Effects of Progesterone and 17β-Estradiol under Presence or Absence of FBS on Plasminogen Activators Activity in Porcine Uterine Epithelial Cells
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The present study was conducted to investigate the regulatory mechanism of plasminogen activators (PAs) ac-tivation by 17β-estradiol (E2) and progesterone (P4) in porcine uterine epithelial cells (pUECs). pUECs were collected from porcine uterine horn and cultured at 80% confluence. Then, 0.1% (v/v) DMSO, 20 ng/mL E2, and P4 with or without fetal bo-vine serum (FBS) treated to cultured cells for 24 hours. The supernatants were used for measurement of PAs activity and ex-pression of urokinase-type PA (uPA), tissue-type PA (tPA), uPA specific receptor (uPAR), and type-1 PA inhibitor (PAI-1) mRNA were analyzed by real-time PCR. The expression of PAs-related genes was not affect by steroid hormones in both of serum treatment groups. However, PAs activity was increased by treatment of E2 compared to 0.1% DMSO treatment in se-rum-free group (p<0.05). Then, E2 and P4 were diluted with 0.002% (v/v) DMSO for reduction of its effect and treated to cul-tured cells without FBS. Only tPA mRNA was significantly increased by E2 treatment (p<0.05). PAs activity was enhanced in E2 treated group compared to control groups (p<0.05). These results indicate that serum-free condition is more proper to eval-uate effect of steroid hormones and activation of PAs in pUECs was mainly regulated by estrogen. These regulation of PAs activation may be associated with uterine remodeling during pre-ovulatory phase in pigs, however, further studies are needed to investigate precise regulatory mechanism.
Abstract
The present study was conducted to investigate the regulatory mechanism of plasminogen activators (PAs) ac-tivation by 17β-estradiol (E2) and progesterone (P4) in porcine uterine epithelial cells (pUECs). pUECs were collected from porcine uterine horn and cultured at 80% confluence. Then, 0.1% (v/v) DMSO, 20 ng/mL E2, and P4 with or without fetal bo-vine serum (FBS) treated to cultured cells for 24 hours. The supernatants were used for measurement of PAs activity and ex-pression of urokinase-type PA (uPA), tissue-type PA (tPA), uPA specific receptor (uPAR), and type-1 PA inhibitor (PAI-1) mRNA were analyzed by real-time PCR. The expression of PAs-related genes was not affect by steroid hormones in both of serum treatment groups. However, PAs activity was increased by treatment of E2 compared to 0.1% DMSO treatment in se-rum-free group (p<0.05). Then, E2 and P4 were diluted with 0.002% (v/v) DMSO for reduction of its effect and treated to cul-tured cells without FBS. Only tPA mRNA was significantly increased by E2 treatment (p<0.05). PAs activity was enhanced in E2 treated group compared to control groups (p<0.05). These results indicate that serum-free condition is more proper to eval-uate effect of steroid hormones and activation of PAs in pUECs was mainly regulated by estrogen. These regulation of PAs activation may be associated with uterine remodeling during pre-ovulatory phase in pigs, however, further studies are needed to investigate precise regulatory mechanism.
참고문헌 (41)
- Shin-Hye Ahn, "Relationship between Plasminogen Activity and Plasminogen Inhibitor during the Culture of Porcine Oviduct Epithelial Cells", Reproductive & Developmental biology, 33(4), pp.203~209 (2009), 사단법인 한국동물생명공학회
- Baker VL, "Reproductive endocrine and endometrial effects of raloxifene hydrochloride, a se-lective estrogen receptor modulator, in women with regular menstrual cycles", J Clin Endocr Metab, 83, pp.6~13 (1998)
- Bazer FW, "Interferons and progesterone for establishment and maintenance of pregnancy: Interactions among novel cell signaling pathways", Reprod Biol, 8, pp.179~211 (2008)
- Bazer FW, "Pig blastocyst-uterine in-teractions", Differentiation, 87, pp.52~65 (2014)
- Beers WH, "Follicular plasminogen and plasminogen activator and the effect of plasmin on ovarian follicle wall", Cell, 6, pp.379~386 (1975)
- Casslen B, "Occurrence of both urokinase and tissue plasminogen activator in the human endo-metrium", Contraception, 28, pp.553~564 (1983)
- 전용필, "착상전 생쥐 자궁에서 콜라겐의 변화", 발생과 생식, 11(1), pp.1~11 (2007), 한국발생생물학회
- 전용필, "Androgen in the Uterus: A Compensator of Estrogen and Progesterone", 발생과 생식, 13(3), pp.133~143 (2009), 한국발생생물학회
- Coy P, "Oocytes use the plasmino-gen-plasmin system to remove supernumerary sperma-tozoa", Hum Reprod, 27, pp.1985~1993 (2012)
- de Boer JP, "Alpha-2-macroglobulin functions as an inhibitor of fibrinolytic, clotting, and neutrophilic proteinases in sepsis: studies using a baboon model", Infect Immun, 61, pp.5035~5043 (1993)
- Demir R, "Vasculogenesis and angiogenesis in the endometrium during menstrual cy-cle and implantation", Acta Histochem, 112, pp.203~214 (2010)
- Ebisch IMW, "Review of the role of the plasminogen activators sys-tem and vascular endothelial growth factor in subfertil-ity", Fertil Steril, 90, pp.2340~2350 (2008)
