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67 The effect of prenatal pravastatin treatment on alteredetal programming of postnatal growth and metabolicunction in a preeclampsia-like murine model

Mollie McDonnold1, Esther Tamayo1, Talar Kechichian1, Phyllisamble1, Monica Longo1, George Saade1, Maged Costantine1

1University of Texas Medical Branch, Department of Obstetrics andynecology, Galveston, TX

OBJECTIVE: Preeclampsia creates an adverse intrauterine environmenthich alters fetal programming and results in long term metabolic

onsequences in the offspring. Pravastatin has been shown to preventreeclampsia in animal models. Our aim was to characterize the ef-

ects of preeclampsia on fetal programming of adult growth and met-bolic function, and evaluate the role of preventive pravastatin ther-py, using a well characterized murine model.

STUDY DESIGN: On day 8 of gestation, CD-1 mice were injectedhrough the tail vein with Adv-sFlt-1 and randomly allocated to prav-statin (5 mg/kg/day; sFlt-prav) or water (sFlt) until weaning. A con-rol group was injected with adenovirus carrying the murine immu-oglobulin G2� Fc fragment (mFc). Offspring (10-14/group) weight,

fasting insulin and cholesterol concentrations were obtained up to 6months of age. IP glucose tolerance testing (IGTT) was performedafter 16 hours of fasting. 2-way ANOVA and ANCOVA were used forstatistical analyses.RESULTS: Pravastatin treatment decreased maternal sFlt-1 concentra-ions in the sFlt group to levels similar to mFc control (p�0.005). Theeight of sFlt offspring was lower than mFc control (p�0.001) until 2onths of age for females and 5 months of age for males. There were

o differences in postnatal growth between mFc and sFlt-pra off-pring. Cholesterol concentrations correlated with offspring weightp�0.02, R2�0.57, ANCOVA), and were similar between groupsTable). At 3 months, sFlt offspring had higher fasting glucosep�.008), but lower insulin (p�0.01) compared with mFc and sFlt-ra offspring. sFlt females had higher peak glucose and higher areander (AUC) the IGTT curve compared with mFc and sFlt-pra at 3nd 6 months.

CONCLUSION: Preeclampsia alters postnatal growth and metabolicfunction in the adult offspring in this animal model. Maternal therapywith pravastatin prevents these alterations in the offspring, providinga potential new opportunity for fetal intervention.

68 Preeclampsia and adverse long term health: aonsequence or a mere unmasking of a predisposition

Egle Bytautiene1, Nataliya Bulayeva2, Geeta Bhat1,i Li2, Kevin Rosenblatt2, George Saade1

1The University of Texas Medical Branch, Obstetrics & Gynecology,alveston, TX, 2The University of Texas Health Sciences Center at Houston,

nstitute of Molecular Medicine, Houston, TXOBJECTIVE: Preeclampsia (PE) is associated with long-term adverse

aternal health such as cardiovascular (CVD) and metabolic diseases.t is also associated with an increase in antiangiogenic factors, such asFlt-1. The objective of this study was to determine whether PE in aell characterized animal model induced by over-expression of sFlt-1

esults in alterations in the maternal circulating proteome that persistong after delivery.

STUDY DESIGN: CD-1 mice at day 8 of gestation were injected withadenovirus carrying sFlt1 or the murine IgG2� Fc as control. Sixmonths after delivery depleted maternal plasma was analyzed by la-

Summary of metabolic findings

bel-free liquid chromatography-mass spectrometry assay. The tan-

S40 American Journal of Obstetrics & Gynecology Supplement to JANUARY 20

dem MS data were searched against mouse database and the resultantintensity data were used to compare abundance of proteins acrossdisease/control plasma pool (MaxQuant/Mascot). Data were ana-lyzed using Ingenuity Pathways Analysis (IPA). Right-tailed Fisher’sexact test was used to calculate a p-value.RESULTS: Out of 150 proteins common for both groups, IPA deter-

ined 97 proteins ready for analysis, out of which 30 were downregu-ated and 67 upregulated. Diseases/disorders analysis showed signifi-ant enrichment of proteins associated with CVD. Out of this clusterhe most abundant proteins were associated with vascular disease20), atherosclerosis (14) and atherosclerotic lesions (12). Other topisease clusters were inflammatory response, organismal injury andbnormalities, hematological and metabolic disease (Fig). Analysisdentified 38 genes as biomarkers for variety of diseases from CVD toancer.

CONCLUSION: Exposure to sFlt1-induced preeclampsia alters multiplebiological functions in mothers that persist later in life. Our resultssuggest that some of the long term adverse outcomes associated withpreeclampsia may actually be a consequence rather than a mere un-masking of an underlying predisposition. If confirmed in humans,developing preventive strategies for preeclampsia should also im-prove long term maternal health.

69 Magnesium sulfate attenuates maternal IL-1�Hima Tam Tam1, Burton Rochelson1, Oonagh Dowling2, Kiran

am Tam3, Xiangying Xue2, Dawnette Lewis4, Christine Metz2

1Hofstra- North Shore School of Medicine, Maternal Fetal Medicine,anhasset, NY, 2Feinstein Institute for Medical Research, Laboratory ofedicinal Biochemistry, Manhasset, NY, 3Baylor College of Medicine,aternal Fetal Medicine, Houston, TX, 4Long Island College Hospital,

Maternal Fetal Medicine, Brooklyn, NYOBJECTIVE: To determine whether the maternal and fetal anti-inflam-

atory effects of Magnesium Sulfate (MgSO4) are mediated viaL-1�.

