61 Melatonin supplementation prevents endothelial cell activation, possible relevance to preeclampsia

Abstracts / Pregnancy Hypertension: An International Journal of Women’s Cardiovascular Health 6 (2016) 178–252

207

Table: Magnesium sulfate in preeclampsia/eclampsia in public health system in the municipality of Campinas, SP Component

Facilitator

Legislation

⎫- Healthcare Pact of 2006 rule nº 399/MH ⎫ - Stork Network of 2011 – rule nº 1459/ MH ⎫ - U/E Network - rule nº 1600/ MH ⎫ - RENAME:

REGISTRATION

⎫- Drug registration ANVISA

Barrier - Application is only hospital ⎫- Level for application is missing ⎫ - MgSO4 20% and 50% is included in list drugs for hospital units ⎫ - No prioritizes maternal care ⎫ - No specification in preeclampsia/eclampsia ⎫ - Municipal list: no reference

PROTOCOL

⎫ - Urgency/emergency protocol /2000 - MH ⎫ - High-risk pregnancy protocol /2012 –MH ⎫ - Basic Life Support (BLS) and Advanced Life Support (ALS) / MH

CONDITIONS OF APPLICABILITY

⎫ - All

AVAILABILITY

⎫- UC/ ER and Maternity

⎫ - PHU / EMCS

EQUIPMENT

⎫ - Available at UC, EMCS and Maternity

⎫ - PHU: essential equipment

PHYSICAL

⎫ - Distribution apparently adequate ⎫ - PHU with medication room ⎫ - UC, EMCS and Maternity with adequate architecture

ORGANIZATIONAL

⎫ - Presence of physicians

⎫ - Municipal: lacking ⎫ - Level for application is missing ⎫ - No specification for use in PE/E

⎫ - PHU: to ambulatory scheduling ⎫ - Insufficient number for UC/ER in PHU ⎫ - Exclusion of UC/ER in the care ⎫ - Protocol lacking in UC/ER, ⎫ - Exclusive use by the obstetrician ⎫ - Responsibility not established ⎫ - Elective work ⎫ - Lack of training

⎫ Legend: Rename: National Drug List; ANVISA: National Sanitary Surveillance Agency, EMCS: Emergency Mobile Care Service, UC: urgent care, ER: emergency room, PHU: primary health unit, PE: preeclampsia, E: eclampsia.

doi:10.1016/j.preghy.2016.08.142

Basic science 61 Melatonin supplementation prevents endothelial cell activation, possible relevance to preeclampsia Endothelial dysfunction, anti-angiogenic factors Qi Chen a, Minzhi Zhao b, Mancy Tong a, lance Xu a, Katie Groom a, Peter Stone a, Larry Chamley a (a The University of Auckland, Auckland, New Zealand, b Fudan University, Shanghai, China)

Introduction: Preeclampsia, a human pregnancy-specific disease, is a leading cause of maternal and perinatal mortality and morbidity. It is characterised by maternal endothelial cell dysfunction. Although the exact causes of this disease are unclear, it is thought that there is an increase in oxidative stress in the placenta and leading to the release of a toxic factor (s) from the placenta that triggers maternal endothelial cell activation. Endothelial cell activation precedes the clinical signs and symptoms of the maternal syndrome of preeclampsia. Trophoblast debris, which is shed from the placenta into the maternal blood, is hypothesised to be one such trigger. Our previous studies showed that treating placentae with antiphospholipid antibodies (aPL), which increase the risk of women developing preeclampsia 10 fold [1, 2], or preeclamptic sera, results in the

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production of toxic trophoblastic debris which induces endothelial cell activation [3]. Melatonin is a lipid soluble hormone originally found to be released by the pineal gland but which has been shown to have antioxidant activities and to be produced in the placenta. Given the action of melatonin in regulating oxidative stress and cell death, we investigated whether melatonin supplementation can reverse the effects of preeclamptic sera or aPL on placental oxidative damage and the production of toxic trophoblastic debris. Methods: First trimester placental explants were cultured with aPL (20lg/ml) or preeclamptic sera (n = 6) in the presence or absence of melatonin (1 lM, 10 lM) for 24 h. Trophoblastic debris was collected from the explants and then exposed to monolayers of endothelial cells (HMEC-1) for 24 h and cell surface ICAM-1 expression was measured by cell-based ELISA to determine endothelial cell activation. The expression of nitrotyrosine in placental explants that were treated with aPL or preeclamptic sera was measured by immunohistochemistry or western blotting. Results: 1) Trophoblastic debris from placental explants that were treated with aPL or preeclamptic sera (n = 6) significantly (p < 0.05, respectively) increased the expression of ICAM-1 compared to control-treated endothelial cells. 2) In contrast, trophoblastic debris from explants that were treated with aPL or preeclamptic sera in the presence of melatonin (n = 6) did not significantly increased the expression of ICAM-1 compared to control-treated endothelial cells 3) The expression of nitrotyrosine was significantly increased in placentae that had been treated with aPL or preeclamptic sera. However, this increase in levels of nitrotyrosine was not found in placental explants where aPL or PE sera were added in the presence of melatonin. Conclusions: Our results suggest that there is oxidative damage induced in placentae by either aPL or preeclamptic sera and consequently trophoblastic debris produced by these placentae is more toxic to endothelial cells. Melatonin can reverse this placental oxidative damage and reduced the toxicity of the trophoblastic debris. Since the pathogenesis of preeclampsia is thought to be partially dependent on the maternal response to placental factors such as toxic trophoblastic debris shed from placentae, our results encouraging suggest that melatonin might be a useful tool to alleviate the clinical manifestations of preeclampsia. doi:10.1016/j.preghy.2016.08.143

