The effect of progesterone treatment after ovarian induction on endometrial VEGF gene expression and its receptors in mice at pre-implatation time

Document Type : Original Article


1 Deptartment of Anatomical Sciences, Lorestan University of Medical Sciences, Khorramabad, Iran

2 Razi Herbal Medicines Research Center, Lorestan University of Medical Sciences, Khorramabad, Iran


Objective(s): Progestrone is a prequisite for pre-implantation angiogenesis and induce decidual angiogenesis. It is unknown the effect of progestrone administration on the endometrium of hyperstimulated mice at pre-implantation time.
Material and Methods: Adult female NMRI mice were divided in three groups [control group, ovarian stimulated group and progestrone treated mice after ovarian stimulation]. Uterine horn samples removed at pre-implantation time in each group. Motic image Plus 2 software was used to assess the quantitative vascular parameters of endometrium. Gene expression was determined for vascular endothelial growth factor (VEGF), FMS-like tyrosine kinase (FLT) and Kinase insert domain protein receptor (FLK)genes using the real time PCR method. Data analysis was done with LinReg PCR and Rest-RG software.
Results: Comparison between progestrone treated mice after ovarian stimulation with control group showed that increase in rate of VEGF gene expression [0.775] and decrease in rate of FLK [6.072] and FLT [1.711] gene expression. Analysis of the data on quantitative vascular parameters were indicated remarkable increase in quantitative vascular parameters of progestrone treated mice compare to control group.
Conclusion: Biological effect of progestrone on the vascular changes after ovarian stimulation resulted in an increase in VEGF receptors experession, it seems that induced angiogenesis by progesterone could result in better condition for implantation.


