Vol 55, No 3 (2017)
Original paper
Published online: 2017-10-09
The impact of antiandrogen flutamide on the hypoxia inducible factor 1a and vascular endothelial growth factor A gene and protein expression in the pig placenta during late pregnancy
Abstract
Introduction. In contrast to estradiol action, little is known about androgen signaling in placental development. The purpose of this study was to evaluate the impact of diminished androgen action on hypoxia inducible factor 1a (HIF-1a) and vascular endothelial growth factor A (VEGFA) protein expression as well as their mRNAs in the structures of fetal and maternal parts of porcine placenta during late pregnancy.
Material and methods. Pregnant pigs were injected daily with antiandrogen flutamide, at a dose of 50 mg/kg body weight at different stages of pregnancy: between gestational days 83–89 (90 dpc) and 101–107 (108 dpc). Control groups (90 dpc or 108 dpc) were treated with vehicle (corn oil). One day after the last injection animals were sacrificed and tissues were collected. Tissue samples were frozen for mRNA isolation or fixed for immunohistochemistry (IHC). The expression of HIF-1a and VEGFA were investigated by real-time PCR and IHC.
Results. Flutamide treatment caused changes in both HIF-1a and VEGFA mRNA levels only in the placentas of the 90 dpc group. Relative optical density analysis showed decreased HIF-1a and increased VEGFA protein expression in the placentas obtained from flutamide-treated 108 dpc group while no differences were observed in the 90 dpc group.
Conclusions. Experimentally induced androgen deficiency in pigs deregulates the expression of some genes important for placental blood circulation. We suggest that androgens are involved in the control of expression of HIF-1a and VEGFA in porcine placenta during late pregnancy.
Keywords: VEGFAHIF-1aandrogensflutamideplacentapigIHCQPCR
References
- Maliqueo M, Echiburú B, Crisosto N. Sex Steroids Modulate Uterine-Placental Vasculature: Implications for Obstetrics and Neonatal Outcomes. Front Physiol. 2016; 7: 152.
- Wang Q, Shen L, Huang W, et al. Vasculogenesis of decidua side population cells of first-trimester pregnancy. Stem Cell Res Ther. 2013; 4(3): 50.
- Albrecht ED, Pepe GJ. Estrogen regulation of placental angiogenesis and fetal ovarian development during primate pregnancy. Int J Dev Biol. 2010; 54(2-3): 397–408.
- Robb VA, Pepe GJ, Albrecht ED. Acute temporal regulation of placental vascular endothelial growth/permeability factor expression in baboons by estrogen. Biol Reprod. 2004; 71(5): 1694–1698.
- Corcoran JJ, Nicholson C, Sweeney M, et al. Human uterine and placental arteries exhibit tissue-specific acute responses to 17β-estradiol and estrogen-receptor-specific agonists. Mol Hum Reprod. 2014; 20(5): 433–441.
- Cheng G, Weihua Z, Mäkinen S, et al. A role for the androgen receptor in follicular atresia of estrogen receptor beta knockout mouse ovary. Biol Reprod. 2002; 66(1): 77–84.
- Knapczyk-Stwora K, Grzesiak M, Duda M, et al. TGFβ (transforming growth factor β) superfamily members and their receptors in the fetal porcine ovaries: effect of prenatal flutamide treatment. Folia Histochem Cytobiol. 2014; 52(4): 317–325.
- Hsu TY, Lan KC, Tsai CC, et al. Expression of androgen receptor in human placentas from normal and preeclamptic pregnancies. Taiwan J Obstet Gynecol. 2009; 48(3): 262–267.
- Wieciech I, Durlej-Grzesiak M, Słomczyńska M. Influence of the antiandrogen flutamide on the androgen receptor gene expression in the placenta and umbilical cord during pregnancy in the pig. Acta Histochem. 2013; 115(3): 290–295.
- Khatri P, Hoffmann B, Schuler G. Androgen receptor is widely expressed in bovine placentomes and up-regulated during differentiation of bovine trophoblast giant cells. Placenta. 2013; 34(5): 416–423.
- James JL, Stone PR, Chamley LW. The regulation of trophoblast differentiation by oxygen in the first trimester of pregnancy. Hum Reprod Update. 2006; 12(2): 137–144.
- Wang GL, Jiang BH, Rue EA, et al. Hypoxia-inducible factor 1 is a basic-helix-loop-helix-PAS heterodimer regulated by cellular O2 tension. Proc Natl Acad Sci U S A. 1995; 92(12): 5510–5514.
- Patel J, Landers K, Mortimer RH, et al. Regulation of hypoxia inducible factors (HIF) in hypoxia and normoxia during placental development. Placenta. 2010; 31(11): 951–957.
- Krock BL, Skuli N, Simon MC. Hypoxia-induced angiogenesis: good and evil. Genes Cancer. 2011; 2(12): 1117–1133.
- Zhou Y, McMaster M, Woo K, et al. Vascular endothelial growth factor ligands and receptors that regulate human cytotrophoblast survival are dysregulated in severe preeclampsia and hemolysis, elevated liver enzymes, and low platelets syndrome. Am J Pathol. 2002; 160(4): 1405–1423.