Clinical science 62 Angiotensin II receptor type 1 autoantibodies (AT1AA) in preeclampsia: Results of a retrospective longitudinal analysis Angiogenic factors Shikha Aggarwal, Angela Makris, Annemarie Hennessy (School of Meidicine, Western Sydney University, Sydney Australia) Introduction: AT1AA have been shown to play a role in endothelial dysfunction, which is a key factor in the pathogenesis of PE. AT1-AA have been shown to mediate disease and thus may have a role as a biomarker. Objectives: Assess the longitudinal changes and utility of AT1-AA as a biomarker in hypertensive disorders of pregnancy (gestational hypertension [GHT] and preeclampsia [PE]). Methods: An observational, longitudinal case control study of an unselected population was undertaken. Plasma samples were collected at weeks 12, 18, 28, 36, 40 and 6 weeks post-partum. Baseline clinical, demographic data and pregnancy outcomes were

prospectively collected. Pregnancy outcomes included normotensive (control), GHT or PE. AT1-AA were measured using a validated commercial ELISA (One-Lambda). Statistical analyses were carried out using SPSS, data expressed as median (interquartile range] and statistical significance was set at p <0.05. Result: 351 women were recruited, 17 and 18 developed GHT and PE respectively. AT1AA levels were measured in 76 women. Maternal age was similar for all three groups. There was a statistically significant difference in maternal weight (p = 0.015), height (p = 0.046) and BMI (p = 0.041) between groups (Kruskal-Wallis test). Baseline systolic and diastolic blood pressure was significantly different across groups with p values of 0.029 and 0.006 respectively (Kruskal–Wallis). The PE group had higher rate of preeclampsia history 28.1% compared to controls 1.3% p <0.01. There was no significant difference in the rate of smoking, primiparity and gestational diabetes mellitus (GDM). AT1-AA levels decreased from 12 weeks 15.44[12.77–17.86] u/ml to term 11.53[5.75–17.78] u/ml but this was not statistically significant. There was no difference in AT1AA between the three diagnostic groups at any time point (Kruskal– Wallis analysis analysis p >0.05) (Fig. 1). AT1AA at 12 weeks had a positive correlation with serum PAPP-A (p = 0.008) and BHCG (p = 0.04) but no relationship with other angiogenic markers at 12 weeks such as soluble fms like tyrosine kinase receptor 1, plasma like growth factor or serum endogolin. AT1AA at 12 weeks was not associated with later preeclampsia (p = 0.25). Conclusions: AT1AA are not specific for PE and do not prove to be biomarkers of disease. doi:10.1016/j.preghy.2016.08.144

Basic science 63 Transcriptome wide profiling of mRNA 30 -ends in preeclampsia using PAS-SEQ Genomics Ami Ashar a, Yasin Kaymaz b, Augustine Rajakumar c, Jeffrey A. Bailey b, S. Ananth Karumanchi d, Melissa J. Moore a (a Umass Medical School, Worcester, United States, b Program in Bioinformatics and Integrative Biology and Department of Pathology, University of Massach, MA United States, c Department of Gynecology and Obstetrics, Emory University, GA United States, d Division of Nephrology, Department of Medicine and Center for Vascular Biology Research, MA United States) Maternal symptoms of preeclampsia (PE) are a primary consequence of excessive levels of sFlt1 protein in the maternal blood supply 1. sFlt1 is encoded by the gene Fms-related tyrosine kinase 1 (FLT1), which produces multiple protein isoforms: (1) transmembrane bound Flt1 (mFlt1), a tyrosine kinase receptor participating in vascular endothelial growth factor (VEGF) signaling; and (2) truncated and blood circulating (soluble) forms, sFlt1s, which lack the transmembrane anchoring domain and act as antagonists to mFlt1. Relative production of these isoforms results from polyadenylation sites within exon 30 (mFlt1) or within introns 13 and 14 (sFlt1s). The goals of this project are three-fold: (1) Determine whether increased sFlt1 production in PE results from an overall increase in FLT1 gene expression, from preferential use of intronic polyadenylation sites or both; (2) assess general gene expression changes in PE versus non-PE placentae; (3) determine if there is a global change in APA (alternative polyadenylation) site usage in PE. To address these goals we used a next-generation sequencing specifically targeting the 30 -ends of mRNAs: PolyAdenylation Site