy. J Matern Fetal Neonatal Med 2017; 30: 2818-2823.
25.    Choudhury KR, Yagle KJ, Swanson PE, Krohn KA, Rajendran JG. A robust automated measure of average antibody staining in immunohistochemistry images. J Histochem Cytochem 2010; 58: 95-107.
26.    Klein M, Vignaud J-M, Hennequin V, Toussaint B, Bresler L, Plenat F, Leclere J, Duprez A, Weryha G. Increased expression of the vascular endothelial growth factor is a pejorative prognosis marker in papillary thyroid carcinoma. J Clin Endocrinol Metab 2001; 86: 656-658.
27.    Dittrich R, Beckmann MW, Oppelt PG, Hoffmann I, Lotz L, Kuwert T, Mueller A. Thyroid hormone receptors and reproduction. J Reprod Immunol 2011; 90: 58-66.
28.    Çakır N. Gebelik ve tiroid hastalıkları. Turk J Endocrinol Metab 2003; 7: 053-057.
29.    Alvarez-Dolado M, Cuadrado A, Navarro-Yubero C, Sonderegger P, Furley AJ, Bernal J, Muñoz A. Regulation of the L1 cell adhesion molecule by thyroid hormone in the developing brain. Mol Cell Neurosci 2000; 16: 499-514.
30.    Legrand J. Effects of thyroid hormones on central nervous system development. Neurobehavioral Teratology 1984: 331-363.
31.    Parija SC, Raviprakash V, Telang AG, Varshney VP, Mishra SK. Influence of hypothyroid state on 45Ca2+ influx and sensitivity of rat uterus to nifedipine and diltiazem. Eur J Pharmacol 2001; 421: 207-213.
32.    Mangge H, Prüller F, Zelzer S, Ainödhofer H, Pailer S, Kieslinger P, Haybaeck J, Obermayer-Pietsch B, Cvirn G, Gruber H-J. Hypothyroidism Exacerbates Thrombophilia in Female Rats Fed with a High Fat Diet. Int J Mol Sci 2015; 16: 15776-15784.
33.    Chang Y-J, Hwu C-M, Yeh C-C, Wang PS, Wang S-W. Effects of subacute hypothyroidism on metabolism and growth-related molecules. Molecules 2014; 19: 11178-11195.
34.    Bentin-Ley U. Relevance of endometrial pinopodes for human blastocyst implantation. Hum Reprod 2000; 15: 67-73.
35.    Bagot CN, Kliman HJ, Taylor HS. Maternal Hoxa10 is required for pinopod formation in the development of mouse uterine receptivity to embryo implantation. Dev Dyn 2001; 222: 538-544.
36.    Afami M. Identify and characterise stem cell population of the human endometrium. Lancaster University 2015.
37.    Kayhan HS. Hipotiroidik Sıçanlarda Arı Sütü Takviyesinin Uterus Üzerindeki Olası Koruyucu Etkisi. Histoloji ve Embriyoloji Anabilim Dalı. Gazi Üniversitesi 2018.
38.    Bowen JA, Hunt JS. The role of integrins in reproduction. Proc Soc Exp Biol Med 2000; 223: 331-343.
39.    Srinivasan KR, Blesson CS, Fatima I, Kitchlu S, Jain SK, Mehrotra PK, Dwivedi A. Expression of αVβ3 integrin in rat endometrial epithelial cells and its functional role during implantation. Gen Comp Endocrinol 2009; 160: 124-133.
40.    Merviel P, Challier J-C, Carbillon L, Foidart J-M, Uzan S. The role of integrins in human embryo implantation. Fetal Diagn Ther 2001; 16: 364-371.
41.    Wu Z, Cai Y, Xia Q, Liu T, Yang H, Wang F, Wang N, Yu Z, Yin C, Wang Q. Hashimoto’s thyroiditis impairs embryo implantation by compromising endometrial morphology and receptivity markers in euthyroid mice. Reprod Biol Endocrinol 2019; 17: 1-13.
42.    Tei C, Maruyama T, Kuji N, Miyazaki T, Mikami M, Yoshimura Y. Reduced expression of αvβ3 integrin in the endometrium of unexplained infertility patients with recurrent IVF-ET failures: improvement by danazol treatment. J Assist Reprod Genet 2003; 20: 13-20.
43.    Taylor HS, Arici A, Olive D, Igarashi P. HOXA10 is expressed in response to sex steroids at the time of implantation in the human endometrium. J Clin Invest 1998; 101: 1379-1384.
44.    Daftary GS, Taylor HS. Molecular markers of implantation: clinical implications. Curr Opin Obstet Gynecol 2001; 13: 269-274.
