Immunohistochemical (Ki-67) study of endometrial maturation in mice after use of phosphodiesterase type 5 inhibitor

Rashidi, Bahman; Rad, Jafar Soleimani; Rad, Leila Roshangar

Immunohistochemical (Ki-67) study of endometrial maturation in mice after use of phosphodiesterase type 5 inhibitor

Authors

¹ Department of Anatomical Sciences and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

² Department of Anatomy and Histology, School of Medicine and Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran

Abstract

Background : Uterine receptivity for the implantation is a complicated process, that ovarian factors (hormonal), endometrium and embryo simultaneously are involved in this phenomenon. A successful implantation needs appropriate development of the endometrium. Furthermore, embryo must be capable of reacting with the endometrium and producing suitable adhesion molecules. This study aimed to examine one of endometrial maturation indices in mice before implantation, i.e., proliferation of stromal cells.
Materials and Methods: A total of 40 adult female mice were divided into four groups: Control, gonadotropin, gonadotropin + progesterone, and gonadotropin + sildenafil citrate. The three experimental groups were first injected 7.5 IU of human chorionic gonadotropin (HCG) and then 7.5 IU of human menopausal gonadotropin (HMG). Then, every two female mice were placed in a cage with a male mouse for mating. Two groups were injected 1 mg of progesterone and 3 mg/kg of sildenafil citrate at intervals of 24, 48, and 72 h after injection of HMG. After 96 h, all the mice were killed, and their uterine samples subjected to tissue passage and prepared for analysis. Immunohistochemical method, Ki-67, and stromal mitotic cell count were used in this study.
Results: Our observations in all groups showed changes in the luminal epithelium. ANOVA analysis Ki-67-positive stromal cells among all groups were not statistically significant.
Conclusion: The results showed that administration of HMG and HCG following that of progesterone and sildenafil citrate could change the indices of endometrial maturation, and they were not involved in the phase immediately before implantation in stromal mitotic index.

