Objective:To investigate the effects of three cytokines, interleukin-1α (IL-1α), epidermal growth factor (EGF), and transforming growth factor-β (TGF-β), on the regulation of endothelin-1 (ET-1) mRNA and protein production in human amnion cells.
Methods:Human amnion cells were harvested from uncomplicated pregnancies undergoing elective cesarean delivery at term and grown in primary monolayer culture. Cells were treated with IL-1α, EGF, and TGF-β for dose-response and time course experiments. Northern analysis was used to determine ET-1 mRNA expression, and enzyme-linked immunosorbent assay was used for ET-1 peptide determination.
Results:Interleukin-1α, EGF, and TGF-β induced the expression of ET-1 mRNA and protein in a dose- and time-dependent fashion. The kinetics of ET-1 mRNA production did not differ markedly with respect to the inducing cytokine, but the kinetics of ET-1 protein production was quite different. Interleukin-1α and EGF stimulated a rapid increase in ET-1 that peaked by 24 hours, and the levels declined to just above the detection limit by 72 hours. In contrast, TGF-β-stimulated cells showed modest ET-1 production at early times (4-24 hours) and then gradually increased and peaked at 72 hours.
Conclusions:Cytokines modulate the expression of ET-1 mRNA and its cognate protein in human amnion cells. The differential kinetics of ET-1 peptide expression in amnion cells suggests that ET metabolism as well as synthesis contribute to the net expression of endothelin in amnion.
1. Yanagisawa M, Kurihara H, Tomobe Y, et al. A novel vasocontrictor peptide produced by vascular endothelial cells. Nature1988;332:411-415.
2.
2. Simonson MS, Dunn MJ. Endothelin peptides and the kidney. Ann Rev Physiol1993;55:249-265.
3.
3. Saito Y, Nakao K, Imura H. Endothelin in human disease. I. Essential hypertension, vasospastic angina, acute myocardial infarction, and chronic renal failure. In: Rubanyi GM, ed. Endothelin. New York: Oxford University Press, 1992:209-220.
4.
4. Stewart DJ. Endothelin in human disease. II. Shock, pulmonary hypertension, and congestive heart failure. In: Rubanyi GM, ed. Endothelin. New York: Oxford University Press, 1992:221-237.
5.
5. Inoue A, Yanagisawa M, Kimura S, et al. The human endothelin family: Three structurally and pharmacologicall distinct isopeptides predicted by three separate genes. Proc Natl Acad Sci USA1989;86:2863-2867.
6.
6. Nayer WG. Endothelin: isoforms, binding sites and possible implications in pathology. Trends Pharmacol Sci1990;11:96-99.
7.
7. Kamoi K, Sudo N, Ishibashi M, Yamaji T. Plasma endothelin-1 levels in patients with pregnancy-induced hypertension. N Engl J Med1990;323:1486-1487.
8.
8. Romero R, Avila C, Edwin SS, Mitchell MD. Endothelin-1,2 levels are elevated in the amniotic fluid of women with preterm labor and microbial invasion of the amniotic cavity. Am J Obstet Gynecol1992;166:95-99.
9.
9. Taylor RN, Varma M, Teng NN, Roberts JM. Women with preeclampsia have higher plasma endothelin levels than women with normal pregnancies. J Clin Endocrinol Metab1990;71:1675-1677.
11. Abassi ZA, Golomb E, Bridenbaugh R, Keiser HR. Metabolism of endothelin-1 and big endothelin by recombinant neutral endopeptidase 3.4.24.11. Br J Pharmacol1993;109:1024-1028.
12.
12. Devault A, Lazure C, Nault C, et al. Amino acid sequence of rabbit kidney neutral endopeptidase-24.11 (enkaphalinase) deduced from a complementary DNA. EMBO J1987;6:1317-1322.
13.
13. Matsas R, Fulcher IS, Kenny AJ, Turner AJ. Substance P and [leu]enkephalin are hydrolyzed by an enzyme in pig caudate synaptic membranes that is identical with the endopeptidase of kidney microvilli. Proc Natl Acad Sci USA1983;80:3111-3115.
14.
14. Howell S, Murray H, Turner AJ, Kenny AJ. A highly sensitive ELISA for endopeptidase-24.11, the common acute lymphoblastic leukaemia antigen (CALLA, CD-10), applicable to material of porcine and human origin. Biochem J1991;278:417-421.
15.
15. Kondepudi A, Johnson A. Cytokines increase neutral endopeptidase activity in lung fibroblasts. Am J Respir Cell Mol Biol1993;8:43-49.
16.
16. Casey ML, Smith JW, Nagal K, MacDonald PC. Transforming growth factor-β1 inhibits enkephalinase (EC 3.4.24.11) gene expression in human endometrial stromal cells and sex skin fibroblasts in culture. J Clin Endocrinol Metab1993;77:144-150.
17.
