SAT-023 Cyclic Regulation of 11 Beta-Hydroxysteroid Dehydrogenase Type 1 in Endometriosis in Rhesus Macaques

Abstract

Endometriosis is an estrogen-dependent disorder defined by endometrium-like tissue outside the uterine cavity that results in pelvic inflammation. Pro-inflammatory cytokines can increase 11 β -hydroxysteroid dehydrogenase type 1 (11β-HSD1), which converts inactive cortisone into active cortisol. We hypothesized that cyclic changes in estrogen (estradiol; E2) and progesterone (P4) could regulate endometriotic 11β-HSD1. To test this, we assessed 11β-HSD1 expression in rhesus macaques (Macaca mulatta) with advanced endometriosis. Animal care was provided by our Department of Comparative Medicine and all work was reviewed and approved by the Institutional Animal Care and Use Committee. Endometrium and lesions were collected in the proliferative and secretory phase of the cycle (n=5 each). Control endometrium (endometriosis-free animals) was also collected in the proliferative (n=5) and secretory (n=4) phase. Mean (+SE) hormone levels were: 103+10 pg E2/ml and 0.2 + 0.04 ng P/ml in the proliferative phase; 43.8 + 9 pg E2/ml and 4.41 + 0.98 ng P/ml in the secretory phase. Tissues were frozen in liquid nitrogen for RNA isolation or fixed for histology. RNA was isolated in TRIzol and purified on Qiagen Quick columns with DNase digestion. Total RNA (1 µg) was reverse transcribed in the presence of random hexamer primers. TaqMan® quantitative real-time PCR (q-RT-PCR) was conducted for 11β-HSD1, estrogen receptor 1 (ESR1), ESR2 and progesterone receptor (PGR). Results are presented as ratio of target/ribosomal S10 RNA. Normal cyclic regulation of endometrial histology was observed. Histology confirmed the presence of endometrium-like tissue in all lesions. The highest expression of 11β-HSD1 was in proliferative phase endometriotic lesions (0.64 + 0.28). Levels were reduced in the secretory phase (0.13 + 0.02; P<0.01). 11β-HSD1 was minimal in the endometrium of control and endometriosis animals (proliferative phase control = 0.11 + 0.03; versus endometriotic animals = 0.11+ 0.06). There was a non-significant trend toward higher levels in the secretory phase (control = 0.24+ 0.06; endometriosis = 0.16 + 0.03). Normal cyclic levels of ESR1 and PGR were observed. For instance, levels of ESR1 (proliferative versus secretory) in the control endometrium, endometriosis endometrium, and lesions were as follows: 2.36 + 0.39 versus 0.43 + 0.21 (P<0.01), 1.41 + 0.22 versus 0.74 + 0.34 (P<0.01), and 1.05 + 0.35 versus 0.48 + 0.11(P<0.05); respectively. Levels of PGR followed the pattern of ESR1. In contrast, ESR2 transcript was higher in lesions compared to endometrium, and menstrual cycle-specific regulation was observed. We conclude that 11β-HSD1 is elevated in the proliferative phase resulting in increased intracellular cortisol. Elevated cortisol could promote estrogen dependent lesion growth by modulating inflammation. This work was supported by NIH/NICHD.