Keywords

References

1.	Diedrich K, Fauser BC, Devroey P, Griesinger G, Evian Annual Reproduction (EVAR) Workshop Group. The role of the endometrium and embryo in human implantation. Hum Reprod Update 2007;13:365-77.
2.	Guillomot M. Cellular interactions during implantation in domestic ruminants. J Reprod Fertil Suppl 1995;49:39-51.
3.	Blankenship TN, Enders AC, King BF. Trophoblastic invasion and the development of uteroplacental arteries in the macaque: Immunohistochemical localization of cytokeratins, desmin, type IV collagen, laminin, and fibronectin. Cell Tissue Res 1993;272:227-36.
4.	Lessey BA, Castelbaum AJ. Integrins and implantation in the human. Rev Endocr Metab Disord 2002;3:107-17.
5.	Lessey BA. Endometrial responsiveness to steroid hormones: A moving target. J Soc Gynecol Investig 2004;11:61-2.
6.	Moyer DL, de Lignieres B, Driguez P, Pez JP. Prevention of endometrial hyperplasia by progesterone during long-term estradiol replacement: Influence of bleeding pattern and secretory changes. Fertil Steril 1993;59:992-7.
7.	Gonen Y, Casper RF. Prediction of implantation by the sonographic appearance of the endometrium during controlled ovarian stimulation for in vitro fertilization (IVF).t J In Vitro Fert Embryo Transf 1990;7:146-52.
8.	Sher G, Herbert C, Maassarani G, Jacobs MH. Assessment of the late proliferative phase endometrium by ultrasonography in patients undergoing in-vitro fertilization and embryo transfer (IVF/ET). Hum Reprod 1991;6:232-7.
9.	Amit A, Thaler I, Paz Y, Itskovitz-Eldor J. The effect of a nitric oxide donor on Doppler flow velocity waveforms in the uterine artery during the first trimester of pregnancy. Ultrasound Obstet Gynecol 1998;11:94-8.
10.	Cameron IT, Campbell S. Nitric oxide in the endometrium. Hum Reprod Update 1998;4:565-9.
11.	Sher G, Fisch JD. Vaginal sildenafil (Viagra): A preliminary report of a novel method to improve uterine artery blood flow and endometrial development in patients undergoing IVF. Hum Reprod 2000;15:806-9.
12.	Boolell M, Gepi-Attee S, Gingell JC, Allen MJ. Sildenafil, a novel effective oral therapy for male erectile dysfunction. Br J Urol 1996;78:257-61.
13.	Birner P, Ritzi M, Musahl C, Knippers R, Gerdes J, Voigtländer T, et al. Immunohistochemical detection of cell growth fraction in formalin-fixed and paraffin-embedded murine tissue. Am J Pathol 2001;158:1991-6.
14.	Ito T, Mitui H, Udaka N, Hayashi H, Okudela K, Kanisawa M, et al. Ki-67 (MIB 5) immunostaining of mouse lung tumors induced by 4-nitroquinoline 1-oxide. Histochem Cell Biol 1998;110:589-93.
15.	Gerlach C, Golding M, Larue L, Alison MR, Gerdes J. Ki-67 immunoexpression is a robust marker of proliferative cells in the rat. Lab Invest 1997;77:697-8.
16.	Scholzen T, Gerdes J. The Ki-67 protein: From the known and the unknown. J Cell Physiol 2000;182:311-22.
17.	Verheijen R, Kuijpers HJ, Schlingemann RO, Boehmer AL, van Driel R, Brakenhoff GJ, et al. Ki-67 detects a nuclear matrix-associated proliferation-related antigen. I. Intracellular localization during interphase. J Cell Sci 1989;92:123-30.
18.	Verheijen R, Kuijpers HJ, van Driel R, Beck JL, van Dierendonck JH, Brakenhoff GJ, et al. Ki-67 detects a nuclear matrix-associated proliferation-related antigen. II. Localization in mitotic cells and association with chromosomes. J Cell Sci 1989;92:531-40.
19.	Bruno S, Darzynkiewicz Z. Cell cycle dependent expression and stability of the nuclear protein detected by Ki-67 antibody in HL-60 cells. Cell Prolif 1992;25:31-40.
20.	Hall PA, McKee PH, Menage HD, Dover R, Lane DP. High levels of p53 protein in UV-irradiated normal human skin. Oncogene 1993;8:203-7.
21.	Salmi A, Heikkilä P, Lintula S, Rutanen EM. Cellular localization of c-jun messenger ribonucleic acid and protein and their relation to the proliferation marker Ki-67 in the human endometrium. J Clin Endocrinol Metab 1998;83:1788-96.
22.	Johns Hopkins Medical Institutions David March: Viagra (Sildenafil) effectively treats enlarged hearts, mouse study show. Johns Hopkins University: Medical News Today cardiology news; 2005. p. 24.
23.	Wareing M, Myers JE, O'Hara M, Baker PN. Sildenafil citrate (Viagra) enhances vasodilatation in fetal growth restriction. J Clin Endocrinol Metab 2005;90:2550-5.
24.	Khan RN, Hamoud H, Warren A, Wong LF, Arulkumaran S. Relaxant action of sildenafil citrate (Viagra) on human myometrium of pregnancy. Am J Obstet Gynecol 2004;191:315-21.
25.	Paulus WE, Strehler E, Zhang M, Jelinkova L, El-Danasouri I, Sterzik K. Benefit of vaginal sildenafil citrate in assisted reproduction therapy. Fertil Steril 2002;77:846-7.
26.	Rashidi B, Roshangar L, Soleimani Rad J, Khaki AA, Mohammadnejad D, Azami I. Comparison of morphology and morphometry of preimplantation mouse uterine endometrium in natural cycle with those received superovulatory drugs, progesterone and sildenafil citrate (Viagra). Pharm Sci Autumn, 2008; Vol 3, 33-39.
27.	Rashidi B, Rad JS, Roshangar L. Comparison of morphological and morphometrical characteristics in the glandular epithelium of mouse endometrium in preimplantation period after administration HMG-HCG, progesterone and sildenafil citrate. J Isfahan Med School 2010;28:112.
28.	Finn CA, Martin L. The role of the oestrogen secreted before oestrus in the preparation of the uterus for implantation in the mouse. J Endocrinol 1970;47:431-8.
29.	Huet-Hudson YM, Andrews GK, Dey SK. Cell type-specific localization of c-myc protein in the mouse uterus: Modulation by steroid hormones and analysis of the periimplantation period. Endocrinology 1989;125:1683-90.
30.	Gerdes J, Schwab U, Lemke H, Stein H. Production of a mouse monoclonal antibody reactive with a human nuclear antigen associated with cell proliferation. Int J Cancer 1983;31:13-20.
31.	Gerdes J, Lemke H, Baisch H, Wacker HH, Schwab U, Stein H. Cell cycle analysis of a cell proliferation-associated human nuclear antigen defined by the monoclonal antibody Ki-67. J Immunol 1984;133:1710-5.
32.	Gratzner HG. Monoclonal antibody to 5-bromo- and 5-iododeoxyuridine: A new reagent for detection of DNA replication. Science 1982;218:474-5.
33.	Alison MR. Assessing cellular proliferation: What's worth measuring? Hum Exp Toxicol 1995;14:935-44.
34.	Miyachi K, Fritzler MJ, Tan EM. Autoantibody to a nuclear antigen in proliferating cells. J Immunol 1978;121:2228-34.
35.	Foley J, Ton T, Maronpot R, Butterworth B, Goldsworthy TL. Comparison of proliferating cell nuclear antigen to tritiated thymidine as a marker of proliferating hepatocytes in rats. Environ Health Perspect 1993;101:199-205.
36.	Strauss JF, Gurpide E. The endometrium. In: Yen SS, Jaffe RB, editors. Reproductive Endocrinology. 3 ^rd ed. Philadelphia: W B Saunders; 1991. p. 309-56.
37.	Koshiyama M, Konishi I, Nanbu K, Nanbu Y, Mandai M, Komatsu T, et al. Immunohistochemical localization of heat shock proteins HSP70 and HSP90 in the human endometrium: Correlation with sex steroid receptors and Ki-67 antigen expression. J Clin Endocrinol Metab 1995;80:1106-12.
38.	Lessey BA, Killam AP, Metzger DA, Haney AF, Greene GL, McCarty KS Jr. Immunohistochemical analysis of human uterine estrogen and progesterone receptors throughout the menstrual cycle. J Clin Endocrinol Metab 1988;67:334-40.
39.	Garcia E, Bouchard P, De Brux J, Berdah J, Frydman R, Schaison G, et al. Use of immunocytochemistry of progesterone and estrogen receptors for endometrial dating. J Clin Endocrinol Metab 1988;67:80-7.
40.	Li SF, Nakayama K, Masuzawa H, Fujii S. The number of proliferating cell nuclear antigen positive cells in endometriotic lesions differs from that in the endometrium. Analysis of PCNA positive cells during the menstrual cycle and in post-menopause. Virchows Arch A Pathol Anat Histopathol 1993;423:257-63.
41.	Tabibzadeh S. Proliferative activity of lymphoid cells in human endometrium throughout the menstrual cycle. J Clin Endocrinol Metab 1990;70:437-43.

Advanced Biomedical Research