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540: Increased placental apoptosis and increased X-linked inhibitor of apoptosis protein in a baboon model of maternal nutrient restriction
SMFM Abstracts 537
HYPERTENSION IN RESPONSE TO AGONISTIC AUTOANTIBODIES TO THE ANGIOTENSIN II TYPE I RECEPTOR IS ASSOCIATED WITH ELEVATED CIRCULATING SFLT-1 MARC PARRISH1, SYDNEY MURPHY1, RALF DECHEND2, SHARON KEISER1, EDWARD VEILLON1, KATHY COCKRELL1, LILLIAN FOURNIER1, JAMES MARTIN1, BABBETTE LAMARCA1, 1University of Mississippi Medical Center, Jackson, Mississippi, 2HELIOS Clinic, Charite, Campus-Buch & Max-Delbrueck Center, Berlin, Germany OBJECTIVE: Elevated antiangiogenic factors such as soluble fms like tyrosine kinase-1 (sFlt-1), and production of agonistic autoantibody to the angiotensin II type I receptor (AT1-AA) are associated with abnormal placentation as well as hypertension during pregnancy. Previous investigators have suggested that immunoglobulin from preeclamptic women increases sFlt-1 production. The objective of this study was to determine if hypertension in response to purified AT1-AA in pregnant rats increases sFlt-1 production. STUDY DESIGN: Specific AT1-AA was isolated from a transgenic rat strain overexpressing components of the renin angiotensin system. Sprague-Dawley rats on Day 12 of pregnancy were infused with rat AT1-AA (1:50) for 7 days. On day 18, carotid artery catheters were inserted. On day 19 mean arterial pressures(MAP), as well maternal and pup weights were measured. Blood, kidneys and placentas were then collected. Plasma was analyzed for circulating sFlt-1. RESULTS: Infusion of the AT1-AA for 7 days increased mean arterial pressure in normal pregnant (NP) rats from 99⫹/⫺ 1 to 118 ⫹/⫺ 1mmHg (P⬍0.0001). This hypertension was associated with decreased maternal weight gain, (343 ⫹/⫺4 to 322 ⫹/⫺ 5 grams (P⬍0.003)) and heart weight (0.96 ⫹/⫺ 0.01 to 0.92 ⫹/⫺ 0.01 grams(P⬍ 0.04)). Importantly circulating sFlt-1 increased from 245 ⫹/⫺ 38 in NP rats to 3,920 ⫹/⫺ 798 pg/ml in AT1-AA-induced hypertensive rats. CONCLUSION: These data demonstrate that hypertension in response to AT1-AA is associated with increased production of sFlt-1during pregnancy and could be an additional mechanism whereby AT1-AA increases blood pressure.
www.AJOG.org 539
0002-9378/$ - see front matter doi:10.1016/j.ajog.2008.09.567
HEPATIC AND PERIPHERAL INSULIN RESISTANCE OF PREGNANCY ARE NOT REVERSED IN EARLY POSTPARTUM RATS HYE HEO1, LINGGUANG CUI1, RADHIKA H. MUZUMDAR1, DEREK M. HUFFMAN1, NIR BARZILAI1, FRANCINE EINSTEIN1, 1Albert Einstein College of Medicine, Bronx, New York OBJECTIVE: Insulin resistance is a metabolic hallmark of pregnancy and is thought to be quickly reversed with removal of the placenta and placental factors. We previously showed that visceral fat contributes to insulin resistance in pregnancy. We sought to determine if insulin sensitivity was returned in the early postpartum period in the absence of circulating placental factors. STUDY DESIGN: Two groups of age-matched SD rats were studied: Postpartum day #7-8 non-lactating females (PP; n⫽6) and age-matched virgin females (Vir; n⫽6). Using hyperinsulinemic-euglycemic clamp technique, chronically catheterized, awake, unrestrained rats were given infusions of