- Wieciech I, Grzesiak M, Knapczyk-Stwora K, et al. Influence of the antiandrogen flutamide on connexin 43 (Cx43) gene and protein expression in the porcine placenta and uterus during pregnancy. Folia Biol (Krakow). 2014; 62(4): 367–375.
- Knapczyk-Stwora K, Grzesiak M, Duda M, et al. Effect of flutamide on folliculogenesis in the fetal porcine ovary – regulation by Kit ligand/c-Kit and IGF1/IGF1R systems. Anim Reprod Sci. 2013; 142(3-4): 160–167.
- Gopalakrishnan K, Mishra JS, Chinnathambi V, et al. Elevated testosterone reduces uterine blood flow, spiral artery elongation, and placental oxygenation in pregnant rats. Hypertension. 2016; 67(3): 630–639.
- Santos TC, Oliveira MF, Papa PC, et al. VEGF system expression by immunohistochemistry and real-time RT-PCR study on collared peccary placenta. Theriogenology. 2014; 82(6): 834–843.
- Knapczyk-Stwora K, Belej A, Grzesiak M, et al. Effect of gestational antiandrogen treatment on Dicer1 expression in the porcine fetal gonads. Acta Histochem. 2015; 117(8): 725–731.
- Smolen AJ. Image analytic techniques for quantification of immunocytochemical staining in the nervous system. In: Conn PM. ed. Methods in neurosciences. Academic Press, New York 1990: 208–229.
- Pugh CW, Ratcliffe PJ. Regulation of angiogenesis by hypoxia: role of the HIF system. Nat Med. 2003; 9(6): 677–684.
- Cleys ER, Halleran JL, Enriquez VA, et al. Androgen receptor and histone lysine demethylases in ovine placenta. PLoS One. 2015; 10(2): e0117472.
- Palacios S. Androgens and female sexual function. Maturitas. 2007; 57(1): 61–65.
- Słomczyńska M, Duda M, Burek M, et al. Distribution of androgen receptor in the porcine uterus throughout pregnancy. Reprod Domest Anim. 2008; 43(1): 35–41.
- Gao W, Kim J, Dalton JT. Pharmacokinetics and pharmacodynamics of nonsteroidal androgen receptor ligands. Pharm Res. 2006; 23(8): 1641–1658.
- Dean A, Smith LB, Macpherson S, et al. The effect of dihydrotestosterone exposure during or prior to the masculinization programming window on reproductive development in male and female rats. Int J Androl. 2012; 35(3): 330–339.
- Morisset AS, Dubé MC, Drolet R, et al. Androgens in the maternal and fetal circulation: association with insulin resistance. J Matern Fetal Neonatal Med. 2013; 26(5): 513–519.
- Battaglia C, Mancini F, Fabbri R, et al. Vascular risk in young women with polycystic ovary and polycystic ovary syndrome. Obstet Gynecol. 2008; 111(2 Pt 1): 385–395.
- Fernandez EV, Reece KM, Ley AM, et al. Dual targeting of the androgen receptor and hypoxia-inducible factor 1α pathways synergistically inhibits castration-resistant prostate cancer cells. Mol Pharmacol. 2015; 87(6): 1006–1012.
- Saharinen P, Eklund L, Pulkki K, et al. VEGF and angiopoietin signaling in tumor angiogenesis and metastasis. Trends Mol Med. 2011; 17(7): 347–362.
- Günter J, Ruiz-Serrano A, Pickel C, et al. The functional interplay between the HIF pathway and the ubiquitin system – more than a one-way road. Exp Cell Res. 2017; 356(2): 152–159.
- Samanta D, Semenza GL. Maintenance of redox homeostasis by hypoxia-inducible factors. Redox Biol. 2017; 13: 331–335.
- Schönenberger MJ, Kovacs WJ. Hypoxia signaling pathways: modulators of oxygen-related organelles. Front Cell Dev Biol. 2015; 3: 42.
- Lazarus A, Keshet E. Vascular endothelial growth factor and vascular homeostasis. Proc Am Thorac Soc. 2011; 8(6): 508–511.
- Elks PM, Renshaw SA, Meijer AH, et al. Exploring the HIFs, buts and maybes of hypoxia signalling in disease: lessons from zebrafish models. Dis Model Mech. 2015; 8(11): 1349–1360.
- Oladipupo S, Hu S, Kovalski J, et al. VEGF is essential for hypoxia-inducible factor-mediated neovascularization but dispensable for endothelial sprouting. Proc Natl Acad Sci USA. 2011; 108(32): 13264–13269.
- Dietrich W, Gaba A, Zhegu Z, et al. Testosterone dependent androgen receptor stabilization and activation of cell proliferation in primary human myometrial microvascular endothelial cells. Fertil Steril. 2011; 95(4): 1247–12455.
- Berisha B, Schams D, Rodler D, et al. Expression pattern of HIF1alpha and vasohibins during follicle maturation and corpus luteum function in the bovine ovary. Reprod Dom Anim. 2016; 52(1): 130–139.
- Duda M, Burek M, Galas J, et al. Immunohistochemical localization of androgen receptor and aromatase in the ovary of the pregnant pig. Reprod Biol. 2004; 4(3): 289–298.
