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Endothelial nitric oxide synthase in uteroplacental vasculature in an ovine model of IUGR
SMFM Abstracts S193
Volume 189, Number 6 Am J Obstet Gynecol 487
ENDOTHELIAL NITRIC OXIDE SYNTHASE IN UTEROPLACENTAL VASCULATURE IN AN OVINE MODEL OF IUGR ALESSANDRA PADOAN1, TIMOTHY R.H. REGNAULT2, BARBRA DEVRIJER2, SEAN W. LIMESAND2, RUSSELL V. ANTHONY2, ENRICO FERRAZZI1, RANDALL B. WILKENING2, HENRY L. GALAN3, 1University of Milan, Milan, Italy 2University of Colorado Health Sciences Center, Pediatrics, aurora, CO 3University of Colorado Health Sciences Center, Obstetrics and Gynecology, Denver, CO OBJECTIVE: Endothelial nitric oxide synthase (eNOS) modulates blood flow and is upregulated by hypoxia. Hypothesis: eNOS is upregulated in the uterine artery (UtA), umbilical artery (UmA) and umbilical vein (UmV) in an ovine model of IUGR. STUDY DESIGN: 6 ewes were exposed to hyperthermic conditions from 35115 dGA to induce IUGR and then placed with 6 control ewes in ambient conditions. At 125dGA UmA Doppler pulsatility indices (PI) were assessed and catheters placed for umbilical blood flow (UF) and arterial gas measurements prior to sacrifice at 135dGA. UmA, UmV and UtA were prepared for eNOS Western blot analysis. Data (mean ± SE) were analyzed with t-tests and regression analysis. RESULTS: IUGR birthweights were reduced compared to controls (3.1 ± 1.8kg v 1.5 ± 1.6kg; p < 0.01). See table for results. No differences were seen for pCO2 and pH. UmA PI and UmA [eNOS} correlated well (R = 0.6). CONCLUSION: eNOS protein is largely increased in all vessels, which may be due to a hypoxic uteroplacental environment. Hypoxia, increased placental vascular resistance (UmA PI) and thus shear stress, may account for eNOS upregulation in the UmA and it’s placental branches.(supported by WRHR grant).
489
BIRTHWEIGHT AS A FUNCTION OF MATERNAL DUTY STATUS AND PARTICIPATION IN MANDATORY PHYSICAL TRAINING PROGRAM RODERICK HUME1, BYRON CALHOUN1, GARY DAVIS1, TROY PATIENCE1, MARY BATTAGLIA1, ROBERT E. RICKS1, 1Madigan Army Medical Center, Maternal-Fetal Medicine, Rockford, IL OBJECTIVE: To analyze the impact of participation in a maternal exercise program of moderate intensity on birthweight among pregnant soldiers and matched controls. STUDY DESIGN: Cohort study of birthweight, gestational age, and obstetrical complications among sequential delivered Pregnancy Soldier Wellness Program (PSWP) participants compared with Active Duty (AD) nonparticipants and Family Members (FM). Sub-analysis of primiparous soldiers were matched by ethnicity, smoking and diabetes. ANOVA and Chi-Square used; p < 0.05 as significance. IRB approved study. RESULTS: The Pregnancy Soldier Wellness Program at Fort Lewis has been a sustained program since 1998. During the study period, 1999-2000, 312 PSWP participants delivered and were compared with 206 AD and 522 FM controls selected as next birth of similar age, parity and gestational age. Preterm births, 39 v 25 v 87; were NS. Small for gestation age, 11 v 5 v 9 were NS. BW greater than 4000, 16 v 40 v 98 were p = 0.005. Average birth weight for term black primiparous, 3272 v 3455 v 3566; p = 0.023. Term white primiparous, 3335 v 3533 v 3620; p = 0.001. Shoulder dystocia/fourth degrees were significantly less common among term PSWP; 1/95 v 5/78 v 13/134; p = 0.04. CONCLUSION: Birthweights were 290 grams lighter for participants, but suffered no increase risk of intrauterine growth retardation or prematurity. Participation in the Pregnancy Soldier Wellness Program enhances soldier health with less pelvic floor injury. No adverse perinatal consequences were identified to among participants, rather exercise significantly reduces macrosomia and maternal birth trauma. Corporate wellness programs might consider adding maternal fitness to enhance women’s pelvic floor health.