insulin (3 mU䡠kg-1䡠min-1), somatostatin(1.5 ug䡠kg-1䡠min-1) and 3H3-glucose (to follow glucose fluxes). Glucose was clamped at basal levels using 25% dextrose solution. After clamps, adipose samples were collected and weighed. RESULTS: PP weighed significantly more than Vir (266.2⫾4 v 235.7⫾9g, p⬍0.05) and had greater perinephric (1.7⫾0.1 vs.0.8⫾0.1g, p⬍0.01), paraovarian (1.3⫾0.1 vs.0.7⫾0.1g, p⬍0.01) and total visceral fat depots (3.8⫾0.1 vs.2.1⫾0.3g, p⬍0.01). The weights of mesenteric fat depots were similar. The glucose infusion rate (GIR) was lower in PP compared to Vir (7.4⫾0.2 vs.15.7⫾0.2 mg/kg/min, p⬍0.001). Under hyperinsulinemic conditions, hepatic glucose production was significantly greater in PP compared to Vir (10.7⫾0.2 vs.7.4⫾0.4 mg/kg/min (44 v 62% suppression from basal) respectively, p⬍0.001) and peripheral glucose uptake during the clamp (Rd) was lower in PP (18.0⫾0.2 mg/kg/min) compared to Vir (22.4⫾0.3mg/kg/min, p⬍0.01) demonstrating persistent hepatic and peripheral insulin resistance in post partum animals. CONCLUSION: Insulin sensitivity is not restored in the early postpartum period in non-lactating rodents despite the absence of circulating placental factors and underscoring the contribution of pregnancy-related body composition changes. Further, postpartum weight retention may confer increased maternal risk for developing the metabolic syndrome and diabetes later in life. 0002-9378/$ - see front matter doi:10.1016/j.ajog.2008.09.569
538
VIP, ADNP AND GFAP EXPRESSION IN ADULT OFFSPRING BRAIN IN A MODEL OF FETAL ALCOHOL SYNDROME MADDALENA INCERTI1, ROBIN ROBERSON1, JOY VINK2, DANIEL ABEBE1, CATHERINE SPONG1, 1National Institutes of Health (NIH), NICHD, Unit on Perinatal and Developmental Neurobiology, Bethesda, Maryland, 2 Georgetown University, Washington, District of Columbia OBJECTIVE: Fetal alcohol syndrome (FAS) is characterized by neurodevelopmental anomalies with learning impairment and alterations in neuropeptides including decreased VIP immunoreactivity. Prenatal treatment with the peptides NAP⫹SAL prevents the alcohol-induced learning deficits in FAS. VIP regulates ADNP, the parent protein of NAP and acts through glial cells. GFAP is a marker for glial cells. Our objective was to evaluate if adult treatment with NAP⫹SAL affects VIP, ADNP and GFAP expression in adult brain after prenatal alcohol exposure. STUDY DESIGN: C57Bl6/J mice were treated with alcohol (0.03 mL/g) or placebo on gestational day 8 (Webster FAS model). On day 40, in utero alcoholexposed males were treated daily for 10d with NAP⫹SAL(n⫽6) or placebo(n⫽4); and control offspring with placebo(n⫽12) with the treatment blinded. Learning evaluation began after 3d using the Morris watermaze and T-maze where a higher rate of alternation is desirable. The cortex (C) and hippocampus (H) were isolated and calibrator-normalized relative rt-PCR used primers for VIP, ADNP and GFAP with GAPDH standardization. Statistics: ANOVA, Fisher PLSD. RESULTS: Adult treatment with NAP⫹SAL prevented the alcohol-induced learning deficit in the watermaze and T-maze and the ADNP up-regulation in the C, but did not prevent the VIP down-regulation in the H (all P⬍.05). There was no change in expression of ADNP and GFAP in H and VIP in C after alcohol exposure.
