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Sildenafil prevents apoptosis of first trimester trophoblast cells exposed to environmental stress
DESIGN: Longitudinal cohort study. MATERIALS AND METHODS: The study linked cycles from the Society for Assisted Reproductive Technology Clinic Outcomes Reporting System (SART CORS) for all ART clinics in Massachusetts between 2004 and 2009 with the resultant live births and fetal deaths. Odds ratios and 95% confidence intervals of adverse birth outcomes were calculated, controlling for maternal age, education, race/ethnicity, marital status, nativity, parity, payer, chronic diabetes and hypertension, with births to the fertile group as the reference group. RESULTS: The study population included a total of 340,517 births. Birth outcomes are shown in Table 1 for singletons and twins. Among singleton births, ART conceived births show higher rates of preterm birth and low birth weight, but not birth defects. Among twins, ART births were comparable to births to mothers with indicators of subfertility on all three outcomes. CONCLUSION: Outcomes of births conceived by ART did not differ from outcomes of births to mothers with indicators of subfertility in terms of birth defects and recorded higher rates of preterm birth and low birthweight only in singleton births. Supported by: NIH grants:R01HD067270; R01HD064595 & SART. O-167 Tuesday, October 15, 2013 12:15 PM EFFECT OF LETROZOLE VERSUS CLOMIPHENE ON LIVE BIRTH IN WOMEN WITH ANOVULATORY INFERTILITY DUE TO POLYCYSTIC OVARY SYNDROME (PCOS): A RANDOMIZED DOUBLE-BLIND MULTI-CENTER TRIAL. NIH/NICHD Reproductive Medicine Network. Fertility and Infertility Branch, Eunice Kennedy Shriver NICHD, Bethesda, MD. OBJECTIVE: To determine first line infertility therapy in PCOS. Clomiphene(CC) is the standard, but aromatase inhibitors may have more favorable endometrial effects and lower multiple pregnancy(MP) rates. DESIGN: RCT of 2 regimens(Letrozole(LTZ) vs CC) for up to 5 treatment cycles with monthly baseline and midluteal visits to determine ovulation(OV)/pregnancy(PREG) and tracking of PREG outcomes. Primary outcome: Live birth(LB). Secondary outcomes: OV, PREG loss(PL) and MP rates. MATERIALS AND METHODS: 750 PCOS women were randomized. PCOS was defined by Rotterdam Criteria: oligomenorrhea %8 menses/ yr plus either PCO on ultrasound or clinical/biochemical hyperandrogenism with exclusion of other causes. Females were 18-39y, had at least 1 patent fallopian tube and normal uterine cavity and male partner with sperm concentration of R 14 million/mL who consented to regular intercourse. RESULTS: The treatment groups were well matched for baseline historical, demographic and biochemical parameters[LTZ (N¼374) Age 29(5)y, BMI 35(10)] vs CC(N¼376)Age 28(4), BMI 35(9)[(mean(SD)]. Cumulative LB occurred more frequently in women treated with LTZ[27.5% vs CC:19.5%, P¼.007, Relative Risk of LB:1.44, 95%CI(1.10-1.87)] with no difference in birth weight between groups [(LTZ: 3232g(657) vs CC:3220g(715)]. The LB benefit was more in obese women(BMIR35) but LTZ was equal or superior to CC at all BMI groups with similar pregnancy benefit noted with LTZ in infertility treatment na€ıve(N ¼ 334) and CC na€ıve subjects(N ¼ 397). The OV rate was superior with LTZ(61.7% vs CC:48.3%,P<.0001). There was no significant difference in PL after conception (LTZ:31.8% vs CC:28.2%) or MP rates (all twins) (LTZ :3.2% vs CC:7.4%) between groups. The number of serious adverse events and congenital anomaly rate(with additional screening within 1 month of birth by trained pediatric personnel) were not different between groups. CONCLUSION: LTZ improves ovulation and live birth rates in infertile women with PCOS. We conclude that LTZ is preferable to CC as first line therapy. O-168 Tuesday, October 15, 2013 12:30 PM COMPARISON OF MONTELUKAST AND CABERGOLINE FOR PREVENTION OF OVARIAN HYPERSTIMULATION SYNDROME (OHSS): IN AN EXPERIMENTAL RAT MODEL. L. Akman,a G. Sahin,b O. Erbas,c H. Aktug,d A. Akdogan,b E. N. Tavmergen Goker.a,b a Obstetrics and Gynecology, Ege University Medical School, Izmir, Turkey; b Family Planning and Infertility Research and Treatment Center, Ege University, Izmir, Turkey; cPhysiology, Ege University Medical School, Izmir, Turkey; dHistology and Embrylogy, Ege University Medical School, Izmir, Turkey.