45.    Cermik D, Selam B, Taylor HS. Regulation of HOXA-10 expression by testosterone in vitro and in the endometrium of patients with polycystic ovary syndrome. J Clin Endocrinol Metab 2003; 88: 238-243.
46.    Matsuzaki S, Canis M, Darcha C, Pouly J-L, Mage G. HOXA-10 expression in the mid-secretory endometrium of infertile patients with either endometriosis, uterine fibromas or unexplained infertility. Hum Reprod 2009; 24: 3180-3187.
47.    Bhatt H, Brunet LJ, Stewart CL. Uterine expression of leukemia inhibitory factor coincides with the onset of blastocyst implantation. Proc Natl Acad Sci U S A 1991; 88: 11408-11412.
48.    Stewart CL, Kaspar P, Brunet LJ, Bhatt H, Gadi I, Köntgen F, Abbondanzo SJ. Blastocyst implantation depends on maternal expression of leukaemia inhibitory factor. Nature 1992; 359: 76-79.
49.    Stewart CL. The role of leukemia inhibitory factor (LIF) and other cytokines in regulating implantation in mammals. Ann NY Acad Sci 1994; 734: 157-165.
50.    Tawfeek MA, Eid MA, Hasan AM, Mostafa M, El-Serogy HA. Assessment of leukemia inhibitory factor and glycoprotein 130 expression in endometrium and uterine flushing: a possible diagnostic tool for impaired fertility. BMC Womens Health 2012; 12: 10-15.
51.    Steck T, Giess R, Suetterlin MW, Bolland M, Wiest S, Poehls UG, Dietl J. Leukaemia inhibitory factor (LIF) gene mutations in women with unexplained infertility and recurrent failure of implantation after IVF ande   embryo transfer. Eur J Obstet Gynecol Reprod Biol 2004; 112: 69-73.
52.    Serafini PC, Silva ID, Smith GD, Motta EL, Rocha AM, Baracat EC. Endometrial claudin-4 and leukemia inhibitory factor are associated with assisted reproduction outcome. Reprod Biol Endocrinol 2009; 7: 30-38.
53.    Hey NA, Graham RA, Seif MW, Aplin JD. The polymorphic epithelial mucin MUC1 in human endometrium is regulated with maximal expression in the implantation phase. J Clin Endocrinol Metab 1994; 78: 337-342.
54.    Tabibzadeh S. Molecular control of the implantation window. Hum Reprod Update 1998; 4: 465-471.
55.    Horne AW, White JO, Lalani E-N, Mobberley MA, Margara RA, Trew GH, Ryder TA. Analysis of epitopes on endometrial epithelium by scanning immunoelectron microscopy. Biochem Biophys Res Commun 2002; 292: 102-108.
56.    Aplin J, Hey N, Li T. MUC1 as a cell surface and secretory component of endometrial epithelium: reduced levels in recurrent miscarriage. Am J Reprod Immunol 1996; 35: 261-266.
57.    Bastu E, Mutlu MF, Yasa C, Dural O, Aytan AN, Celik C, Buyru F, Yeh J. Role of Mucin 1 and Glycodelin A in recurrent implantation failure. Fertil Steril 2015; 103: 1059-1064.e2.
58.    Xu B, Sun X, Li L, Wu L, Zhang A, Feng Y. Pinopodes, leukemia inhibitory factor, integrin-β3, and mucin-1 expression in the peri-implantation endometrium of women with unexplained recurrent pregnancy loss. Fertil Steril 2012; 98: 389-395.
59.    Margarit L, Taylor A, Roberts M, Hopkins L, Davies C, Brenton A, Conlan R, Bunkheila A, Joels L, White J. MUC1 as a discriminator between endometrium from fertile and infertile patients with PCOS and endometriosis. J Clin Endocrinol Metab 2010; 95: 5320-5329.
60.    Horne A, Lalani EN, Margara R, Ryder T, Mobberley M, White J. The expression pattern of MUC1 glycoforms and other biomarkers of endometrial receptivity in fertile and infertile women. Mol Reprod Dev 2005, 72: 216-229.
61.    Hatsuta M, Abe K, Tamura K, Ryuno T, Watanabe G, Taya K, Kogo H. Effects of hypothyroidism on the estrous cycle and reproductive hormones in mature female rat. Eur J Pharmacol 2004; 486: 343-348.
62.    Saleh AAS. Lipid profile and levels of homocysteine and total antioxidant capacity in plasma of rats with experimental thyroid disorders. J Basic App Zool 2015; 72: 173-178.