STUDY DESIGN: On GD 19 Sprague-Dawley rats (E19) were random-zed to 3 groups: saline-saline-saline (SSS); saline-LPS-saline (SLS)nd MgSO4-LPS-MgSO4 (MLM). The dams were given either saliner MgSO4 (270 mg/kg load then 27mg/kg) SQ q20 minutes for 4ours before and 1 ½ hours after i.p injection of either LPS or saline1mg/kg). The dams were euthanized 1 1/2 hours after i.p injection athich time maternal blood (MB), amniotic fluid (AF), fetal blood

FB), fetal brain (BR) and placenta (PLA) were collected. The speci-ens were analyzed for IL-1� using Luminex X-Map technology.

Protein levels for the fetal brain and placental samples were deter-mined using Bio-Rad assay. Data were analyzed using ANOVA withmultiple comparisons. The brain and placenta cytokine levels wereadjusted for protein concentrations.RESULTS: IL-1� was significantly induced in the maternal blood 1 ½hours post LPS administration (31.2 � 6.8 pg/ml (SSS) vs. 419.6 �60.1 pg/ml (SLS); p�0.001). The increase in IL-1� was significantlyattenuated by the administration of MgSO4 (208.9 � 30.1 pg/mlMLM) vs. (SLS) p�0.01). There were no significant differences inL-1� levels within fetal blood, amniotic fluid, placenta and fetal brain

1 ½ hours between the SLS and MLM groups (table 1).CONCLUSION: IL-1� is known to be early mediator of the inflammatorycascade and several studies have shown the induction of this cytokine

Top 5 disease/disorders protein clusters identifiedby IPA according to a score (-log of the p-value)

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www.AJOG.org Hypertension/Physiology Oral Concurrent Session 6

tic pathways by which maternal administered Magnesium Sulfate at-tenuates fetal inflammation. In this study we have demonstratedIL-1� attenuation may be an important pathway by which Magne-ium Sulfate exerts its anti-inflammatory effects in the maternal butot in the fetal compartment. Further studies are needed to betterlucidate this pathway. Research supported by the Oxenhorn family.

70 Altered fetal brain programming in a preeclampsia-likeouse model and its prevention by prenatal

reatment with pravastatinAlissa Carver1, Maged Costantine1, Esther Tamayo1, Huaizhi Yin1,

Regino Perez-Polo2, George Saade1

1University of Texas Medical Branch, Obstetrics & Gynecology, Galveston,X, 2University of Texas Medical Branch, Biochemistry & Molecular Biology,

Galveston, TXOBJECTIVE: Preeclampsia alters fetal programming and results inong-term outcomes in the offspring. Pravastatin, a hydrophilic drughat does not cross the placenta, prevents preeclampsia in animal

odels. This study aims to describe the role of preeclampsia and prav-statin therapy on fetal brain programming using a mouse model.

STUDY DESIGN: At gestation day 8, pregnant CD-1 mice were random-zed to tail vein injection with adenovirus carrying soluble Fms-likeyrosine kinase 1 (sFlt-1) or murine immunoglobulin G2Fc (mFc;ontrol). sFlt-1 dams received pravastatin (5mg/kg/d; sFlt-pra group)r water (sFlt group) until weaning, and mFc received only water

IL-1� levels within the maternaland fetal compartments

Maternal Blood

Fetal Blood

Amniotic Fluid

Fetal Brain Placenta

SSS 31.2 +/- 6.8

126.8 +/- 29.6

100.4 +/- 12.6

0.014 +/- 0.002

0.090 +/- 0.017

SLS 419.6 +/- 60.1

262 +/- 20.7

159.7 +/- 16.6

0.051 +/- 0.014

0.146 +/- 0.030

MLM 208.9 +/- 30.1

241.7 +/- 72.9

158.5 +/- 15.1

0.031 +/- 0.0158

0.168 +/- 0.110

The data are represented as means �/� SEM. The concentrations withinthe maternal blood, fetal blood and amniotic fluid are in pg/ml and theplacenta and fetal brains are in pg/�g of protein.Groups are SSS, saline-saline-saline; SLS, saline-LPS-saline; MLM, MgSO4-LPS-MgSO4.

*Indicates P value � 0.05 SLS vs SSS whereas; **Indicates P value � 0.05 SLS vs MLM.

mFc group). Offspring (n�10-11/group) were sacrificed at

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3-months of age. RNA was extracted from the frontal cortex (FrC),hypothalamus (HT), and cerebellum. Quantitative RT-PCR was usedto measure mRNA expression along pathways involved in apoptosis(Glial fibrillary acidic protein, GFAP), neuronal migration (Reelin),hypoxia (Hypoxia-induced factor 1-alpha), oxidative stress (Cycloox-ygenase 2; Inducible nitric oxide synthase), and myelin and its relatedinflammation (Myelin basic protein (MBP); Amyloid precursor pro-tein). One or two-way ANOVA were used as appropriate for statisticalanalysis.RESULTS: GFAP and reelin expression were up-regulated 2- and 4-foldn the FrC of sFlt offspring compared with control (p�0.025 and.006 respectively). MBP expression was up-regulated 4-fold in sFltffspring HT (p�0.001). Prenatal pravastatin therapy restored ex-ression to levels similar to control (Figure). No gender effect waseen. Expression was similar between groups for all other genes and inhe cerebellum.

CONCLUSION: Preeclampsia alters developmental programming ofpathways for apoptosis, neuronal migration, and myelin inflamma-tion in the frontal cortex and hypothalamus. Maternal therapy withpravastatin restores the normal development of these pathways.

mRNA expression at 3 months of age

ent to JANUARY 2013 American Journal of Obstetrics & Gynecology S41