- Email: [email protected]
Interleukin-1beta increases expression of tryptophan 2,3-dioxygenase and stimulates tryptophan metabolism in ectopic endometrial stromal cells
BMD significantly decreased in LD women from entry to both 24W (p<0.001) and 52W (p¼0.003). Hip BMD from entry also decreased in LD women compared to NA women at 52W (p<0.001). BMD for both groups recovered by 12FU when compared to entry (Lumbar: p¼0.054 and Hip: p¼0.903). BMD for NA use during 12FU had higher hip BMD at 12FU (p¼0.023). CONCLUSION: After one year of treatment with NA, BMD for women was not altered while lumbar spine BMD after LD then NA was significantly lower. This bone loss resolved over the next year. Women on NA were able to continue long-term treatment without experiencing a significant change in BMD. Supported by: NIH: R01-HD043281, Intramural Program, PRAE/NICHD and BCES/CC.
O-336 Wednesday, October 16, 2013 05:15 PM UNRAVELING THE MOLECULR INTERACTIONS BETWEEN PGE2 SIGNALING AND PAIN PATHWAYS IN ENDOMETRIOSIS. J. A. Arosh,a J. Lee,a M. Meagher,b K. Osteen,c K. Bruner-Tran,c S. Banu.a aIntegrative Bioscinecs, Texas A&M University, College Station, TX; bDepartment of Psychology, Texas A&M University, College Station, TX; cDepartment of Obstetrics and Gynecology, Vanderbilt University, Nashville, TN. OBJECTIVE: Prostaglandin E2 (PGE2) is the principal mediator in inflammation and pain hypersensitivity and plays important roles in the pathogenesis of endometriosis. PGE2 produced at the site of inflammation acts on the nociceptors of peripheral terminals through EP1, EP2, EP3, and EP4 receptors. Inhibition of PGE2 biosynthesis using NSAIDs and COX-2 inhibitors has emerged as the main class analgesics. Selective inhibition of PGE2 signaling down-stream of COX-2 may provide an opportunity to inhibit pronociceptive actions of PGE2 in endometriosis. DESIGN: Our working hypothesis is that pharmacological systemic blockade of EP2 and EP4 inhibits growth and innervations of endometriosis and peripheral and central nociceptive mechanisms. MATERIALS AND METHODS: Mixed population of human endometriotic epithelial cells (12Z-GFP) and stromal cells (22B-RFP) were xenografted into the peritoneal cavity Rag2g(c). On day 15 of xenograft, Group-1 (n¼6) mice were served as control. Group-2 (n¼6) mice were treated with EP2 and EP4 inhibitors. Group-3 (n¼6) were served as sham control. On day 30, motor activity of the experimental mice was examined by Digiscan. Then, the mice were euthanized and peritoneal endometriosis lesions were collected. RESULTS: A linear increase of 12Z-GFP or 22B-RFP cells was detected with the in vivo imager. Using a fluorescence stereo microscope, 18-20 lesions were detectable in the peritoneal cavity. Selective inhibition of EP2 and EP4 decreased growth of endometriosis about 50-60%, decreased expression of Protein Gene Product 9.5 (PGP 9.5), Substance P (SP), and Vesicular Monoamine Transporter (VMAT) in endometriosis innervations, and partially restored the motor activity in experimental endometriosis mice. CONCLUSION: These results together indicate that selective inhibition of EP2 and EP4 decreases growth of endometriosis, inhibits innervations of endometriosis and nociceptive pathways, and restores motor activity in endometriosis xenograft mice models. Supported by: 1R21HD066248-01A1 to JAA.