540
INCREASED PLACENTAL APOPTOSIS AND INCREASED X-LINKED INHIBITOR OF APOPTOSIS PROTEIN IN A BABOON MODEL OF MATERNAL NUTRIENT RESTRICTION JUAN A ARROYO1, CUN LI2, NATALIA SCHLABRITZ-LOUTSEVITCH2, TOM MCDONALD3, BRADLEY T. ZIEBELL1, PETER NATHANIELSZ2, HENRY GALAN1, 1University of Colorado and Health Sciences Center, Obstetrics and Gynecology, Aurora, Colorado, 2University of Texas Health Science Center at San Antonio, Obstetrics and Gynecology, San Antonio, Texas, 3Southwest Foundation for Biomedical Research, San Antonio, Texas OBJECTIVE: Apoptosis is a component of normal development and differentiation in most tissues. Increased placental apoptosis is normally seen at term in humans and is also associated with complicated pregnancies such as intrauterine growth restriction. Our objective was to determine apoptosis rate in the term placenta of a baboon model of maternal nutrient restriction (MNR). STUDY DESIGN: From 30 to 165 days of gestation (dG) female baboons were fed ad lib (CTR; n⫽7) or 70% CTR diet (MNR; n⫽6) with necropsy and tissue collection at 165 dG. Placental tissue was collected and snap frozen for protein studies. Placental villous apoptosis was determined by the TUNEL assay and confirmed by cytokeratin 18 cleavage via immunohstochemistry. Western blot analysis (50g of total protein) was performed to compare XIAP protein between groups. Actin westerns on the same blots were used to account for inter-lane loading variation. Data are presented as mean ⫾ SE compared with the Wilcoxon rank sum test. RESULTS: Compared to controls, MNR pregnancies demonstrated: 1) reduced placental weight (177.3 ⫾ 7.89g vs. 145.0 ⫾ 7.23 g; p⬍0.05) with no differences in fetal weight, 2) a 2-fold increase (p⬍0.001) in placental villous apoptosis (TUNEL), 3) an increase in villous cytokeratin 18 cleavage, and 4) a 1.5-fold increase (p⬍0.007) in XIAP protein concentration. CONCLUSION: We conclude that placental apoptosis is increased in this baboon model of MNR at term and that the increase in XIAP may be a protective mechanism against this apoptosis. Future studies are needed to delineate the mechanism of apoptosis and to determine if apoptosis occurs earlier in pregnancy in this model of MNR. (Supported by HD21350am and R01 HDO34837-08).
CONCLUSION: Adult treatment with NAP⫹SAL prevented the alcohol-induced learning deficit and the changes in ADNP expression in the C, but not the downregulation of VIP, probably because the peptides are downstream of VIP. 0002-9378/$ - see front matter doi:10.1016/j.ajog.2008.09.568
0002-9378/$ - see front matter doi:10.1016/j.ajog.2008.09.570
S158
American Journal of Obstetrics & Gynecology Supplement to DECEMBER 2008
HYPERTENSION IN RESPONSE TO AGONISTIC AUTOANTIBODIES TO THE ANGIOTENSIN II TYPE I RECEPTOR IS ASSOCIATED WITH ELEVATED CIRCULATING SFLT-1 MARC PARRISH1, SYDNEY MURPHY1, RALF DECHEND2, SHARON KEISER1, EDWARD VEILLON1, KATHY COCKRELL1, LILLIAN FOURNIER1, JAMES MARTIN1, BABBETTE LAMARCA1, 1University of Mississippi Medical Center, Jackson, Mississippi, 2HELIOS Clinic, Charite, Campus-Buch & Max-Delbrueck Center, Berlin, Germany OBJECTIVE: Elevated antiangiogenic factors such as soluble fms like tyrosine kinase-1 (sFlt-1), and production of agonistic autoantibody to the angiotensin II type I receptor (AT1-AA) are associated with abnormal placentation as well as hypertension during pregnancy. Previous investigators have suggested that immunoglobulin from preeclamptic women increases sFlt-1 production. The objective of this study was to determine if hypertension in response to purified AT1-AA in pregnant rats increases sFlt-1 production. STUDY DESIGN: Specific AT1-AA was isolated from a transgenic rat strain overexpressing components of the renin angiotensin system. Sprague-Dawley rats on Day 12 of pregnancy were infused with rat AT1-AA (1:50) for 7 days. On day 18, carotid artery catheters were inserted. On day 19 mean arterial pressures(MAP), as well maternal and pup weights were measured. Blood, kidneys and placentas were then collected. Plasma was analyzed for circulating sFlt-1. RESULTS: Infusion of the AT1-AA for 7 days increased mean arterial pressure in normal pregnant (NP) rats from 99⫹/⫺ 1 to 118 ⫹/⫺ 1mmHg (P⬍0.0001). This hypertension was associated with decreased maternal weight gain, (343 ⫹/⫺4 to 322 ⫹/⫺ 5 grams (P⬍0.003)) and heart weight (0.96 ⫹/⫺ 0.01 to 0.92 ⫹/⫺ 0.01 grams(P⬍ 0.04)). Importantly circulating sFlt-1 increased from 245 ⫹/⫺ 38 in NP rats to 3,920 ⫹/⫺ 798 pg/ml in AT1-AA-induced hypertensive rats. CONCLUSION: These data demonstrate that hypertension in response to AT1-AA is associated with increased production of sFlt-1during pregnancy and could be an additional mechanism whereby AT1-AA increases blood pressure.