17. Turner AJ, Tanzawa K. Mammalian membrane metalloproteinases: NEP, ECE, KELL, and PEX, FASEB J1997;11:355-364.
18.
18. Friedman SA, Lubarsky SL, Ahokas RA, Nova A, Sibai BM. Preeclampsia and related disorders: Clinical aspects and relevance of endothelin and nitric oxide. Clin Perinatol1995;22:343-355.
19.
19. Casey ML, Word RA, MacDonald PC. Endothelin-1 gene expression and regulation of endothelin mRNA and protein biosynthesis in avascular human amnion: Potential source of amniotic fluid endothelin. J Biol Chem1991;266:5762-5768.
20.
20. Okita JR, Sagawa N, Casey ML, Snyder JM. A comparison of human amnion tissue and amnion cells in primary culture by morphological and biochemical criteria. In Vitro1983;19:117-126.
21.
21. Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein using the principle of protein dye binding. Anal Biochem1976;72:248-254.
22.
22. Chomczynski P, Sacchi N. Single step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem1987;162:156-159.
23.
23. Yoshizumi M, Kurihara H, Morita T, et al. Interleukin 1 increases the production of endothelin-1 by cultured endothelial cells. Biochem Biophys Res Comm1990;166:324-329.
24.
24. Kurihara H, Yoshizumi M, Sugiyama T, et al. Transforming growth factor-β stimulates the expression of endothelin mRNA by vascular endothelial cells. Biochem Biophys Res Comm1989;159:1435-1440.
25.
25. Economos K, MacDonald PC, Casey ML. Endothelin-1 gene expression and protein biosynthesis in human endometrium: Potential modulator of endometrial blood flow. J Clin Endocrinol Metab1992;74:14-19.
26.
26. Sunnergren KP, Word RA, Sambrook JF, MacDonald PC, Casey ML. Expression and regulation of endothelin precursor mRNA in avascular human amnion. Mol Cell Endocrinol1990;68:R7-R14.
27.
27. Xu D, Emoto N, Giaid A, et al. ECE-1: A membrane-bound metalloprotease that catalyses the proteolytic activation of big endothelin-1. Cell1994;78:473-485.
28.
28. Vijayaraghavan J, Scicli AG, Carretero OA, Slaughter C, Moomaw C, Hersh LB. The hydrolysis of endothelins by neutral endopeptidase 24.11 (enkaphalinase). J Biol Chem1990;265:14150-14155.
29.
29. Casey ML, MacDonald PC. The endothelin-parathyroid hormone-related protein vasoactive peptide system in human endometrium: modulation by transforming growth factor-β. Hum Reprod1996;11:62-82.
30.
30. Hasegawa M, Sagawa N, Itoh H, et al. Endothelin receptors in the human amnion, chorion laeve, decidua vera and placenta. Reprod Fertil Dev1995;7:1585-1589.
31.
31. Cameron IT, Davenport AP, Brown MJ, Smith SK. Endothelin-1 stimulates prostaglandin F2α release from human endometrium. Prostaglandins Leukot Essent Fatty Acids1991;42:155-157.
32.
32. Schrey MP, Hare A. Endothelin-1 stimulates phospholipid hydrolysis and prostaglandin F2α production in primary decidua cell cultures. Prostaglandins Leukot Essent Fatty Acids1992;47:321-325.
33.
33. Hughes AK, Padilla E, Kutchera WA, Michael JR, Kohan DE. Endothelin-1 induction of cyclooxygenase-2 expression in rat mesangial cells. Kidney Int1995;47:53-61.
34.
34. Kester M, Coroneos E, Thomas PJ, Dunn MJ. Endothelin stimulates prostaglandin endoperoxide synthase-2 mRNA expression and protein synthesis through a tyrosine kinase-signaling pathway in rat mesangial cells. J Biol Chem1994;269:22574-22580.
35.
35. Mitchell MD, Romero RJ, Lepera R, Rittenhouse L, Edwin SS. Actions of endothelin-1 on prostaglandin production by gestational tissues. Prostaglandins1990;40:627-635.
36.
36. Casey ML, Smith JW, Nagai K, Hersh LB, MacDonald PC. Progesterone-regulated cyclic modulation of membrane metalloendopeptidase (enkephalinase) in human endometrium. J Biol Chem1991;266:23041-23047.
37.
37. Wagner OF, Christ G, Wojta J, et al. polar secretion of endothelin-1 by cultured endothelial cells. J biol Chem1992;267:16066-16068.
38.
38. Romero R, Mazor M, Brandt F, et al. Interleukin-1α and interleukin-1β in preterm and term human parturition. Am J Reprod Immunol1992;27:117-123.
39.
39. Sagawa N. A study on the regulation of intrauterine milieu by bioactive substances of fetal origin in the amniotic fluid—the analyses of interactions among amnion, chorion laeve, and decidua vera tissues. Acta Obstet Gynaecol Jpn1994;46:686-696.