1. Okada H, Tsuzuki T, Shindoh H, Nishigaki A, Yasuda K, Kanzaki H. Regulation of decidualization and angiogenesis in the human endometrium: mini review. J Obstet Gynaecol Res 2014;40:1180-1187.
2. Manolea MM, Dijmărescu AL, Popescu FC, Novac MB, DiŢescu D. Evaluation of the implantation site morphology in spontaneous abortion. Rom J Morphol Embryol 2015;56:125-131.
3. Shifren JL, Tseng JF, Zaloudek CJ, Ryan IP, Meng YG, Ferrara N, Jaffe RB, Taylor RN. Ovarian steroid regulation of vascular endothelialgrowth factor in the human endometrium: implications for angiogenesis during the menstrual cycle and in the pathogenesis of endometriosis. J Clin Endocrinol Metab 1996; 81: 3112–3118.
4. Greb RR, Heikinheimo O, Williams RF, Hodgen GD, Goodman AL. Vascular endothelial growth factor in primate endometrium is regulated by oestrogen-receptor and progesterone-receptor ligands in vivo. Hum Reprod 1997; 12: 1280 –1292.
5. Tan W, Chen L, Guo L, Ou X1, Xie D, Quan S. Relationship between macrophages in mouse uteri and angiogenesis in endometrium during the peri-implantation period. Theriogenology 2014;82:1021-1027.
6. Chakraborty I, Das SK, Dey S. Differential expression of vascular endothelial growth factor and its receptor mRNAs in the mouse uterus around the time of implantation. J Endocrinol 1995; 147: 339–352.
7. Plaks V, Birnberg T, Berkutzki T, Sela S, BenYashar A, Kalchenko V, Mor G, Keshet E, Dekel N, Neeman M, Jung S. Uterine DCs are crucial for decidua formation during embryo implantation in mice. J Clin Invest 2008; 118: 3954–3965.
8. Ferrara N and Davis-Smyth T. The biology of vascular endothelialgrowth factor. Endocrinol Rev 1997; 18: 4–25.
9. Paulson RJ, Sauer MV, Lobo RA. Embryo implantation after human in vitro fertilization:importance of endometrial receptivity. Fertil Steril 1990; 53: 870–874.
10. Horcajadas JA, Riesewijk A, Polman J, Van Os R, Pellicer A, Mosselman S, Simon C. Effect of controlled ovarian hyperstimulation in IVF on endometrial gene expression profiles. Mol Hum Reprod 2005; 11: 195–205.
11. Gomez R, Simon C, Remohi J, Pellicer A. Vascular endothelial growthfactor receptor-2 activation induces vascular permeability in hyperstimulated rats, and this effect is prevented by receptor blockade. Endocrinol 2002; 143: 4339–4348.
12. Filicori M, Fazleabas AT, Huhtaniemi I, Licht P, RaoCh V, Tesarik J, Zygmunt M. Novel concepts of human chorionic gonadotropin: reproductive system interactions and potential in the management of infertility. Fertil Steril 2005; 84: 275–284.
13. Kitajima Y, Endo T, Manase K, Nishikawa A, Shibuya M, Kudo R. Gonadotropin-releasing hormone agonist administration reduced vascular endo.thelial growth factor (VEGF), VEGF receptors, and vascular permeability of the ovaries of hyperstimulated rats. Fertil Steril 2004; 81: 842-849.
14. Ozawa H. Steroid hormones, the irreceptors and neuroendocrine system. J Nip- pon Med Sch 2005; 72:316–325.
15. Mendelson CR. Mini review: fetal-maternal hormonal signaling in pregnancy and labor. Mol Endocrinol 2009; 23: 947–954.
16. Ramatha lCY, Bagchi C, Taylor RN, Bagchi MK. Endometrial decidualization: of mice and men. Semin Reprod Med 2010; 28: 17–26.
17. Garcia E, Bouchard P, De Brux J, Berdah J, Frydman R, Schaison G, Milgrom E, et al. Use of immunocytochemistry of progesterone and estrogen receptors for endometrial dating. J Clin Endocrinol Metab 1998; 67: 80–87.
18. Wen L, Chen LH, Li HY, Chang SP, Liao CY, Tsui KH, Sung YJ, Chao KC. Roles of Estrogen and Progesterone in Endometrial Hemodynamics and Vascular Endothelial Growth Factor Production. J Chin Med. Assoc 2009; 72:188–193.
19. Ancelin M, Buteau-Lozano H, Meduri G, Osborne-Pellegrin M, Sordello S, Plouet J & Perrot-Applanat M. A dynamic shift ofVEGF isoforms with a transient and selective progesterone-induced expression of VEGF189 regulates angiogenesis and vascular permeability in human uterus. PNAS 2002; 99: 6023–6028.
20. Salehnia M, Arianmanesh M. Different pattern of pinopodes expression in stimulated mouse endometrium. Exp Anim 2005; 54:349-352.
21. Mönckedieck V, Sannecke C, Husen B, Kumbartski M, Kimmig R, Tötsch M, et al. Progestin inhibit expression of MMPs and of angiogenic factors in the human endometrial lesions in a mouse model. Mol Hum Reprod 2009; 15:633-643.
22. Cha J, Sun X, Dey SK. Mechanisms of implantation: strategies for successful pregnancy 2012; Nat Med. 6:1754–1767.
23. Douglas NC, Tang H, Gomez R, Pytowski B, Hicklin DJ, Sauer CM, Kitajewski J, Sauer  MV, Zimmermann RC. Vascular endothelial growth factor receptor 2 [VEGFR-2] functions to promote uterine decidual angiogenesis during early pregnancy in the mouse. Endocrinol 2009; 6: 3845–3854.
24. Meduri G, Bausero P, Perrot-Applanat M. Expression of vascular endothelial growth factor receptors in the human endometrium: modulation during the menstrual cycle. Biol Reprod 2000; 62: 439–447.
25. Okada H,Tsutsumi A,Imai M,Nakajima T,Yasuda K, Kanzaki H. Estrogen and selective estrogen receptor modulators regulate vascular endothelial growth factor and soluble vascular endothelial growth factor receptor in human endometrial stromal cells. Fertil Steril 2010; 93: 2680-2686.
26. Yue TL, Wang X, Louden CS, Gupta S, Pillarisetti K, Gu JL, Hart TK, Lysko PG, Feuerstein GZ. 2-Methoxyestradiol, an endogenous estrogen metabolite, induces apoptosis in endothelial cells and inhibits angiogenesis: possible role for stress-activated protein kinase signaling pathway and Fas expression. Mol Pharmacol 1997; 51: 951–962.
27. Guenther S, Vrekoussis T, Heublein S, Bayer B, Anz D, Knabl J, et al. Decidual macrophages Are significantly increased in spontaneous miscarriages and over-express FasL: a potential role for macrophages in trophoblast apoptosis. Int J Mol Sci 2012; 6: 9069–9080.
28. Tayade C, Hilchie D, He H, Fang Y, Moons L, Carmeliet P, Foster RA, Croy BA. Genetic deletion of placenta growth factor in mice alters uterine NK cells. J. Immunol 2007; 178:4267–4275.
29. Ashkar AA, Croy BA. Functions of uterine natural killer cells are mediated by interferon gamma production during murine pregnancy. Semin Immunol 2001; 13:235–241.