490
EXPRESSION OF THE GLUCOCORTICOID RECEPTOR IN THE HUMAN PLACENTA THROUGHOUT GESTATION MEN-JEAN LEE1, HERMAN YEE2, SETH GULLER3, NICOLE SWENSON1, REBECCA BAERGEN4, MICHAEL GARABEDIAN5, 1New York University, Obstetrics and Gynecology, New York, NY 2New York University, Pathology, New York, NY 3New York University, Obstetrics and Gynecology; Biochemistry, New York, NY 4Cornell University Medical College, Pathology, New York, NY 5New York University, Microbiology, New York, NY OBJECTIVE: The purpose was to quantify glucocorticoid receptor (GR) expression and activity in the human placenta across gestation using total and phosphorylation site specific antibodies that are surrogates for transcriptional activation and recycling of the receptor. STUDY DESIGN: Paraffin-embedded tissue blocks of placentas from first through third trimesters of pregnancy were retrieved for immunohistochemical analysis. Six first trimester specimens and 10 second-third trimester placentas from appropriately grown pregnancies were examined. The placentas were stained for total human GR, phosphorylated GR at serines 211 (activated), and 226 (recycling). Immunoreactivity was scored based on percentage of positive staining, per cell type, per high powered field. Regression analysis was performed for GR expression by gestational age, with ANOVA, and p < 0.05 considered significant. RESULTS: Total GR expression decreased with advancing gestational age in all placental cell types (p < 0.007). This was associated with significant downregulation of recycling GR in syncytiotrophoblasts (p < 0.03) compared to minor changes in activated GR in the endothelial cells of the maturing placenta. (p < 0.08).
UmA gases, UF, UmA PI and eNOS (densitometry units) Results Control pO2 O2Sat (%) UF (ml/min/kg) UmA eNOS UmV eNOS UtA eNOS UmA PI
488
19.1 49.63 176.4 2862 2099 207 1.18
± 0.7 ± 2.08 ± 13.3 ± 461 ± 235 ± 11.3 ± 0.7
IUGR 10.9 ± 17.87 ± 129.4 ± 4058 ± 3213 ± 241 ± 1.47 ±
1.2 3.49 14.8 288 449 7.08 0.11
P value < 0.001 < 0.001 0.045 0.053 0.053 0.025 0.036
REVERSAL OF REDUCED UTEROPLACENTAL BLOOD FLOW BY SILDENAFIL (VIAGRA) IN AN OVINE MODEL KRISTIN COPPAGE1, ANGELLA FRIEDMAN1, R. SCOTT BAKER1, KENNETH CLARK1, 1University of Cincinnati, Obstetrics and Gynecology, Cincinnati, OH OBJECTIVE: Uteroplacental blood flow (UPBF) increases during pregnancy as a result of nitric oxide production and the resultant vasodilation mediated by cyclic GMP (cGMP). cGMP is broken down by phosphodiesterase 5 (PDE5) which is blocked by Sildenafil, a PDE5 inhibitor. We hypothesized that Sildenafil would: 1) increase UPBF and 2) would reverse the vasoconstriction produced by Serotonin, a selective uterine vasoconstrictor. STUDY DESIGN: Pregnant sheep at 110 days gestation (term = 145) were instrumented. Post recovery sheep whose fetuses had a PaO2 >18 mmHg and pH >7.30 were studied between days 118-138. Three experiments were performed: 1) systemic Sildenafil 2) local uterine artery (UA) Sildenafil and 3) local UA Sildenafil with systemic Serotonin. Changes in maternal/fetal hemodynamics were evaluated. RESULTS: Systemic Viagra (50mg, 100mg) resulted in a 29% decrease in mean arterial pressure (MAP), with minimal changes in maternal heart rate (MHR) and UPBF. The fetus showed an 18% decrease in arterial pressure (FAP), 9% increase in heart rate (FHR) and 20% decrease in umbilical flow (UmbF). Similar fetal results were seen with direct fetal administration of 30ug of Viagra. Local UA administration of Viagra (0.1mg, 0.3 mg, 1mg, 3mg) was associated with small changes in MAP (5,0,6,8%) and MHR (5,13,11,15%) however UPBF did not change. FAP decreased (11,5,13,11%), FHR increased (8,16,-,13%) while UmbF did not change (6,5,6,4%). Serotonin administration reduced UPBF by 28%. Administration of Viagra (0.1mg, 0.3mg, 1mg) increased UPBF by 41, 83 and 75% returning it towards the original baseline. CONCLUSION: In a normal ovine pregnancy, systemic and local Sildenafil does not increase UPBF since the uterine vasculature is maximally dilated, however it does cause maternal/fetal hemodynamic changes. When UPBF is reduced by Serotonin, Sildenafil reverses the vasoconstriction suggesting it may be useful in treating diseases associated with reduced uteroplacental perfusion.
CONCLUSION: GR expression decreases with advancing gestational age in the human placenta. GR downregulation and transcriptional regulation exhibits cell type specificity and may signal a maturational process preceding labor.