www.AJOG.org 539
0002-9378/$ - see front matter doi:10.1016/j.ajog.2008.09.567
HEPATIC AND PERIPHERAL INSULIN RESISTANCE OF PREGNANCY ARE NOT REVERSED IN EARLY POSTPARTUM RATS HYE HEO1, LINGGUANG CUI1, RADHIKA H. MUZUMDAR1, DEREK M. HUFFMAN1, NIR BARZILAI1, FRANCINE EINSTEIN1, 1Albert Einstein College of Medicine, Bronx, New York OBJECTIVE: Insulin resistance is a metabolic hallmark of pregnancy and is thought to be quickly reversed with removal of the placenta and placental factors. We previously showed that visceral fat contributes to insulin resistance in pregnancy. We sought to determine if insulin sensitivity was returned in the early postpartum period in the absence of circulating placental factors. STUDY DESIGN: Two groups of age-matched SD rats were studied: Postpartum day #7-8 non-lactating females (PP; n⫽6) and age-matched virgin females (Vir; n⫽6). Using hyperinsulinemic-euglycemic clamp technique, chronically catheterized, awake, unrestrained rats were given infusions of insulin (3 mU䡠kg-1䡠min-1), somatostatin(1.5 ug䡠kg-1䡠min-1) and 3H3-glucose (to follow glucose fluxes). Glucose was clamped at basal levels using 25% dextrose solution. After clamps, adipose samples were collected and weighed. RESULTS: PP weighed significantly more than Vir (266.2⫾4 v 235.7⫾9g, p⬍0.05) and had greater perinephric (1.7⫾0.1 vs.0.8⫾0.1g, p⬍0.01), paraovarian (1.3⫾0.1 vs.0.7⫾0.1g, p⬍0.01) and total visceral fat depots (3.8⫾0.1 vs.2.1⫾0.3g, p⬍0.01). The weights of mesenteric fat depots were similar. The glucose infusion rate (GIR) was lower in PP compared to Vir (7.4⫾0.2 vs.15.7⫾0.2 mg/kg/min, p⬍0.001). Under hyperinsulinemic conditions, hepatic glucose production was significantly greater in PP compared to Vir (10.7⫾0.2 vs.7.4⫾0.4 mg/kg/min (44 v 62% suppression from basal) respectively, p⬍0.001) and peripheral glucose uptake during the clamp (Rd) was lower in PP (18.0⫾0.2 mg/kg/min) compared to Vir (22.4⫾0.3mg/kg/min, p⬍0.01) demonstrating persistent hepatic and peripheral insulin resistance in post partum animals. CONCLUSION: Insulin sensitivity is not restored in the early postpartum period in non-lactating rodents despite the absence of circulating placental factors and underscoring the contribution of pregnancy-related body composition changes. Further, postpartum weight retention may confer increased maternal risk for developing the metabolic syndrome and diabetes later in life. 0002-9378/$ - see front matter doi:10.1016/j.ajog.2008.09.569
538
VIP, ADNP AND GFAP EXPRESSION IN ADULT OFFSPRING BRAIN IN A MODEL OF FETAL ALCOHOL SYNDROME MADDALENA INCERTI1, ROBIN ROBERSON1, JOY VINK2, DANIEL ABEBE1, CATHERINE SPONG1, 1National Institutes of Health (NIH), NICHD, Unit on Perinatal and Developmental Neurobiology, Bethesda, Maryland, 2 Georgetown University, Washington, District of Columbia OBJECTIVE: Fetal alcohol syndrome (FAS) is characterized by neurodevelopmental anomalies with learning impairment and alterations in neuropeptides including decreased VIP immunoreactivity. Prenatal treatment with the peptides NAP⫹SAL prevents the alcohol-induced learning deficits in FAS. VIP regulates ADNP, the parent protein of NAP and acts through glial cells. GFAP is a marker for glial cells. Our objective was to evaluate if adult treatment with NAP⫹SAL affects VIP, ADNP and GFAP expression in adult brain after prenatal alcohol exposure. STUDY DESIGN: C57Bl6/J mice were treated with alcohol (0.03 mL/g) or placebo on gestational day 8 (Webster FAS model). On day 40, in utero alcoholexposed males were treated daily for 10d with NAP⫹SAL(n⫽6) or placebo(n⫽4); and control offspring with placebo(n⫽12) with the treatment blinded. Learning evaluation began after 3d using the Morris watermaze and T-maze where a higher rate of alternation is desirable. The cortex (C) and hippocampus (H) were isolated and calibrator-normalized relative rt-PCR used primers for VIP, ADNP and GFAP with GAPDH standardization. Statistics: ANOVA, Fisher PLSD. RESULTS: Adult treatment with NAP⫹SAL prevented the alcohol-induced learning deficit in the watermaze and T-maze and the ADNP up-regulation in the C, but did not prevent the VIP down-regulation in the H (all P⬍.05). There was no change in expression of ADNP and GFAP in H and VIP in C after alcohol exposure.
