Interleukin-1 Stimulates Macrophage Migration Inhibitory Factor Secretion in Ectopic Endometrial Cells of Women With Endometriosis

SHBG or insulin concentrations were analyzed. 5␣-R1 mRNA and AR mRNA and protein were present in the intrauterine endometria of both groups, as well as in the pelvic implants of the endometriosis group. The same topographic distribution of AR and mRNA abundance for AR and 5␣-R1 were observed regardless of the study group or the tissue being eutopic or ectopic. CONCLUSIONS: Infertile women with endometriosis do not appear to be more exposed to circulating testosterone nor to androgen stimulation at the endometrium compared to other infertile women without evidence of endometriosis. Supported by: CNPq, Pronex P-133 Leuprolide Acetate and Hormonal Add-Back in Stage IV Endometriosis Patients With Chronic Pelvic Pain: A 10-Year Follow Up. M. A. Bedaiwy, R. F. Casper. Mount Sinai Hospital, Toronto, ON, Canada. OBJECTIVE: Treatment of symptomatic endometriosis with gonadotropin-releasing hormone-agonist (GnRH-agonist) down-regulation is typically limited to a 6 month period because of side effects related to hypoestrogenism, especially loss of bone mineral density (BMD). We and others have previously reported the use of replacement or “addback” of estrogen and/or progestin to the GnRH-agonist therapy to prevent hot flushes and bone loss, although the results regarding BMD have been mixed. The objective of this study is to examine the effect on BMD of a low-dose estrogen and pulsed progestogen hormone therapy (HT) regimen for addback during long-term GnRH-agonist therapy DESIGN: A pilot clinical trial. MATERIALS AND METHODS: The study included 5 symptomatic patients with stage IV endometriosis, who were aged - to - at the start of treatment. Patients received leuprolide acetate depot 3.75 mg IM monthly together with HT addback. The HT regimen consisted of 1 mg oral micronized estradiol daily continuously and 0.35 mg norethindrone daily for 2 days alternating with 2 days off norethindrone. The main outcome measure was bone density assessment in the lumbar spine, femoral neck and total hip region by dual-energy x-ray absorptiometry (DEXA) at yearly intervals. The mean follow-up duration (⫾ SD) while on GnRH-agonist treatment was 8.5 ⫾ 2.35 years. RESULTS: The mean age at the end of follow-up (⫾ SD) was 40.33⫾ 8.5 years. Bone mineral density was stable after initiation of HT for the entire follow-up period (Figure 1). One patient stopped her treatment on two occasions in order to conceive and was successful each time with delivery of a normal baby. She resumed treatment after each pregnancy. All other patients continued treatment for the duration of the study. No patient had return of pelvic pain after HT add-back.

CONCLUSIONS: Long-term GnRH-agonist down-regulation combined with low dose HT add-back appears to be effective in maintaining BMD and in providing symptomatic relief for patients with advanced endometriosis. Supported by: None

FERTILITY & STERILITY威

P-134 Interleukin-1 Stimulates Macrophage Migration Inhibitory Factor Secretion in Ectopic Endometrial Cells of Women With Endometriosis. C. Herrmann Lavoie, M. Therriault, R. Maheux, A. Akoum. Universite´ Laval, Que´bec, PQ, Canada. OBJECTIVE: To determine the effect of interleukin-1 (IL-1) on macrophage inhibitory factor (MIF) expression in endometriotic cells. DESIGN: Primary cultures of endometriotic cells isolated from human endometriotic lesions were exposed to different concentrations of IL-1. Expression of MIF was analyzed at the protein and the mRNA levels. MATERIALS AND METHODS: Endometriotic cells were isolated from endometriotic lesions and cultured according to routinely used procedures. Cells grown to confluence were incubated overnight with DMEM-F12 then stimulated with different concentrations of IL-1 (0 to 1 ng/mL) for different periods of time (0-24h). MIF secretion was evaluated by immunocytofluorescence and ELISA, whereas MIF mRNA levels were analyzed by RTPCR and Southern blotting. Electrophoretic mobility shift assay (EMSA) was used for determine whether IL-1 effects were mediated by the transcription factor NF␬B. RESULTS: Immunocytofluorescence showed that IL-1 increased MIF secretion in endometriotic cells. ELISA measurement of MIF secretion in the culture medium showed that IL-1 acts rapidly on endometriotic cells (30 minutes after stimulation) and exhibited a dose and time-dependent stimulatory effect. RT-PCR analysis showed an increase of MIF mRNA levels after IL-1 stimulation. Gelshift assays showed that IL-1 activates NF␬B and that curcumin, an NF␬B inhibitor, inhibited IL-1-mediated MIF secretion and NF␬B activation. CONCLUSIONS: The cytokine IL-1 acts on endometriotic cells by increasing MIF production. Such an interaction between IL-1 and MIF may have an important impact on endometriotic cell growth, considering the biological properties of the two major cytokines and their role in the pathophysiology of endometriosis. Supported by: This work was supported by grant MOP-37921 (to A.A.) from the Canadien Institutes for Health Research.

P-135 Procollagen Aminoterminal Propeptide Type I in Endometriosis. S. Ferrero, D. J. Gillott, V. Remorgida, N. Ragni, B. Teisner, J. G. Grudzinskas. San Martino Hospital, University of Genoa, Genoa, Italy; Reproductive Physiology Laboratory, St Bartholomew’s Hospital, QMW College, London, United Kingdom; Department of Medical Microbiology, University of Southern Denmark, Odense University, Odense, Denmark; The London Bridge Fertility, Gynaecology and Genetics Centre, London, United Kingdom. OBJECTIVE: Fibrosis is frequent in endometriotic lesions; procollagen aminoterminal propeptide type I (PINP) is a degradation product of collagen synthesis. This study aims to measure the levels of PINP in peritoneal fluid (PF) and plasma (PL) of women with and without endometriosis, in order to evaluate whether they correlate with the severity of the disease. DESIGN: Cross-sectional study. MATERIALS AND METHODS: PF and PL samples were obtained from 93 women with endometriosis and 68 controls (infertility, n⫽16; uterine myomas, n⫽25; ovarian cyst, n⫽21; pelvic pain, n⫽5; tubal sterilization, n⫽1). Only patients of reproductive age were included in the study; none of the subjects had used hormonal therapies in the 6 months before surgery. The extent of endometriosis was scored using the revised American Fertility Society (rAFS) classification. The phase of the cycle was determined according to the menstrual history and progesterone levels. The serum concentration of PINP was measured by a sandwich enzyme-linked immunosorbent assay utilizing purified alpha 1-chain specific rabbit antibodies. PINP concentration was not normally distributed as determined by the Shapiro-Wilks test; therefore, the Mann-Whitney U test was used to compare its concentration between the two groups. The relationship between PL and PF PINP concentration was evaluated by the non-parametric Spearman correlation coefficient. RESULTS: No significant difference was observed in the mean age of patients in the two groups. PINP concentration was significantly higher in PF in women with severe endometriosis (rAFS stage III-IV, mean ⫾ SEM, 1067.2⫾198.4 ng/mL) than in mild endometriosis (rAFS stage I-II,

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