17β-Estradiol Up-Regulates Prostacyclin Production in Cultured Human Uterine Myometrial Cells Via Augmentation of Both Cyclooxygenase-1 and Prostacyclin Synthase Expression

Abstract

Objective: To investigate whether 17β-estradiol elevates prostacyclin (PGI₂) production in human myometrial cells in the middle of gestation.

Methods: The concentration of 6-keto-PGF_1α, a stable metabolite of PGI₂, in the culture medium was assessed using enzyme-linked immunosorbent assay (ELISA). Western blot analysis and quantitative reverse-transcriptase polymerase chain reaction (RT-PCR) using TaqMan (Applied Biosystems, Foster City, CA) technology were performed to evaluate the expression of cytosolic phospholipase A₂ (cPLA₂), cyclooxygenase-1 (COX-1), COX-2, and prostacyclin synthase (PGIS) in cultured human myometrial cells prepared from second trimester pregnant women (n = 3) after stimulation with 17β-estradiol.

Results: Treatment with 17β-estradiol (4-400 nM) dose-dependently elevated PGI₂ secretion from cultured human myometrial cells. Western blot analysis detected cPLA₂ and COX-1 and PGIS protein expression in the cultured human myometrial cells; however, COX-2 protein expression was below the detection sensitivity. Stimulation with 40-nM 17β-estradiol significantly up-regulated protein and mRNA expression of both COX-1 and PGIS.

Conclusion: 17βEstradiol from placenta may contribute to the augmentation of PGI₂ production in the human myometrium in the middle of gestation via up-regulation of both COX-1 and PGIS expression.

Keywords

Estrogen myometrium pregnancy prostacyclin prostacyclin synthase

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References

1. Okazaki T , Casey ML , Okita JR , MacDonald PC , Johnston JM . Initiation of human parturition. XII. Biosynthesis and metabolism of prostaglandins in human fetal membranes and uterine decidua. Am J Obstet Gynecol 1981;139:373-381.

2. Challis JR , Lye SJ , Gibb W . Prostaglandins and parturition. Ann N Y Acad Sci 1997;828:254-267.

3. Gibb W . The role of prostaglandins in human parturition. Ann Med 1998;30:235-241.

4. Cunningham FG , Gant NF , Leveno KJ , Gilstrap LC , Hauth JC , Wenstrom KD . Parturition. In: Williams Obstetrics. 21st ed. Norwalk, CT: McGraw-Hill, 2001:251-290.

5. Wilhelmsson L , Wikland M , Wiqvist N . PGH2, TXA2 and PGI2 have potent and differentiated actions on human uterine contractility. Prostaglandins 1981;21:277-286.

6. Matsumoto T , Sagawa N , Yoshida M , et al. The prostaglandin E2 and F2α receptor genes are expressed in human myometrium and are down-regulated during pregnancy. Biochem Biophys Res Commun 1997;238:838-841.

7. Magness RR , Shideman CR , Habermehl DA , Sullivan JA , Bird IM . Endothelial vasodilator production by uterine and systemic arteries. V. Effects of ovariectomy, the ovarian cycle, and pregnancy on prostacyclin synthase expression. Prostaglandins Other Lipid Mediat 2000;60:103-118.

8. Lumsden MA , Baird DT . The effect of intrauterine administration of prostacyclin on the contractility of the nonpregnant uterus in vivo. Prostaglandins 1986;31:1011-1022.

9. Dyal R , Crankshaw DJ . The effects of some synthetic prostanoids on the contractility of the human lower uterine segment in vitro. Am J Obstet Gynecol 1988;158:281-285.

10.

10. Abel MH , Kelly RW . Differential production of prostaglandins within the human uterus. Prostaglandins 1979;18:821-828.

11.

11. Toddz HM , Dundoo, VL , Gerber WR , Cwiak CA , Baldassare JJ , Hertelendy F . Effect of cytokines on prostaglandin E2 and prostacyclin production in primary cultures of human myometrial cells. J Matern Fetal Med 1996;5:161-167.

12.

12. Christensen NJ , Green K . Bioconversion of arachidnic acid in human pregnant reproductive tissues. Biochem Med 1983;30:162-180.

13.

13. Moonen P , Klok G , Keirse MJ . Distribution of prostaglandin endoperoxide synthase and prostacyclin synthase in the late pregnant uterus. Br J Obstet Gynaecol 1986;93:255-259.

14.

14. Wang YP , Walsh SW , Guo JD , Zhang JY . Maternal levels of prostacyclin, thromboxane, vitamin E and lipid peroxides throughout normal pregnancy. Am J Obstet Gynecol 1991;165:1690-1694.