FERTILITY & STERILITYÒ
OBJECTIVE: To investigate the effects of the montelukast (leukotriene receptor antagonist) treatment in prevention of OHSS and compare to cabergoline treatment. DESIGN: 28 immature female Wistar rats were used for this randomized, controlled experimental study. The study groups were as following; the Group 1(control) received 0.1mL saline, Group 2(OHSS model);10 IU pregnant mare serum gonadotropin (PMSG)+30 IU hCG, Group 3(OHSS+Cb2); 10 IU PMSG+ 30 IU hCG+ 100 mg/kg Cb2 and Group 4 (OHSS+montelukast);10 IU PMSG+30 IU hCG+20 mg/kg montelukast. MATERIALS AND METHODS: PMSG was given intra-peritoneally for 4 consecutive days and HCG was given subcutaneously on the fifth day to induce OHSS. Treatment agents (Cb2-Dostinex and montelukast-Singulair) were given 2 hours before each PMSG injection during 4 days and the day of hCG. 48 hours after hCG administration, vascular permeability was evaluated by detection of injected Evans Blue in the peritoneal fluid. Rats were sacrificed after oophorectomy. Body weight, ovarian weight, VP and VEGF immune-expression were compared between the Cb2 and montelukast groups. RESULTS: Body weight, ovarian weight, VP and VEGF expression were significantly increased in the OHSS group compared to control. Both Cb2 and montelukast prevented progression of OHSS compared to OHSS group, significantly reduced body weight (58.23 6.45 gr and 61.34 4.86 gr vs 73.27 5.54 gr, p< 0.05, respectively), ovarian weight (26.46 2.12 mg and 28.84 3.45 mg vs 45.64 5.72, p<0.05, respectively), VP (19.5 0.18 mM/100 g and 23.75 0.63 mM/100 g vs 29.8 2.23 mM/100 g, p< 0.05, respectively) and VEGF expression (% 8.64 2.42 and % 10.75 1.61 vs % 20.18 3.45, p<0.05, respectively). CONCLUSION: Montelukast is an effective option for prevention of OHSS, as well as cabergoline. Montelukast may be a new treatment option to prevent and control OHSS. REPRODUCTIVE ENDOCRINOLOGY AND INFERTILITY FELLOWS RESEARCH II O-169 Tuesday, October 15, 2013 04:00 PM SILDENAFIL PREVENTS APOPTOSIS OF FIRST TRIMESTER TROPHOBLAST CELLS EXPOSED TO ENVIRONMENTAL STRESS. J. M. Bolnick,a B. A. Kilburn,a M. Singh,a M. P. Diamond,b M. Hertz,a D. R. Armant.a,c aOb/Gyn, Wayne State University, Detroit, MI; bOb/Gyn, Georgia Regents University, Athens, GA; cPRAE, NICHD, NIH, Bethesda, MD. OBJECTIVE: Human trophoblast cells develop within a hypoxic environment during the first trimester of pregnancy. Oxygen fluctuations can create environmental stress and induce trophoblast apoptosis, leading to placental dysfunction. Sildenafil (Viagra) is a potent, selective inhibitor of phosphodiesterase type 5, which increases guanosine 3’,5’-cyclic monophosphate (cGMP). We evaluated whether Sildenafil could rescue human trophoblasts from apoptosis caused by ischemic reperfusion injury using first trimester chorionic villous explants and the cytotrophoblast cell line, HTR-8/SVneo. DESIGN: In vitro study of placental tissue discarded from patients undergoing elective terminations during the first trimester and HTR-8/SVneo cells. MATERIALS AND METHODS: Villous explants or HTR-8/SVneo were treated with Sildenafil with or without a specific cGMP agonist, cGMP antagonist, NO agonist SNAP, or NO antagonist L-NAME. Treatments were performed by adding 350 ng/ml Sildenafil at 2% oxygen, 20% oxygen or with exposure to hypoxia/reoxygenation (H/R), defined as 2 h at 2% oxygen and 6 h at 20% oxygen. Cell death was evaluated using the TUNEL method and VEGF was determined by immunohistochemistry. RESULTS: Treatment with H/R significantly increased cell death when compared with villi or cells cultured continuously at 2% or 20% oxygen. Restoration of cell survival was achieved by adding Sildenafil during reoxygenation, but not in the presence of antagonists of cGMP or NO. Conversely, cGMP agonist and SNAP were cytoprotective. Sildenafil significantly increased VEGF. CONCLUSION: Trophoblast exposure to fluctuating oxygen hinders survival, which could precipitate significant adverse pregnancy outcomes. Sildenafil reduces apoptosis by elevation of cGMP, which is downstream of NO. This pathway can possibly prevent placental insufficiency. Sildenafil may also play a pivotal role in implantation through it’s angiogenic properties in upregulating VEGF. Supported by: Wayne State University and the Intramural Research Program of the NICHD, NIH.
