Microvascular density and vascular endothelial growth factor and osteopontin expression during the implantation window in a controlled ovarian hyperstimulation rat model

Gong,Xin; Tong,Qing; Chen,Zhenzhen; Zhang,Yunna; Xu,Cai; Jin,Zhe

doi:10.3892/etm.2015.2181

Microvascular density and vascular endothelial growth factor and osteopontin expression during the implantation window in a controlled ovarian hyperstimulation rat model

Authors:
- Xin Gong
- Qing Tong
- Zhenzhen Chen
- Yunna Zhang
- Cai Xu
- Zhe Jin
View Affiliations

Affiliations: Reproductive Endocrinology Center, Dongfang Hospital of Beijing University of Chinese Medicine, Beijing 100078, P.R. China, School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 100102, P.R. China
Published online on: January 14, 2015 https://doi.org/10.3892/etm.2015.2181
Pages: 773-779
Copyright: © Gong et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Vascular endothelial growth factor (VEGF) and osteopontin (OPN) are suggested to facilitate angiogenesis and vascular remodeling in endometrial receptivity. Determination of the endometrial microvascular density (MVD) is the commonest method used to indirectly assess the levels of vasculogenesis and angiogenesis; however, the associations among VEGF, OPN and MVD remain unclear. Controlled ovarian hyperstimulation (COH) with the gonadotrophin‑releasing hormone agonist‑long protocol may impair endometrial receptivity, and Traditional Chinese Medicine (TCM) may exert therapeutic effects to relieve this impairment. The aim of the present study was to investigate the effects of COH on implantation biology and pregnancy outcome, and to explore the potential therapeutic role of the TCM Zi Dan Yin (ZDY). Female Sprague Dawley rats were divided into four groups: Control, COH, ZDY and COH + ZDY. On days 3, 4 and 5 of pregnancy (D3, D4 and D5, respectively), endometrial MVD was measured with cluster of differentiation 34 immunohistochemical detection, and VEGF and OPN protein and mRNA expression was detected through western blotting and reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) analysis. On D10, the average number of implantation sites was observed. Subsequent to conceiving and bearing newborn rats, the number of live births from each group was recorded. COH was shown to have adverse effects on implantation and pregnancy outcome. The MVD was found to be significantly increased in the COH group compared with that in the control, ZDY and COH+ZDY groups. The results of the protein and RT-qPCR analysis of VEGF and OPN revealed the same trend. Conversely, ZDY reversed the changes in endometrial MVD, VEGF and OPN, and was indicated to improve uterine receptivity and pregnancy outcome. No significant difference was observed among the control, ZDY and COH + ZDY groups. In conclusion, since the results for MVD and VEGF and OPN expression were consistent, MVD could be used as an alternative approach to identify the period of receptivity in rats.