15.

15. Goodman RP , Killam AP , Brash AR , Branch RA . Prostacyclin production during pregnancy: Comparison of production during normal pregnancy and pregnancy complicated by hypertension. Am J Obstet Gynecol 1982;142:817-822.

16.

16. Bamford DS , Jogee M , Williams KI . Prostacyclin formation by the pregnant human myometrium. Br J Obstet Gynaecol 1980;87:215-218.

17.

17. Ylikorkala O , Makarainen L , Viinikka L . Prostacyclin production increases during human parturition. Br J Obstet Gynaecol 1981;88:513-516.

18.

18. Davidson BJ , Murray RD , Challis JR , Valenzuela GJ . Estrogen, progesterone, prolactin, prostaglandin E2, prostaglandin F2α, 13, 14-dihydro-15-keto-prostaglandin F2μα and 6-keto-prostaglandin F1α gradients across the uterus in women in labor and not in labor. Am J Obstet Gynecol 1987;157:54-58.

19.

19. Ganong WF . Energy balance, metabolism, and nutrition. In: Ganong WF , ed. Review of Medical Physiology. 20th ed. New York: McGraw-Hill, 2001;271-306.

20.

20. Miyata A , Hara S , Yokoyama C , Inoue H , Ullrich V , Tanabe T . Molecular cloning and expression of human prostacyclin synthase. Biochem Biophys Res Commun 1994;200:1728-1734.

21.

21. Korita D , Sagawa N , Itoh H , et al. Cyclic mechanical stretch augments prostacyclin production in cultured human uterine myometrial cells from pregnant women: Possible involvement of up-regulation of prostacyclin synthase expression. J Clin Endocrinol Metab 2002;87:5209-5219.

22.

22. Korita D , Itoh H , Sagawa N , et al. Cyclic mechanical stretch and interleukin-1α synergistically up-regulate prostacyclin secretion in cultured human uterine myometrial cells. Gynecol Endocrinol 2004;18:130-137.

23.

23. Speroff L , Glass RH , Kase NG . The Endocrinology of Pregnancy. In: Clinical Gynecologic Endocrinology and Infertility. 6th ed. Baltimore: Lippincott Williams & Wilkins, 1999:275-235.

24.

24. Seillen C , Ody C , Russo-Marie F , Duval D . Differential effects of sex steroids on prostaglandin secretion by male and female cultured piglet endothelial cells. Prostaglandins 1983;26:3-12.

25.

25. Chang WC , Nakao J , Orimo H , Murota S . Stimulation of prostacyclin biosynthetic activity by estradiol in rat aortic smooth muscle cells in culture. Biochim Biophys Acta 1980;620:472-482.

26.

26. Jun SS , Chen Z , Pace MC , Shaul PW . Estrogen up-regulates cyclooxygenase-1 gene expression in ovine fetal pulmonary artery endothelium. J Clin Invest 1998;102:176-183.

27.

27. Rupnow HL , Phernetton TM , Modrick ML , Wiltbank MC , Bird IM , Magness RR . Endothelial vasodilator production by uterine and systemic arteries. VIII. Estrogen and progesterone effects on cPLA 2, COX-1, and PGIS protein expression. Biol Reprod 2002;66:468-474.

28.

28. Ospina JA , Duckles SP , Krause DN . 17Beta-estradiol decreases vascular tone in cerebral arteries by shifting COX-dependent vasoconstriction to vasodilation. Am J Physiol Heart Circ Physiol 2003;285:H241-H250.

29.

29. Kakui K , Itoh H , Sagawa N , et al. Augmented eNOS protein expression in human pregnant myometrium: Possible involvement of eNOS promoter activation by estrogen via both ERα and ERβ. Mol Hum Reprod 2004;10:115-122.

30.

30. Devroey P , Camus M , Palermo G , et al. Placentral production of estradiol and progesterone after oocyte donation in donation in patients with primary ovarian failure. Am J Obstet Gynecol 1990;162:66-70.

31.

31. Dong YL , Wimalawana S , Yallampalli C . Effects of steroid hormones on calcitonin gene-related peptide receptors in cultured human myometrium. Am J Obstet Gynecol 2003;188:466-472.

32.

32. Fiorini s , Ferretti ME , Biondi C , et al. 17β-Estradiol stimulates arachidonate release from human amnion-like WISH cells through a rapid mechanism involving a membrane receptor. Endocrinology 2003;144:3359-3367.

33.