Volume 189, Number 6 Am J Obstet Gynecol 487
ENDOTHELIAL NITRIC OXIDE SYNTHASE IN UTEROPLACENTAL VASCULATURE IN AN OVINE MODEL OF IUGR ALESSANDRA PADOAN1, TIMOTHY R.H. REGNAULT2, BARBRA DEVRIJER2, SEAN W. LIMESAND2, RUSSELL V. ANTHONY2, ENRICO FERRAZZI1, RANDALL B. WILKENING2, HENRY L. GALAN3, 1University of Milan, Milan, Italy 2University of Colorado Health Sciences Center, Pediatrics, aurora, CO 3University of Colorado Health Sciences Center, Obstetrics and Gynecology, Denver, CO OBJECTIVE: Endothelial nitric oxide synthase (eNOS) modulates blood flow and is upregulated by hypoxia. Hypothesis: eNOS is upregulated in the uterine artery (UtA), umbilical artery (UmA) and umbilical vein (UmV) in an ovine model of IUGR. STUDY DESIGN: 6 ewes were exposed to hyperthermic conditions from 35115 dGA to induce IUGR and then placed with 6 control ewes in ambient conditions. At 125dGA UmA Doppler pulsatility indices (PI) were assessed and catheters placed for umbilical blood flow (UF) and arterial gas measurements prior to sacrifice at 135dGA. UmA, UmV and UtA were prepared for eNOS Western blot analysis. Data (mean ± SE) were analyzed with t-tests and regression analysis. RESULTS: IUGR birthweights were reduced compared to controls (3.1 ± 1.8kg v 1.5 ± 1.6kg; p < 0.01). See table for results. No differences were seen for pCO2 and pH. UmA PI and UmA [eNOS} correlated well (R = 0.6). CONCLUSION: eNOS protein is largely increased in all vessels, which may be due to a hypoxic uteroplacental environment. Hypoxia, increased placental vascular resistance (UmA PI) and thus shear stress, may account for eNOS upregulation in the UmA and it’s placental branches.(supported by WRHR grant).
489
BIRTHWEIGHT AS A FUNCTION OF MATERNAL DUTY STATUS AND PARTICIPATION IN MANDATORY PHYSICAL TRAINING PROGRAM RODERICK HUME1, BYRON CALHOUN1, GARY DAVIS1, TROY PATIENCE1, MARY BATTAGLIA1, ROBERT E. RICKS1, 1Madigan Army Medical Center, Maternal-Fetal Medicine, Rockford, IL OBJECTIVE: To analyze the impact of participation in a maternal exercise program of moderate intensity on birthweight among pregnant soldiers and matched controls. STUDY DESIGN: Cohort study of birthweight, gestational age, and obstetrical complications among sequential delivered Pregnancy Soldier Wellness Program (PSWP) participants compared with Active Duty (AD) nonparticipants and Family Members (FM). Sub-analysis of primiparous soldiers were matched by ethnicity, smoking and diabetes. ANOVA and Chi-Square used; p < 0.05 as significance. IRB approved study. RESULTS: The Pregnancy Soldier Wellness Program at Fort Lewis has been a sustained program since 1998. During the study period, 1999-2000, 312 PSWP participants delivered and were compared with 206 AD and 522 FM controls selected as next birth of similar age, parity and gestational age. Preterm births, 39 v 25 v 87; were NS. Small for gestation age, 11 v 5 v 9 were NS. BW greater than 4000, 16 v 40 v 98 were p = 0.005. Average birth weight for term black primiparous, 3272 v 3455 v 3566; p = 0.023. Term white primiparous, 3335 v 3533 v 3620; p = 0.001. Shoulder dystocia/fourth degrees were significantly less common among term PSWP; 1/95 v 5/78 v 13/134; p = 0.04. CONCLUSION: Birthweights were 290 grams lighter for participants, but suffered no increase risk of intrauterine growth retardation or prematurity. Participation in the Pregnancy Soldier Wellness Program enhances soldier health with less pelvic floor injury. No adverse perinatal consequences were identified to among participants, rather exercise significantly reduces macrosomia and maternal birth trauma. Corporate wellness programs might consider adding maternal fitness to enhance women’s pelvic floor health.