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FERTILITY & STERILITYÒ
OBJECTIVE: To investigate the effects of the montelukast (leukotriene receptor antagonist) treatment in prevention of OHSS and compare to cabergoline treatment. DESIGN: 28 immature female Wistar rats were used for this randomized, controlled experimental study. The study groups were as following; the Group 1(control) received 0.1mL saline, Group 2(OHSS model);10 IU pregnant mare serum gonadotropin (PMSG)+30 IU hCG, Group 3(OHSS+Cb2); 10 IU PMSG+ 30 IU hCG+ 100 mg/kg Cb2 and Group 4 (OHSS+montelukast);10 IU PMSG+30 IU hCG+20 mg/kg montelukast. MATERIALS AND METHODS: PMSG was given intra-peritoneally for 4 consecutive days and HCG was given subcutaneously on the fifth day to induce OHSS. Treatment agents (Cb2-Dostinex and montelukast-Singulair) were given 2 hours before each PMSG injection during 4 days and the day of hCG. 48 hours after hCG administration, vascular permeability was evaluated by detection of injected Evans Blue in the peritoneal fluid. Rats were sacrificed after oophorectomy. Body weight, ovarian weight, VP and VEGF immune-expression were compared between the Cb2 and montelukast groups. RESULTS: Body weight, ovarian weight, VP and VEGF expression were significantly increased in the OHSS group compared to control. Both Cb2 and montelukast prevented progression of OHSS compared to OHSS group, significantly reduced body weight (58.23 6.45 gr and 61.34 4.86 gr vs 73.27 5.54 gr, p< 0.05, respectively), ovarian weight (26.46 2.12 mg and 28.84 3.45 mg vs 45.64 5.72, p<0.05, respectively), VP (19.5 0.18 mM/100 g and 23.75 0.63 mM/100 g vs 29.8 2.23 mM/100 g, p< 0.05, respectively) and VEGF expression (% 8.64 2.42 and % 10.75 1.61 vs % 20.18 3.45, p<0.05, respectively). CONCLUSION: Montelukast is an effective option for prevention of OHSS, as well as cabergoline. Montelukast may be a new treatment option to prevent and control OHSS. REPRODUCTIVE ENDOCRINOLOGY AND INFERTILITY FELLOWS RESEARCH II O-169 Tuesday, October 15, 2013 04:00 PM SILDENAFIL PREVENTS APOPTOSIS OF FIRST TRIMESTER TROPHOBLAST CELLS EXPOSED TO ENVIRONMENTAL STRESS. J. M. Bolnick,a B. A. Kilburn,a M. Singh,a M. P. Diamond,b M. Hertz,a D. R. Armant.a,c aOb/Gyn, Wayne State University, Detroit, MI; bOb/Gyn, Georgia Regents University, Athens, GA; cPRAE, NICHD, NIH, Bethesda, MD. OBJECTIVE: Human trophoblast cells develop within a hypoxic environment during the first trimester of pregnancy. Oxygen fluctuations can create environmental stress and induce trophoblast apoptosis, leading to placental dysfunction. Sildenafil (Viagra) is a potent, selective inhibitor of phosphodiesterase type 5, which increases guanosine 3’,5’-cyclic monophosphate (cGMP). We evaluated whether Sildenafil could rescue human trophoblasts from apoptosis caused by ischemic reperfusion injury using first trimester chorionic villous explants and the cytotrophoblast cell line, HTR-8/SVneo. DESIGN: In vitro study of placental tissue discarded from patients undergoing elective terminations during the first trimester and HTR-8/SVneo cells. MATERIALS AND METHODS: Villous explants or HTR-8/SVneo were treated with Sildenafil with or without a specific cGMP agonist, cGMP antagonist, NO agonist SNAP, or NO antagonist L-NAME. Treatments were performed by adding 350 ng/ml Sildenafil at 2% oxygen, 20% oxygen or with exposure to hypoxia/reoxygenation (H/R), defined as 2 h at 2% oxygen and 6 h at 20% oxygen. Cell death was evaluated using the TUNEL method and VEGF was determined by immunohistochemistry. RESULTS: Treatment with H/R significantly increased cell death when compared with villi or cells cultured continuously at 2% or 20% oxygen. Restoration of cell survival was achieved by adding Sildenafil during reoxygenation, but not in the presence of antagonists of cGMP or NO. Conversely, cGMP agonist and SNAP were cytoprotective. Sildenafil significantly increased VEGF. CONCLUSION: Trophoblast exposure to fluctuating oxygen hinders survival, which could precipitate significant adverse pregnancy outcomes. Sildenafil reduces apoptosis by elevation of cGMP, which is downstream of NO. This pathway can possibly prevent placental insufficiency. Sildenafil may also play a pivotal role in implantation through it’s angiogenic properties in upregulating VEGF. Supported by: Wayne State University and the Intramural Research Program of the NICHD, NIH.
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