33. Pavan B , Biondi C , Ferretti ME , Lunghi L , Paganetto G . 17β-Estradiol modulates prostaglandin E2 release from human amnion-derived wish cells. Biol Reprod 2001;64:1677-1681.

34.

34. Terakawa K , Itoh H , Sagawa N , et al. Site-specific augmentation of amnion cyclooxygenase-2 and desidua vera phospholipase-A2 expression in labor: possible contribution of mechanical stretch and interleukin-1 to amnion prostaglandin synthesis. J Soc Gynecol Investig 2002;9:68-74.

35.

35. Masuzaki H , Ogawa Y , Sagawa N , et al. Nonadipose tissue production of leptin: Leptin as a novel placenta-derived hormone in humans. Nat Med 1997;3:1029-1033.

36.

36. Yoshida M , Sagawa N , Itoh H , et al. Nitric oxide increases matrix metalloproteinase-1 production in human uterine cervical fibroblast cells. Mol Hum Reprod 2001;7:979-985.

37.

37. Kakui K , Sagawa N , Itoh H , et al. Expression of nitric oxide synthase isoforms in the human placenta is not altered by labor. Endocrine J 2003;50:535-544.

38.

38. Yokoyama C , Tanabe T . Cloning of human gene encoding prostaglandin endoperoxide synthase and primary structure of the enzyme. Biochem Biophys Res Commun 1989;165:888-894.

39.

39. Tso JY , Sun XH , Kao TH , Reece KS , Wu R . Isolation and characterization of rat and human glyceraldehyde-3-phosphate dehydrogenase cDNAs: Genomic complexity and molecular evolution of the gene. Nucleic Acids Res 1985;13:2485-2502.

40.

40. Gargett CE , Bucak K , Zaitseva M , et al. Estrogen receptor-alpha and -beta expression in microvascular endothelial cells and smooth muscle cells of myometrium and leiomyoma. Mol Hum Reprod 2002;8:770-775.

41.

41. Gustafsson JA . Estrogen receptor β—A new dimension in estrogen mechanism of action. J Endocrinol 1999;163:379-383.

42.

42. Xu XM , Tang JL , Chen X , Wang LH , Wu KK . Involvement of two Sp1 elements in basal endothelial prostaglandin H synthase-1 promoter activity. J Biol Chem 1997;272:6943-6950.

43.

43. Wang LH , Chen L . Organization of the gene encoding human prostacyclin synthase. Biochem Biophys Res Commun 1996;226:631-637.

44.

44. Kato S , Tora L , Yamauchi J , Masushige S , Bellard M , Chambon P . A far upstream estrogen response element of the ovalbumin gene contains several half palindromic 5′-TGACC-3′ motifs acting synergistically. Cell 1992;68:731-742.

45.

45. L'Horset F , Blin C , Colnot S , Lambert M , Thomasset M , Perret C . Calbindin-D9k gene expression in the uterus: Study of two messenger ribonucleic acid species and analysis of an imperfect estrogen responsive element. Endocrinology 1994;134:11-18.

46.

46. Murdoch FE , Byrne LM , Anazi EA , Furlow DJ , Meier DA , Gorski J . Estrogen receptor binding to DNA: affinity for non-palindromic elements from the rat prolactin gene. Biochemistry 1995;34:9144-9150.

47.

47. Parker TA , Afshar S , Kinsella JP , et al. Effects of chronic estrogen-receptor blockade on ovine perinatal pulmonary circulation. Am J Physiol Heart Circ Physiol 2001;281:H1005-H1014.

48.

48. Enmark E , Gustafsson JA . Oestrogen receptors—An overview. J Intern Med 1999;246:133-138.

49.

49. Mendelsohn ME , Karas RH . The protective effects of estrogen on the cardiovascular system. N Engl J Med 1999;340:1801-1811.

50.

50. Itoh H , Sagawa N , Matsumoto T , et al. Role of the guanylate cyclase system in the regulation of ovine uterine arterial blood flow during pregnancy, a review. Trophoblast Res 1999;13:301-309.

51.

51. Rosenfeld CR , Cox BE , Roy T , Magness RR . Nitric oxide contributes to estrogen-induced vasodilation of the ovine uterine circulation. J Clin Invest 1996;98:2158-2166.

52.

52. Lim H , Dey SK . A novel pathway of prostacyclin signaling-hanging out with nuclear receptors. Endocrinology 2002;143:3207-3210.

53.

53. Hatae T , Wada M , Yokoyama C , Shimonishi M , Tanabe T . Prostacyclin-dependent apoptosis mediated by PPAR delta. J Biol Chem 2001;276:46260-46267.