O-337 Wednesday, October 16, 2013 05:30 PM MULTIDETECTOR COMPUTERIZED TOMOGRAPHY ENTEROCLYSIS VERSUS MAGNETIC RESONANCE ENTEROCLYSIS IN THE DIAGNOSIS OF COLORECTAL ENDOMETRIOSIS. S. Ferrero,a U. Leone Roberti Maggiore,a P. L. Venturini,a G. A. Rollandi,b E. Biscaldi.b aDepartment of Obstetrics and Gynecology, San Martino Hospital and University of Genoa, Genoa, Liguria, Italy; bDepartment of Radiology, Galliera Hospital, Genoa, Liguria, Italy. OBJECTIVE: To compare the accuracy of multidetector computerized tomography enteroclysis (MDCT-e) and magnetic resonance enteroclysis
FERTILITY & STERILITYÒ
(MR-e) in determining the presence and the mucosa layer involvement of sigmoid and rectal endometriotic nodules. DESIGN: Single-center, prospective study. MATERIALS AND METHODS: Women with symptoms suggestive of bowel endometriosis underwent MDCT-e and MR-e. After retrograde colonic distension and injection of the intravenous contrast medium, patients were scanned on a 16-row MDCT scanner. MR-e was performed on a 1.5 T magnet using an 8 channels phased array coil; the intestinal distension was achieved by introducing in the rectum 250-300 ml of ultrasonographic gel diluted with saline solution. Radiological findings were compared with surgical and histological results. The McNemar’s test with the Yates continuity correction was used to compare the accuracy of the two techniques in the diagnosis of intestinal endometriosis. RESULTS: Two hundred and sixty patients were enrolled in the study; surgery demonstrated that 176 women (67.7%) had bowel endometriotic nodules. There was no difference in the accuracy of MDCT-e (98.5%) and MR-e (96.9%) in the diagnosis of sigmoid and rectal endometriosis (p ¼ 0.248). The sensitivity, specificity, positive predictive value, negative predictive value, positive likelihood ratio and negative likelihood ratio of MDCT-e and MR-e were respectively 98.3%, 98.8%, 99.4%, 96.5%, 81.59, 0.02 and 97.2%, 96.4%, 98.3%, 94.1%, 26.89, 0.03. MDCT-e accuracy had higher accuracy in determining the presence of endometriotic nodules infiltrating the mucosa layer compared with MR-e (p ¼ 0.041). CONCLUSION: Both MDCT-e and MR-e are accurate in the diagnosis of colorectal endometriosis. MDCT-e is more accurate than MR-e in determining correctly the depth of endometriotic nodule penetration in the intestinal wall.
O-338 Wednesday, October 16, 2013 05:45 PM INTERLEUKIN-1BETA INCREASES EXPRESSION OF TRYPTOPHAN 2,3-DIOXYGENASE AND STIMULATES TRYPTOPHAN METABOLISM IN ECTOPIC ENDOMETRIAL STROMAL CELLS. Y. Urata, Y. Osuga, K. Koga, Y. Hirota, T. Hirata, T. Fujii. Department of Obstetrics and Gynecology, University of Tokyo, Bunkyoku, Tokyo, Japan. OBJECTIVE: Tryptophan 2,3-dioxygenase (TDO) and indoleamine 2,3dioxygenase (IDO) are enzymes that catalyze the first and rate-limiting step of tryptophan degradation (i.e. from tryptophan to kynurenine) and thereby regulate tryptophan levels. Decrease in tryptophan levels inhibits T cell function and contributes to immune tolerance. While IFNg increases IDO expression, a molecule that regulates TDO expression remains to be elucidated. We evaluated the effect of interleukin (IL)-1b, a typical endometriosis-associated cytokine, on TDO expression in ectopic endometrial stromal cells (ESCs). DESIGN: Experimental study. MATERIALS AND METHODS: Under the approval of Institutional Review Board, ovarian endometriomas were obtained from women who had regular menstrual cycles and underwent laparoscopic surgery. ESCs were isolated from ovarian endometriomas. ESCs were treated with IL-1b for 5 hours to examine gene expression, for 24 hours to examine protein expression, and for 48 hours to examine the enzyme activity. Expression levels of TDO mRNA and IDO mRNA in ESCs were determined by quantitative real-time polymerase chain reaction. TDO protein was detected using western blotting. To study the enzyme activity of TDO and IDO, kynurenine concentration in the conditioned medium was measured. RESULTS: IL-1b (1 ng/ml) increased TDO mRNA expression to 6.3-fold of control but had no effect on IDO mRNA expression in ESCs. IL-1b (1 ng/ ml) increased TDO protein to 1.3-fold in ESCs and also increased kynurenine production to 1.9-fold in the conditioned medium. TDO siRNA suppressed the increase in IL-1b-induced kynurenine production. CONCLUSION: IL-1b increased TDO expression and stimulated tryptophan metabolism in ESCs. The IL-1b-induced increase in TDO expression may enhance immune tolerance to promote endometriosis. Supported by: Grants from the Ministry of Health, Labor, and Welfare and the Ministry of Education, Culture, Sports, Science, and Technology of Japan.