490
EXPRESSION OF THE GLUCOCORTICOID RECEPTOR IN THE HUMAN PLACENTA THROUGHOUT GESTATION MEN-JEAN LEE1, HERMAN YEE2, SETH GULLER3, NICOLE SWENSON1, REBECCA BAERGEN4, MICHAEL GARABEDIAN5, 1New York University, Obstetrics and Gynecology, New York, NY 2New York University, Pathology, New York, NY 3New York University, Obstetrics and Gynecology; Biochemistry, New York, NY 4Cornell University Medical College, Pathology, New York, NY 5New York University, Microbiology, New York, NY OBJECTIVE: The purpose was to quantify glucocorticoid receptor (GR) expression and activity in the human placenta across gestation using total and phosphorylation site specific antibodies that are surrogates for transcriptional activation and recycling of the receptor. STUDY DESIGN: Paraffin-embedded tissue blocks of placentas from first through third trimesters of pregnancy were retrieved for immunohistochemical analysis. Six first trimester specimens and 10 second-third trimester placentas from appropriately grown pregnancies were examined. The placentas were stained for total human GR, phosphorylated GR at serines 211 (activated), and 226 (recycling). Immunoreactivity was scored based on percentage of positive staining, per cell type, per high powered field. Regression analysis was performed for GR expression by gestational age, with ANOVA, and p < 0.05 considered significant. RESULTS: Total GR expression decreased with advancing gestational age in all placental cell types (p < 0.007). This was associated with significant downregulation of recycling GR in syncytiotrophoblasts (p < 0.03) compared to minor changes in activated GR in the endothelial cells of the maturing placenta. (p < 0.08).
UmA gases, UF, UmA PI and eNOS (densitometry units) Results Control pO2 O2Sat (%) UF (ml/min/kg) UmA eNOS UmV eNOS UtA eNOS UmA PI
488
19.1 49.63 176.4 2862 2099 207 1.18
± 0.7 ± 2.08 ± 13.3 ± 461 ± 235 ± 11.3 ± 0.7
IUGR 10.9 ± 17.87 ± 129.4 ± 4058 ± 3213 ± 241 ± 1.47 ±
1.2 3.49 14.8 288 449 7.08 0.11
P value < 0.001 < 0.001 0.045 0.053 0.053 0.025 0.036
REVERSAL OF REDUCED UTEROPLACENTAL BLOOD FLOW BY SILDENAFIL (VIAGRA) IN AN OVINE MODEL KRISTIN COPPAGE1, ANGELLA FRIEDMAN1, R. SCOTT BAKER1, KENNETH CLARK1, 1University of Cincinnati, Obstetrics and Gynecology, Cincinnati, OH OBJECTIVE: Uteroplacental blood flow (UPBF) increases during pregnancy as a result of nitric oxide production and the resultant vasodilation mediated by cyclic GMP (cGMP). cGMP is broken down by phosphodiesterase 5 (PDE5) which is blocked by Sildenafil, a PDE5 inhibitor. We hypothesized that Sildenafil would: 1) increase UPBF and 2) would reverse the vasoconstriction produced by Serotonin, a selective uterine vasoconstrictor. STUDY DESIGN: Pregnant sheep at 110 days gestation (term = 145) were instrumented. Post recovery sheep whose fetuses had a PaO2 >18 mmHg and pH >7.30 were studied between days 118-138. Three experiments were performed: 1) systemic Sildenafil 2) local uterine artery (UA) Sildenafil and 3) local UA Sildenafil with systemic Serotonin. Changes in maternal/fetal hemodynamics were evaluated. RESULTS: Systemic Viagra (50mg, 100mg) resulted in a 29% decrease in mean arterial pressure (MAP), with minimal changes in maternal heart rate (MHR) and UPBF. The fetus showed an 18% decrease in arterial pressure (FAP), 9% increase in heart rate (FHR) and 20% decrease in umbilical flow (UmbF). Similar fetal results were seen with direct fetal administration of 30ug of Viagra. Local UA administration of Viagra (0.1mg, 0.3 mg, 1mg, 3mg) was associated with small changes in MAP (5,0,6,8%) and MHR (5,13,11,15%) however UPBF did not change. FAP decreased (11,5,13,11%), FHR increased (8,16,-,13%) while UmbF did not change (6,5,6,4%). Serotonin administration reduced UPBF by 28%. Administration of Viagra (0.1mg, 0.3mg, 1mg) increased UPBF by 41, 83 and 75% returning it towards the original baseline. CONCLUSION: In a normal ovine pregnancy, systemic and local Sildenafil does not increase UPBF since the uterine vasculature is maximally dilated, however it does cause maternal/fetal hemodynamic changes. When UPBF is reduced by Serotonin, Sildenafil reverses the vasoconstriction suggesting it may be useful in treating diseases associated with reduced uteroplacental perfusion.
CONCLUSION: GR expression decreases with advancing gestational age in the human placenta. GR downregulation and transcriptional regulation exhibits cell type specificity and may signal a maturational process preceding labor.