View Figures

View References

1	Finn CA and Martin L: The control of implantation. J Reprod Fertil. 39:195–206. 1974. View Article : Google Scholar : PubMed/NCBI
2	Tabibzadeh S and Babaknia A: The signals and molecular pathways involved in implantation, a symbiotic interaction between blastocyst and endometrium involving adhesion and tissue invasion. Hum Reprod. 10:1579–1602. 1995. View Article : Google Scholar : PubMed/NCBI
3	Cakmak H and Taylor HS: Implantation failure: molecular mechanisms and clinical treatment. Hum Reprod Update. 17:242–253. 2011. View Article : Google Scholar :
4	Sharkey AM and Smith SK: The endometrium as a cause of implantation failure. Best Pract Res Clin Obstet Gynaecol. 17:289–307. 2003. View Article : Google Scholar : PubMed/NCBI
5	Psychoyos A: Hormonal control of ovoimplantation. Vitam Horm. 31:201–256. 1973. View Article : Google Scholar : PubMed/NCBI
6	Psychoyos A: Hormonal control of uterine receptivity for nidation. J Reprod Fertil Suppl. 17–28. 1976.PubMed/NCBI
7	Renfree MB and Shaw G: Diapause. Annu Rev Physiol. 62:353–375. 2000. View Article : Google Scholar : PubMed/NCBI
8	Oldberg A, Franzén A and Heinegård D: Cloning and sequence analysis of rat bone sialoprotein (osteopontin) cDNA reveals an Arg-Gly-Asp cell-binding sequence. Proc Natl Acad Sci USA. 83:8819–8823. 1986. View Article : Google Scholar : PubMed/NCBI
9	Ruoslahti E and Pierschbacher MD: New perspectives in cell adhesion: RGD and integrins. Science. 238:491–497. 1987. View Article : Google Scholar : PubMed/NCBI
10	Brown LF, Berse B, Van de Water L, et al: Expression and distribution of osteopontin in human tissues: widespread association with luminal epithelial surfaces. Mol Biol Cell. 3:1169–1180. 1992. View Article : Google Scholar : PubMed/NCBI
11	Nomura S, Wills AJ, Edwards DR, Heath JK and Hogan BL: Developmental expression of 2ar (osteopontin) and SPARC (osteonectin) RNA as revealed by in situ hybridization. J Cell Biol. 106:441–450. 1988. View Article : Google Scholar : PubMed/NCBI
12	Maglione D, Guerriero V, Viglietto G, et al: Two alternative mRNAs coding for the angiogenic factor, placenta growth factor (PlGF), are transcribed from a single gene of chromosome 14. Oncogene. 8:925–931. 1993.PubMed/NCBI
13	Rabbani M and Rogers PA: Role of vascular endothelial growth factor in endometrial vascular events before implantation in rats. Reproduction. 122:85–90. 2001. View Article : Google Scholar : PubMed/NCBI
14	Rockwell LC, Pillai S, Olson CE and Koos RD: Inhibition of vascular endothelial growth factor/vascular permeability factor action blocks estrogen-induced uterine edema and implantation in rodents. Biol Reprod. 67:1804–1810. 2002. View Article : Google Scholar : PubMed/NCBI
15	Krüssel JS, Casañ EM, Raga F, et al: Expression of mRNA for vascular endothelial growth factor transmembraneous receptors Flt1 and KDR, and the soluble recetor sflt in cycling human endometrium. Mol Hum Reprod. 5:452–458. 1999. View Article : Google Scholar
16	de Mouzon J, Goossens V, Bhattacharya S, et al; European IVF-monitoring (EIM) Consortium, for the European Society of Human Reproduction and Embryology (ESHRE). Assisted reproductive technology in Europe, 2006: results generated from European registers by ESHRE. Hum Reprod. 25:1851–1862. 2010. View Article : Google Scholar : PubMed/NCBI
17	Levi AJ, Drews MR, Bergh PA, Miller BT and Scott RT Jr: Controlled ovarian hyperstimulation does not adversely affect endometrial receptivity in in vitro fertilization cycles. Fertil Steril. 76:670–674. 2001. View Article : Google Scholar : PubMed/NCBI
18	Pattinson HA, Greene CA, Fleetham J and Anderson-Sykes SJ: Exogenous control of the cycle simplifies thawed embryo transfer and results in a pregnancy rate similar to that for natural cycles. Fertil Steril. 58:627–629. 1992.PubMed/NCBI
19	Pellicer A, Valbuena D, Cano F, Remohí J and Simón C: Lower implantation rates in high responders: evidence for an altered endocrine milieu during the preimplantation period. Fertil Steril. 65:1190–1195. 1996.PubMed/NCBI
20	Bourgain C and Devroey P: The endometrium in stimulated cycles for IVF. Hum Reprod Update. 9:515–522. 2003. View Article : Google Scholar
21	Dey SK, Lim H, Das SK, et al: Molecular cues to implantation. Endocr Rev. 25:341–373. 2004. View Article : Google Scholar : PubMed/NCBI
22	Efferth T, Li PC, Konkimalla VSB and Kaina B: From traditional Chinese medicine to rational cancer therapy. Trends Mol Med. 13:353–361. 2007. View Article : Google Scholar : PubMed/NCBI
23	Wen Z, Wang Z, Wang S, et al: Discovery of molecular mechanisms of traditional Chinese medicinal formula Si-Wu-Tang using gene expression microarray and connectivity map. PLoS One. 6:e182782011. View Article : Google Scholar : PubMed/NCBI
24	Wilcox AJ, Weinberg CR, O’Connor JF, et al: Incidence of early loss of pregnancy. New Engl J Med. 319:189–194. 1988. View Article : Google Scholar : PubMed/NCBI
25	Norwitz ER, Schust DJ and Fisher SJ: Implantation and the survival of early pregnancy. N Engl J Med. 345:1400–1408. 2001. View Article : Google Scholar
26	Zhong-zhi Q, Yang D, Yan-ze L, et al: Pharmacopoeia of the People’s Republic of China (2010 Edition): A Milestone in Development of China’s Healthcare. Chinese Herbal Medicines. 2:157–160. 2010.
