當子宮內膜的腺體上皮細胞與基質細胞在人體子宮腔外的任何其他部位附著增生時，即稱之為子宮內膜異位症。子宮內膜異位症是一種十分常見的婦科疾病，在具有生育年齡的婦女中約有10％ 罹患此疾病。許多證據都顯示出前列腺素E2和子宮內膜異位症的生長有很高的相關性。前列腺素E2是由一連串的酵素作用由磷脂質轉變而來，其中的速率決定步驟是由前列腺素G/H合成?所調控。本論文主要探討在異位的子宮內膜基質細胞上，介白素-1b透過何種機制去刺激前列腺素G/H合成?-2的表現以及其中所透過的訊息傳遞路徑。由實驗結果得知，前列腺素G/H合成?-2在異位的子宮內膜組織中有大量表現的情況。同時發現一個十分有趣的現象，和正常的子宮內膜基質細胞比較起來，異位的子宮內膜基質細胞對於介白素-1b的刺激更為敏感，其敏感度可達100倍之高。將正常或異位的子宮內膜基質細胞給予訊息傳遞的抑制劑BAY 11-7082(IKK選擇性抑制劑)，SB 202190(p38 MAPK 選擇性抑制劑) 以及 U 0126(MEK 選擇性抑制劑)，皆抑制了由介白素-1b所刺激的前列腺素G/H合成?-2的表現，因此推論介白素-1b能夠透過ERK1/2，p38，NF-kB的訊息路徑影響前列腺素G/H合成?-2的表現。利用反轉錄聚合?連鎖反應與西方轉漬法得知介白素-1第一型，第二型接受器以及介白素-1接受器的拮抗劑的表現在正常與異位子宮內膜基質細胞中無明顯差異。即時聚合?連鎖反應結果證實在正常與異位的子宮內膜基質細胞中，介白素-1b都能夠去穩定前列腺素G/H合成?-2傳訊者RNA的表現，並且同樣都是透過ERK1/2，p38與NF-kB的訊息傳遞路徑。利用細胞轉殖的方式送入前列腺素G/H合成?-2驅動子到正常或異位子宮內膜基質細胞中，螢光酵素檢測的結果得知在異位子宮內膜基質細胞中，介白素-1b能刺激前列腺素G/H合成?-2驅動子活性二至四倍增加，而在正常子宮內膜基質細胞中則完全無反應。利用序列刪除找出在前列腺素G/H合成?-2驅動子上的-571至-564之間的CRE site對於介白素-1b所刺激的驅動子活性是很重要的，並且推測就是透過CREB這個轉錄因子的調控作用。總而言之，在異位的子宮內膜基質細胞中，介白素-1b不但能夠穩定前列腺素G/H合成?-2傳訊者RNA，還能夠促進前列腺素G/H合成?-2驅動子的轉錄作用，這樣一個結果造成大量前列腺素的產生，因此促進了子宮內膜異位症的生成。

　　Endometriosis is defined as the presence of endometrial tissues within the pelvic peritoneum and/or other sites outsides-uterine cavity. It is the most commonly encountered gynecologic diseases affecting 10% of women in the reproductive age. Prostaglandin (PG) E2 has been found to be a key mediator in the development of endometriosis and its production was controlled by the first committed enzyme known as cyclooxygenase (COX). In this study, we attempted to determine the effect of interleukin-1b on COX-2 expression and the signal transduction pathways involved in endometriotic stromal cells. We found that COX-2 expression elevated in ectopic lesions compared with that in eutopic endometrium. IL-1b-mediated COX-2 expression and PGE2 production in endometriotic stromal cells was inhibited by NS-398, a selective COX-2 inhibitor. Interestingly, it was observed that COX-2 expression was 100 times more sensitive to IL-1b in endometriotic stromal cell than in normal endometrial stromal cells. Treatment with BAY 11-7082 (a selective inhibitor of IKK), SB 202190 (a selective cell-permeable p38 MAPK inhibitor) and U 0126 (MEK inhibitor) inhibited IL-1b-induced COX-2 expression in both stromal cells derived from endometriotic implant and normal endometrium. These results suggest NF-kB and MAPK signaling pathways were involved in IL1-b signalling in normal and endometriotic stromal cells. The results of RT- PCR and Western blot showed that there was no difference in IL-1b receptors (IL-1R1 and IL-1R2) expression between endometriotic and normal stromal cells. In addition, IL1-b enhanecd COX-2 mRNA stability through ERK1/2, p38 and NF-kB signal pathways in both normal and endometriotic stromal cells. In transient transfection assays, COX-2 promoter activity increased 2 to 4 fold after treatment with IL-1b in ectopic stromal cells while COX-2 promoter was unresponsive to IL-1b in normal stromal cells. By transfection with various COX-2 promoter constructs, we found that the -571 to -564 region was essential for IL-1b-induced COX-2 gene transcription and CRE element is present within this region. In conclusion, IL-1b stimulated COX-2 hyper-activation in endometriotic stromal cells with abnormal PGs production may contribute to the development and progression of endometriosis.