S103
O-336 Wednesday, October 16, 2013 05:15 PM UNRAVELING THE MOLECULR INTERACTIONS BETWEEN PGE2 SIGNALING AND PAIN PATHWAYS IN ENDOMETRIOSIS. J. A. Arosh,a J. Lee,a M. Meagher,b K. Osteen,c K. Bruner-Tran,c S. Banu.a aIntegrative Bioscinecs, Texas A&M University, College Station, TX; bDepartment of Psychology, Texas A&M University, College Station, TX; cDepartment of Obstetrics and Gynecology, Vanderbilt University, Nashville, TN. OBJECTIVE: Prostaglandin E2 (PGE2) is the principal mediator in inflammation and pain hypersensitivity and plays important roles in the pathogenesis of endometriosis. PGE2 produced at the site of inflammation acts on the nociceptors of peripheral terminals through EP1, EP2, EP3, and EP4 receptors. Inhibition of PGE2 biosynthesis using NSAIDs and COX-2 inhibitors has emerged as the main class analgesics. Selective inhibition of PGE2 signaling down-stream of COX-2 may provide an opportunity to inhibit pronociceptive actions of PGE2 in endometriosis. DESIGN: Our working hypothesis is that pharmacological systemic blockade of EP2 and EP4 inhibits growth and innervations of endometriosis and peripheral and central nociceptive mechanisms. MATERIALS AND METHODS: Mixed population of human endometriotic epithelial cells (12Z-GFP) and stromal cells (22B-RFP) were xenografted into the peritoneal cavity Rag2g(c). On day 15 of xenograft, Group-1 (n¼6) mice were served as control. Group-2 (n¼6) mice were treated with EP2 and EP4 inhibitors. Group-3 (n¼6) were served as sham control. On day 30, motor activity of the experimental mice was examined by Digiscan. Then, the mice were euthanized and peritoneal endometriosis lesions were collected. RESULTS: A linear increase of 12Z-GFP or 22B-RFP cells was detected with the in vivo imager. Using a fluorescence stereo microscope, 18-20 lesions were detectable in the peritoneal cavity. Selective inhibition of EP2 and EP4 decreased growth of endometriosis about 50-60%, decreased expression of Protein Gene Product 9.5 (PGP 9.5), Substance P (SP), and Vesicular Monoamine Transporter (VMAT) in endometriosis innervations, and partially restored the motor activity in experimental endometriosis mice. CONCLUSION: These results together indicate that selective inhibition of EP2 and EP4 decreases growth of endometriosis, inhibits innervations of endometriosis and nociceptive pathways, and restores motor activity in endometriosis xenograft mice models. Supported by: 1R21HD066248-01A1 to JAA.