540
INCREASED PLACENTAL APOPTOSIS AND INCREASED X-LINKED INHIBITOR OF APOPTOSIS PROTEIN IN A BABOON MODEL OF MATERNAL NUTRIENT RESTRICTION JUAN A ARROYO1, CUN LI2, NATALIA SCHLABRITZ-LOUTSEVITCH2, TOM MCDONALD3, BRADLEY T. ZIEBELL1, PETER NATHANIELSZ2, HENRY GALAN1, 1University of Colorado and Health Sciences Center, Obstetrics and Gynecology, Aurora, Colorado, 2University of Texas Health Science Center at San Antonio, Obstetrics and Gynecology, San Antonio, Texas, 3Southwest Foundation for Biomedical Research, San Antonio, Texas OBJECTIVE: Apoptosis is a component of normal development and differentiation in most tissues. Increased placental apoptosis is normally seen at term in humans and is also associated with complicated pregnancies such as intrauterine growth restriction. Our objective was to determine apoptosis rate in the term placenta of a baboon model of maternal nutrient restriction (MNR). STUDY DESIGN: From 30 to 165 days of gestation (dG) female baboons were fed ad lib (CTR; n⫽7) or 70% CTR diet (MNR; n⫽6) with necropsy and tissue collection at 165 dG. Placental tissue was collected and snap frozen for protein studies. Placental villous apoptosis was determined by the TUNEL assay and confirmed by cytokeratin 18 cleavage via immunohstochemistry. Western blot analysis (50g of total protein) was performed to compare XIAP protein between groups. Actin westerns on the same blots were used to account for inter-lane loading variation. Data are presented as mean ⫾ SE compared with the Wilcoxon rank sum test. RESULTS: Compared to controls, MNR pregnancies demonstrated: 1) reduced placental weight (177.3 ⫾ 7.89g vs. 145.0 ⫾ 7.23 g; p⬍0.05) with no differences in fetal weight, 2) a 2-fold increase (p⬍0.001) in placental villous apoptosis (TUNEL), 3) an increase in villous cytokeratin 18 cleavage, and 4) a 1.5-fold increase (p⬍0.007) in XIAP protein concentration. CONCLUSION: We conclude that placental apoptosis is increased in this baboon model of MNR at term and that the increase in XIAP may be a protective mechanism against this apoptosis. Future studies are needed to delineate the mechanism of apoptosis and to determine if apoptosis occurs earlier in pregnancy in this model of MNR. (Supported by HD21350am and R01 HDO34837-08).
CONCLUSION: Adult treatment with NAP⫹SAL prevented the alcohol-induced learning deficit and the changes in ADNP expression in the C, but not the downregulation of VIP, probably because the peptides are downstream of VIP. 0002-9378/$ - see front matter doi:10.1016/j.ajog.2008.09.568
0002-9378/$ - see front matter doi:10.1016/j.ajog.2008.09.570
S158
American Journal of Obstetrics & Gynecology Supplement to DECEMBER 2008