1. Kjerulff KH, Erickson BA, Langenberg PW 1996 Chronic gynecological conditions reported by US women: findings from the National Health Interview Survey, 1984 to 1992. Am J Public Health 86:195-9
2. Valle RF 2002 Endometriosis: current concepts and therapy. Int J Gynaecol Obstet 78:107-19
3. Jubanyik KJ, Comite F 1997 Extrapelvic endometriosis. Obstet Gynecol Clin North Am 24:411-40
4. Fernandez Pelegrina R, Kessler AG, Rawlins RG 1991 Modern approaches to the treatment of human infertility through assisted reproduction. P R Health Sci J 10:75-81
5. Candiani GB, Vercellini P, Fedele L, Colombo A, Candiani M 1991 Mild endometriosis and infertility: a critical review of epidemiologic data, diagnostic pitfalls, and classification limits. Obstet Gynecol Surv 46:374-82
6. Vinatier D, Orazi G, Cosson M, Dufour P 2001 Theories of endometriosis. Eur J Obstet Gynecol Reprod Biol 96:21-34
7. Leyendecker G, Herbertz M, Kunz G, Mall G 2002 Endometriosis results from the dislocation of basal endometrium. Hum Reprod 17:2725-36
8. Olive DL, Schwartz LB 1993 Endometriosis. N Engl J Med 328:1759-69
9. Gleicher N 1994 The role of humoral immunity in endometriosis. Acta Obstet Gynecol Scand Suppl 159:15-7
10. Dmowski WP, Steele RW, Baker GF 1981 Deficient cellular immunity in endometriosis. Am J Obstet Gynecol 141:377-83
11. Oosterlynck DJ, Cornillie FJ, Waer M, Vandeputte M, Koninckx PR 1991 Women with endometriosis show a defect in natural killer activity resulting in a decreased cytotoxicity to autologous endometrium. Fertil Steril 56:45-51
12. Halme J, Becker S, Hammond MG, Raj MH, Raj S 1983 Increased activation of pelvic macrophages in infertile women with mild endometriosis. Am J Obstet Gynecol 145:333-7
13. Vercellini P, De Benedetti F, Rossi E, Colombo A, Trespidi L, Crosignani PG 1993 Tumor necrosis factor in plasma and peritoneal fluid of women with and without endometriosis. Gynecol Obstet Invest 36:39-41
14. Cheong YC, Shelton JB, Laird SM, et al. 2002 IL-1, IL-6 and TNF-alpha concentrations in the peritoneal fluid of women with pelvic adhesions. Hum Reprod 17:69-75
15. Keenan JA, Chen TT, Chadwell NL, Torry DS, Caudle MR 1995 IL-1 beta, TNF-alpha, and IL-2 in peritoneal fluid and macrophage-conditioned media of women with endometriosis. Am J Reprod Immunol 34:381-5
16. Fukaya T, Sugawara J, Yoshida H, Yajima A 1994 The role of macrophage colony stimulating factor in the peritoneal fluid in infertile patients with endometriosis. Tohoku J Exp Med 172:221-6
17. Wu MY, Ho HN 2003 The role of cytokines in endometriosis. Am J Reprod Immunol 49:285-96
18. Halme J, White C, Kauma S, Estes J, Haskill S 1988 Peritoneal macrophages from patients with endometriosis release growth factor activity in vitro. J Clin Endocrinol Metab 66:1044-9
19. Raiter-Tenenbaum A, Baranao RI, Etchepareborda JJ, Meresman GF, Rumi LS 1998 Functional and phenotypic alterations in peritoneal macrophages from patients with early and advanced endometriosis. Arch Gynecol Obstet 261:147-57
20. Dinarello CA 1994 The interleukin-1 family: 10 years of discovery. Faseb J 8:1314-25
21. Gerard N, Caillaud M, Martoriati A, Goudet G, Lalmanach AC 2004 The interleukin-1 system and female reproduction. J Endocrinol 180:203-12