27	Collins PD, Connolly DT and Williams TJ: Characterization of the increase in vascular permeability induced by vascular permeability factor in vivo. Br J Pharmacol. 109:195–199. 1993. View Article : Google Scholar : PubMed/NCBI
28	Hippenstiel S, Krüll M, Ikemann A, et al: VEGF induces hyperpermeability by a direct action on endothelial cells. Am J Physiol. 274:L678–L684. 1998.PubMed/NCBI
29	Goodger AM and Rogers PA: Uterine endothelial cell proliferation before and after embryo implantation in rats. J Reprod Fertil. 99:451–457. 1993. View Article : Google Scholar : PubMed/NCBI
30	Ramsey EM and Donner MW: Placental Vasculature and Circulation: Anatomy, Physiology, Radiology, Clinical Aspects: Atlas and Textbook. Thieme; Stuttgart: 1980
31	Jakeman LB, Armanini M, Phillips HS and Ferrara N: Developmental expression of binding sites and messenger ribonucleic acid for vascular endothelial growth factor suggests a role for this protein in vasculogenesis and angiogenesis. Endocrinology. 133:848–859. 1993.PubMed/NCBI
32	Breier G, Albrecht U, Sterrer S and Risau W: Expression of vascular endothelial growth factor during embryonic angiogenesis and endothelial cell differentiation. Development. 114:521–532. 1992.PubMed/NCBI
33	Giudice LC: Genes associated with embryonic attachment and implantation and the role of progesterone. J Reprod Med. 44(2 Suppl): 165–171. 1999.
34	Young MF, Kerr JM, Termine JD, et al: cDNA cloning, mRNA distribution and heterogeneity, chromosomal location, and RFLP analysis of human osteopontin (OPN). Genomics. 7:491–502. 1990. View Article : Google Scholar : PubMed/NCBI
35	Carson DD, Lagow E, Thathiah A, et al: Changes in gene expression during the early to mid-luteal (receptive phase) transition in human endometrium detected by high-density microarray screening. Mol Hum Reprod. 8:871–879. 2002. View Article : Google Scholar : PubMed/NCBI
36	Kao L, Tulac S, Lobo SA, et al: Global gene profiling in human endometrium during the window of implantation. Endocrinology. 143:2119–2138. 2002. View Article : Google Scholar : PubMed/NCBI
37	Borthwick JM, Charnock-Jones DS, Tom BD, et al: Determination of the transcript profile of human endometrium. Mol Hum Reprod. 9:19–33. 2003. View Article : Google Scholar : PubMed/NCBI
38	Talbi S, Hamilton A, Vo K, et al: Molecular phenotyping of human endometrium distinguishes menstrual cycle phases and underlying biological processes in normo-ovulatory women. Endocrinology. 147:1097–1121. 2006. View Article : Google Scholar
39	Chaen T, Konno T, Egashira M, et al: Estrogen-dependent uterine secretion of osteopontin activates blastocyst adhesion competence. PLoS One. 7:e489332012. View Article : Google Scholar : PubMed/NCBI
40	Liaw L, Birk DE, Ballas CB, Whitsitt JS, Davidson JM and Hogan BL: Altered wound healing in mice lacking a functional osteopontin gene (spp1). J Clin Invest. 101:1468–1478. 1998. View Article : Google Scholar : PubMed/NCBI
41	Rittling SR, Matsumoto HN, Mckee MD, et al: Mice lacking osteopontin show normal development and bone structure but display altered osteoclast formation in vitro. J Bone Miner Res. 13:1101–1111. 1998. View Article : Google Scholar : PubMed/NCBI
42	Liaw L, Almeida M, Hart CE, Schwartz SM and Giachelli CM: Osteopontin promotes vascular cell adhesion and spreading and is chemotactic for smooth muscle cells in vitro. Circ Res. 74:214–224. 1994. View Article : Google Scholar : PubMed/NCBI
43	Beaumont HM and Smith AF: Embryonic mortality during the pre- and post-implantation periods of pregnancy in mature mice after superovulation. J Reprod Fertil. 45:437–448. 1975. View Article : Google Scholar : PubMed/NCBI
44	Ertzeid G and Storeng R: Adverse effects of gonadotrophin treatment on pre- and postimplantation development in mice. J Reprod Fertil. 96:649–655. 1992. View Article : Google Scholar : PubMed/NCBI
45	McKiernan SH and Bavister BD: Gonadotrophin stimulation of donor females decreases post-implantation viability of cultured one-cell hamster embryos. Hum Reprod. 13:724–729. 1998. View Article : Google Scholar : PubMed/NCBI
46	Miller BG and Armstrong DT: Effects of a superovulatory dose of pregnant mare serum gonadotropin on ovarian function, serum estradiol, and progesterone levels and early embryo development in immature rats. Biol Reprod. 25:261–271. 1981. View Article : Google Scholar : PubMed/NCBI
47	Ertzeid G and Storeng R: The impact of ovarian stimulation on implantation and fetal development in mice. Hum Reprod. 16:221–225. 2001. View Article : Google Scholar : PubMed/NCBI
48	Foote R and Ellington J: Is a superovulated oocyte normal? Theriogenology. 29:111–123. 1988. View Article : Google Scholar
49	Allen J and McLaren A: Cleavage rate of mouse eggs from induced and spontaneous ovulation. J Reprod Fertil. 27:137–140. 1971. View Article : Google Scholar : PubMed/NCBI
50	Makrigiannakis A, Minas V, Kalantaridou SN, Nikas G and Chrousos GP: Hormonal and cytokine regulation of early implantation. Trends Endocrinol Metab. 17:178–185. 2006. View Article : Google Scholar : PubMed/NCBI
51	Ruan HC, Zhu XM, Luo Q, et al: Ovarian stimulation with GnRH agonist, but not GnRH antagonist, partially restores the expression of endometrial integrin beta3 and leukaemia-inhibitory factor and improves uterine receptivity in mice. Hum Reprod. 21:2521–2529. 2006. View Article : Google Scholar : PubMed/NCBI