O-337 Wednesday, October 16, 2013 05:30 PM MULTIDETECTOR COMPUTERIZED TOMOGRAPHY ENTEROCLYSIS VERSUS MAGNETIC RESONANCE ENTEROCLYSIS IN THE DIAGNOSIS OF COLORECTAL ENDOMETRIOSIS. S. Ferrero,a U. Leone Roberti Maggiore,a P. L. Venturini,a G. A. Rollandi,b E. Biscaldi.b aDepartment of Obstetrics and Gynecology, San Martino Hospital and University of Genoa, Genoa, Liguria, Italy; bDepartment of Radiology, Galliera Hospital, Genoa, Liguria, Italy. OBJECTIVE: To compare the accuracy of multidetector computerized tomography enteroclysis (MDCT-e) and magnetic resonance enteroclysis
FERTILITY & STERILITYÒ
(MR-e) in determining the presence and the mucosa layer involvement of sigmoid and rectal endometriotic nodules. DESIGN: Single-center, prospective study. MATERIALS AND METHODS: Women with symptoms suggestive of bowel endometriosis underwent MDCT-e and MR-e. After retrograde colonic distension and injection of the intravenous contrast medium, patients were scanned on a 16-row MDCT scanner. MR-e was performed on a 1.5 T magnet using an 8 channels phased array coil; the intestinal distension was achieved by introducing in the rectum 250-300 ml of ultrasonographic gel diluted with saline solution. Radiological findings were compared with surgical and histological results. The McNemar’s test with the Yates continuity correction was used to compare the accuracy of the two techniques in the diagnosis of intestinal endometriosis. RESULTS: Two hundred and sixty patients were enrolled in the study; surgery demonstrated that 176 women (67.7%) had bowel endometriotic nodules. There was no difference in the accuracy of MDCT-e (98.5%) and MR-e (96.9%) in the diagnosis of sigmoid and rectal endometriosis (p ¼ 0.248). The sensitivity, specificity, positive predictive value, negative predictive value, positive likelihood ratio and negative likelihood ratio of MDCT-e and MR-e were respectively 98.3%, 98.8%, 99.4%, 96.5%, 81.59, 0.02 and 97.2%, 96.4%, 98.3%, 94.1%, 26.89, 0.03. MDCT-e accuracy had higher accuracy in determining the presence of endometriotic nodules infiltrating the mucosa layer compared with MR-e (p ¼ 0.041). CONCLUSION: Both MDCT-e and MR-e are accurate in the diagnosis of colorectal endometriosis. MDCT-e is more accurate than MR-e in determining correctly the depth of endometriotic nodule penetration in the intestinal wall.
O-338 Wednesday, October 16, 2013 05:45 PM INTERLEUKIN-1BETA INCREASES EXPRESSION OF TRYPTOPHAN 2,3-DIOXYGENASE AND STIMULATES TRYPTOPHAN METABOLISM IN ECTOPIC ENDOMETRIAL STROMAL CELLS. Y. Urata, Y. Osuga, K. Koga, Y. Hirota, T. Hirata, T. Fujii. Department of Obstetrics and Gynecology, University of Tokyo, Bunkyoku, Tokyo, Japan. OBJECTIVE: Tryptophan 2,3-dioxygenase (TDO) and indoleamine 2,3dioxygenase (IDO) are enzymes that catalyze the first and rate-limiting step of tryptophan degradation (i.e. from tryptophan to kynurenine) and thereby regulate tryptophan levels. Decrease in tryptophan levels inhibits T cell function and contributes to immune tolerance. While IFNg increases IDO expression, a molecule that regulates TDO expression remains to be elucidated. We evaluated the effect of interleukin (IL)-1b, a typical endometriosis-associated cytokine, on TDO expression in ectopic endometrial stromal cells (ESCs). DESIGN: Experimental study. MATERIALS AND METHODS: Under the approval of Institutional Review Board, ovarian endometriomas were obtained from women who had regular menstrual cycles and underwent laparoscopic surgery. ESCs were isolated from ovarian endometriomas. ESCs were treated with IL-1b for 5 hours to examine gene expression, for 24 hours to examine protein expression, and for 48 hours to examine the enzyme activity. Expression levels of TDO mRNA and IDO mRNA in ESCs were determined by quantitative real-time polymerase chain reaction. TDO protein was detected using western blotting. To study the enzyme activity of TDO and IDO, kynurenine concentration in the conditioned medium was measured. RESULTS: IL-1b (1 ng/ml) increased TDO mRNA expression to 6.3-fold of control but had no effect on IDO mRNA expression in ESCs. IL-1b (1 ng/ ml) increased TDO protein to 1.3-fold in ESCs and also increased kynurenine production to 1.9-fold in the conditioned medium. TDO siRNA suppressed the increase in IL-1b-induced kynurenine production. CONCLUSION: IL-1b increased TDO expression and stimulated tryptophan metabolism in ESCs. The IL-1b-induced increase in TDO expression may enhance immune tolerance to promote endometriosis. Supported by: Grants from the Ministry of Health, Labor, and Welfare and the Ministry of Education, Culture, Sports, Science, and Technology of Japan.
S103