22. Loppnow H, Werdan K, Reuter G, Flad HD 1998 The interleukin-1 and interleukin-1 converting enzyme families in the cardiovascular system. Eur Cytokine Netw 9:675-80
23. Simon C, Piquette GN, Frances A, Polan ML 1993 Localization of interleukin-1 type I receptor and interleukin-1 beta in human endometrium throughout the menstrual cycle. J Clin Endocrinol Metab 77:549-55
24. Simon C, Piquette GN, Frances A, Westphal LM, Heinrichs WL, Polan ML 1993 Interleukin-1 type I receptor messenger ribonucleic acid expression in human endometrium throughout the menstrual cycle. Fertil Steril 59:791-6
25. Wu MH, Sun HS, Lin CC, et al. 2002 Distinct mechanisms regulate cyclooxygenase-1 and -2 in peritoneal macrophages of women with and without endometriosis. Mol Hum Reprod 8:1103-10
26. Kharfi A, Akoum A 2001 Correlation between decreased type-II interleukin-1 receptor and increased monocyte chemotactic protein-1 expression in the endometrium of women with endometriosis. Am J Reprod Immunol 45:193-9
27. Kharfi A, Boucher A, Akoum A 2002 Abnormal interleukin-1 receptor type II gene expression in the endometrium of women with endometriosis. Biol Reprod 66:401-6
28. Mori H, Sawairi M, Nakagawa M, Itoh N, Wada K, Tamaya T 1992 Expression of interleukin-1 (IL-1) beta messenger ribonucleic acid (mRNA) and IL-1 receptor antagonist mRNA in peritoneal macrophages from patients with endometriosis. Fertil Steril 57:535-42
29. Sahakian V, Anners J, Haskill S, Halme J 1993 Selective localization of interleukin-1 receptor antagonist in eutopic endometrium and endometriotic implants. Fertil Steril 60:276-9
30. Feng L, Sun W, Xia Y, et al. 1993 Cloning two isoforms of rat cyclooxygenase: differential regulation of their expression. Arch Biochem Biophys 307:361-8
31. O'Neill GP, Ford-Hutchinson AW 1993 Expression of mRNA for cyclooxygenase-1 and cyclooxygenase-2 in human tissues. FEBS Lett 330:156-60
32. Oral E, Arici A 1997 Pathogenesis of endometriosis. Obstet Gynecol Clin North Am 24:219-33
33. Williams CS, Mann M, DuBois RN 1999 The role of cyclooxygenases in inflammation, cancer, and development. Oncogene 18:7908-16
34. Chishima F, Hayakawa S, Sugita K, et al. 2002 Increased expression of cyclooxygenase-2 in local lesions of endometriosis patients. Am J Reprod Immunol 48:50-6
35. Fagotti A, Ferrandina G, Fanfani F, et al. 2004 Analysis of cyclooxygenase-2 (COX-2) expression in different sites of endometriosis and correlation with clinico-pathological parameters. Hum Reprod 19:393-7
36. Ota H, Igarashi S, Sasaki M, Tanaka T 2001 Distribution of cyclooxygenase-2 in eutopic and ectopic endometrium in endometriosis and adenomyosis. Hum Reprod 16:561-6
37. Tamura M, Sebastian S, Yang S, et al. 2002 pUp-regulation of cyclooxygenase-2 expression and rostaglandin synthesis in endometrial stromal cells by malignant endometrial epithelial cells. A paracrine effect mediated by prostaglandin E2 and nuclear factor-kappa B. J Biol Chem 277:26208-16
38. Noble LS, Takayama K, Zeitoun KM, et al. 1997 Prostaglandin E2 stimulates aromatase expression in endometriosis-derived stromal cells. J Clin Endocrinol Metab 82:600-6