- Elangovan S, "SIRT1 is essential for oncogenic signaling by estrogen", estrogen receptor α in breast cancer, 2011, pp.71, 6654~6664
- Fazleabas AT, "Purification and properties of a progesterone-induced plasmin", tryp-sin inhibitor from uterine secretions of pigs and its immunocytochemical localization in the pregnant uter-us, 1982, pp.257, 6886~6897
- Finlay TH, "Estrogen-stimulated uptake of plasminogen by the mouse uterus", Endocrinology, 112, pp.856~861 (1983)
- Franczak A, "Tran-scriptomic analysis of the porcine endometrium during early pregnancy and the estrous cycle", Reprod Biol, 13, pp.229~237 (2013)
- Gray CA, "Developmental biology of uterine glands", Biol Reprod, 65, pp.1311~1323 (2001)
- 황보용, "돼지의 배란 전 자궁내막 상피세포 내 Plasminogen Activators의 발현", 한국동물생명공학회지, 28(3), pp.257~263 (2013), 사단법인 한국동물생명공학회
- Ka H, "Keratinocyte growth factor is up-regulated by estrogen in the porcine uterine endometrium and func-tions in trophectoderm cell proliferation and differenti-ation", Endocrinology, 142, pp.2303~2310 (2001)
- Kang-Hyun Kim, "Changes in Plasminogen Activity in Uterus Tissue during the Estrous Cycle in the Pigs", Reproductive & Developmental biology, 35(4), pp.463~468 (2011), 사단법인 한국동물생명공학회
- Kobayashi T, "Trypsin-like arginine amidases including plasminogen and plasmin in human seminal plasma by affinity adsorption and elution", Arch Androl, 28, pp.165~170 (1992)
- Kouba AJ, "Oviductal plasminogen activator in-hibitor-1 (PAI-1): mRNA, protein, and hormonal regu-lation during the estrous cycle and early pregnancy in the pig", Mol Reprod Dev, 56, pp.378~386 (2000)
- Krania F, "Effects of addition of tissue-type plasminogen activator in in vitro fertilization me-dium on bovine embryo development and quality", Re-prod Domest Anim, 50, pp.112~120 (2015)
- Lai MD, "Expres-sion of proliferating cell nuclear antigen in luminal epi-thelium during the growth and regression of rat uterus", J Endocrinol, 166, pp.87~93 (2000)
- Martin O, "Plasminogen activator produc-tion by trophoblast cells in vitro : Effect of steroid hor-mones and protein synthesis inhibitors", Am J Obstet Gynecol, 142, pp.402~409 (1982)
- McCracken JA, "Hormone receptor control of pros-taglandin F2 alpha secretion by the ovine uterus", Adv Prostaglandin Thromboxane Res, 8, pp.1329~1344 (1980)
- Menshikov M, "Urokinase plasminogen activator stimulates vascular smooth muscle cell proliferation via redox-dependent pathways", Arter Thromb Vasc Biol, 26, pp.801~807 (2006)
- Mullins DE, "Secretion of a progesterone-induced inhibitor of plasminogen activa-tor by the porcine uterus", Cell, 20, pp.865~872 (1980)
- Noguchi M, "Peripheral concentra-tions of inhibin A, ovarian steroids, and gonadotropins associated with follicular development throughout the estrous cycle of the sow", Reproduction, 139, pp.153~161 (2010)
- Olofsson B, "Vascular endothelial growth factor B (VEGF-B) binds to VEGF receptor-1 and reg-ulates plasminogen activator activity in endothelial cells", Proc Natl Acad Sci USA, 95, pp.11709~11714 (1998)
- Ploplis VA, "Plasminogen deficiency differentially affects recruitment of inflammatory cell populations in mice", Blood, 91, pp.2005~2009 (1998)
- Sappino AP, "Plas-minogen activators in tissue remodeling and invasion: mRNA localization in mouse ovaries and implanting embryos", J Cell Biol, 109, pp.2471~2479 (1989)
- Schatz F, "Plasminogen activator activity during decidualization of human endometrial stromal cells is regulated by plasminogen activator inhibitor 1", J Clin Endocrinol Metab, 80, pp.2504~2510 (1995)
- Schatz F, "Progestin regulation of plasminogen activator inhibitor type 1 in primary cul-tures of endometrial stromal and decidual cells", J Clin Endocrinol Metab, 77, pp.621~625 (1993)
- Spencer TE, "Biology of progesterone action during pregnancy recognition and maintenance of pregnancy", Front Biosci, 7, pp.1879~1898 (2002)
- Spencer TE, "Ovine interferon-tau regulates ex-pression of endometrial receptors for estrogen and oxy-tocin but not progesterone", Biol Reprod, 53, pp.732~745 (1995)
- Spencer TE, "Progesterone and placental hormone actions on the uter-us : Insights from domestic animals", Biol Reprod, 71, pp.2~10 (2004)
- Stroband HWJ, "The ultrastructure of the uterine epithelium of the pig during the estrous cycle and early pregnancy", Cell Tissue Res, 246, pp.81~89 (1986)
- Tanikawa M, "Cell-type specificity of interleukins 1α and 1β on prostaglandin and plasmino-gen activator production in bovine endometrial cell", Anim Reprod Sci, 114, pp.32~42 (2009)
- White FJ, "Steroid regulation of cell specific secreted phosphoprotein 1 (osteopontin) expression in the pregnant porcine uterus", Biol Reprod, 73, pp.1294~1301 (2005)
- Yan W, "Relaxin (RLX) and estrogen affect estrogen receptor α, vascular endothelial growth factor, and RLX receptor expression in the neonatal porcine uterus and cervix", Reproduction, 135, pp.705~712 (2008)