39. Bulun SE, Yang S, Fang Z, et al. 2001 Role of aromatase in endometrial disease. J Steroid Biochem Mol Biol 79:19-25
40. Bulun SE, Zeitoun KM, Takayama K, Simpson E, Sasano H 2000 Aromatase as a therapeutic target in endometriosis. Trends Endocrinol Metab 11:22-7
41. Tsai SJ, Wu MH, Lin CC, Sun HS, Chen HM 2001 Regulation of steroidogenic acute regulatory protein expression and progesterone production in endometriotic stromal cells. J Clin Endocrinol Metab 86:5765-73
42. De Leon FD, Vijayakumar R, Rao CV, Yussman M 1988 Prostaglandin F2 alpha and E2 release by peritoneum with and without endometriosis. Int J Fertil 33:48-51
43. Karck U, Reister F, Schafer W, Zahradnik HP, Breckwoldt M 1996 PGE2 and PGF2 alpha release by human peritoneal macrophages in endometriosis. Prostaglandins 51:49-60
44. Johnson GL, Lapadat R 2002 Mitogen-activated protein kinase pathways mediated by ERK, JNK, and p38 protein kinases. Science 298:1911-2
45. Dunne A, O'Neill LA 2003 The interleukin-1 receptor/Toll-like receptor superfamily: signal transduction during inflammation and host defense. Sci STKE 2003:re3
46. Vanden Berghe W, De Bosscher K, Vermeulen L, De Wilde G, Haegeman G 2002 Induction and repression of NF-kappa B-driven inflammatory genes. Ernst Schering Res Found Workshop:233-78
47. Li Q, Verma IM 2002 NF-kappaB regulation in the immune system. Nat Rev Immunol 2:725-34
48. Li X, Stark GR 2002 NFkappaB-dependent signaling pathways. Exp Hematol 30:285-96
49. Poligone B, Baldwin AS 2001 Positive and negative regulation of NF-kappaB by COX-2: roles of different prostaglandins. J Biol Chem 276:38658-64
50. Tamura M, Sebastian S, Yang S, Gurates B, Fang Z, Bulun SE 2002 Interleukin-1beta elevates cyclooxygenase-2 protein level and enzyme activity via increasing its mRNA stability in human endometrial stromal cells: an effect mediated by extracellularly regulated kinases 1 and 2. J Clin Endocrinol Metab 87:3263-73
51. Jacobs AT, Ignarro LJ 2003 Nuclear factor-kappa B and mitogen-activated protein kinases mediate nitric oxide-enhanced transcriptional expression of interferon-beta. J Biol Chem 278:8018-27
52. Newton R, Kuitert LM, Bergmann M, Adcock IM, Barnes PJ 1997 Evidence for involvement of NF-kappaB in the transcriptional control of COX-2 gene expression by IL-1beta. Biochem Biophys Res Commun 237:28-32

53. Inoue H, Tanabe T 1998 Transcriptional role of the nuclear factor kappa B site in the induction by lipopolysaccharide and suppression by dexamethasone of cyclooxygenase-2 in U937 cells. Biochem Biophys Res Commun 244:143-8
54. Billack B, Heck DE, Mariano TM, et al. 2002 Induction of cyclooxygenase-2 by heat shock protein 60 in macrophages and endothelial cells. Am J Physiol Cell Physiol 283:C1267-77
55. Mestre JR, Rivadeneira DE, Mackrell PJ, et al. 2001 Overlapping CRE and E-box promoter elements can independently regulate COX-2 gene transcription in macrophages. FEBS Lett 496:147-51
56. Gorgoni B, Caivano M, Arizmendi C, Poli V 2001 The transcription factor C/EBPbeta is essential for inducible expression of the cox-2 gene in macrophages but not in fibroblasts. J Biol Chem 276:40769-77
57. Nie M, Pang L, Inoue H, Knox AJ 2003 Transcriptional regulation of cyclooxygenase 2 by bradykinin and interleukin-1beta in human airway smooth muscle cells: involvement of different promoter elements, transcription factors, and histone h4 acetylation. Mol Cell Biol 23:9233-44
58. Allport VC, Slater DM, Newton R, Bennett PR 2000 NF-kappaB and AP-1 are required for cyclo-oxygenase 2 gene expression in amnion epithelial cell line (WISH). Mol Hum Reprod 6:561-5
59. Joo M, Park GY, Wright JG, Blackwell TS, Atchison ML, Christman JW 2004 Transcriptional regulation of the cyclooxygenase-2 gene in macrophages by PU.1. J Biol Chem 279:6658-65
60. Sully G, Dean JL, Wait R, et al. 2004 Structural and functional dissection of a conserved destabilizing element of cyclo-oxygenase-2 mRNA: evidence against the involvement of AUF-1 [AU-rich element/poly(U)-binding/degradation factor-1], AUF-2, tristetraprolin, HuR (Hu antigen R) or FBP1 (far-upstream-sequence-element-binding protein 1). Biochem J 377:629-39
61. Witz CA 1999 Current concepts in the pathogenesis of endometriosis. Clin Obstet Gynecol 42:566-85
62. Kelly RW, King AE, Critchley HO 2001 Cytokine control in human endometrium. Reproduction 121:3-19
63. Lebovic DI, Bentzien F, Chao VA, Garrett EN, Meng YG, Taylor RN 2000 Induction of an angiogenic phenotype in endometriotic stromal cell cultures by interleukin-1beta. Mol Hum Reprod 6:269-75
64. Akoum A, Jolicoeur C, Kharfi A, Aube M 2001 Decreased expression of the decoy interleukin-1 receptor type II in human endometriosis. Am J Pathol 158:481-9
65. Dixon DA, Kaplan CD, McIntyre TM, Zimmerman GA, Prescott SM 2000 Post-transcriptional control of cyclooxygenase-2 gene expression. The role of the 3'-untranslated region. J Biol Chem 275:11750-7
66. Guhaniyogi J, Brewer G 2001 Regulation of mRNA stability in mammalian cells. Gene 265:11-23
67. Lasa M, Mahtani KR, Finch A, Brewer G, Saklatvala J, Clark AR 2000 Regulation of cyclooxygenase 2 mRNA stability by the mitogen-activated protein kinase p38 signaling cascade. Mol Cell Biol 20:4265-74
68. Subbaramaiah K, Marmo TP, Dixon DA, Dannenberg AJ 2003 Regulation of cyclooxgenase-2 mRNA stability by taxanes: evidence for involvement of p38, MAPKAPK-2, and HuR. J Biol Chem 278:37637-47
69. Dixon DA, Tolley ND, King PH, et al. 2001 Altered expression of the mRNA stability factor HuR promotes cyclooxygenase-2 expression in colon cancer cells. J Clin Invest 108:1657-65
70. Nabors LB, Gillespie GY, Harkins L, King PH 2001 HuR, a RNA stability factor, is expressed in malignant brain tumors and binds to adenine- and uridine-rich elements within the 3' untranslated regions of cytokine and angiogenic factor mRNAs. Cancer Res 61:2154-61
71. Caivano M, Gorgoni B, Cohen P, Poli V 2001 The induction of cyclooxygenase-2 mRNA in macrophages is biphasic and requires both CCAAT enhancer-binding protein beta (C/EBP beta ) and C/EBP delta transcription factors. J Biol Chem 276:48693-701
72. Eliopoulos AG, Dumitru CD, Wang CC, Cho J, Tsichlis PN 2002 Induction of COX-2 by LPS in macrophages is regulated by Tpl2-dependent CREB activation signals. Embo J 21:4831-40
73. Della Fazia MA, Servillo G, Sassone-Corsi P 1997 Cyclic AMP signalling and cellular proliferation: regulation of CREB and CREM. FEBS Lett 410:22-4