PP017. Withdrawn
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PP015. Plasma levels and placental expression of BNP/NT- proBNP in early and late onset preeclampsia Junus Katja, Wikström Anna-Karin, Larsson Anders, Matts Olovsson Introduction: Women with preeclampsia (PE) have ele- vated plasma levels of NT-proBNP. We hypothesized that the placenta may be a source to these elevated levels. Objectives: Our objectives were to study the plasma levels of NT-proBNP and the protein and mRNA expression of placental BNP in women with early and late onset PE and controls. Methods: Plasma levels of NT-proBNP were measured in women with early (n = 18) and late (n = 20) onset PE, in two groups of healthy pregnant women in gestational week 24–32 (n = 22) and 36–42 (n = 14), and in non-pregnant women (n = 20). Placental BNP protein and mRNA was stud- ied with immunohistochemistry and qPCR. Placental release of NT-proBNP was studied with tissue culturing. Results: Women with early (365 (14-9815) pg/ml) and late (176 (33-2547) pg/ml) onset PE had higher levels of NT-proBNP in plasma than their respective controls (p < 0.001). A tendency towards higher plasma levels in early compared to late onset PE was observed (p = 0.057). 20 out of 25 placental tissue samples had proBNP mRNA, no differ- ences between the study groups were found. BNP protein was found in maternal spiral arteries and syncytiotropho- blasts. NT-proBNP peptide (6–7 pg/ml) was present in med- ium used for placenta cultures. Conclusions: Our results suggest that there may be a pla- cental source of NT-proBNP. If this source is responsible for the elevated plasma levels of NT-proBNP in preeclamptic women and what role, if any, BNP/NT-proBNP play in PE pathophysiology remains to be elucidated. doi:10.1016/j.preghy.2013.04.043 PP016. Relation of apoptosis, proliferation and angio- genesis in early and late onset of preeclampsia Khodzhaeva Zulfiya, Kogan Eugenia, Kholin Alexey, Albina Akatyeva, Vavina Olga, Sukhikh Gennady Introduction: The placental bed plays a key role in placen- tation during gestation. Most studies investigated the expression of angiogenic factors in the placenta, but their expression and potential role in the placental bed have not been investigated adequately. Objectives: The aim of the study was to examine the expression of the fact is that Apo-Cas is apoptotic marker and VEGF in placental bed of pregnancy with early, late- onset PE and without PE. Methods: Placental bed biopsy tissues obtained during Cesarean Section from patients with early-onset (n = 15), late-onset (n = 15) and without (n = 15) PE. The normoten- sive controls without PE were matched for gestational age at delivery with patients with PE. The expression of Apo- Cas and VEGF in placental bed tissues were evaluated using reverse transcriptase PCR, real-time PCR, immunohisto- chemistry and Western blot. Results: There was no statistical difference between the PE group and normotensive control group in age and body mass index. The level of apoptotic marker Apo-Cas was higher in early-compared to late-onset of PE (5% ± 1.4, and 15% ± 2.7). The expression of VEGF was significantly decreased in both PE groups compare to control (p < 0.05), but not statistically significant between groups with PE. We also revealed reduc- tion of VEGF receptors in endometrial stroma and its absence in endothelial cells. Conclusion: This study showed the prevalence of apop- tosis and decreased expression of VEGF in the placental bed of pregnancies complicated by PE compared with control. Further research will help to create the pathoge- netic basis for early prediction, recognition and manage- ment of PE. doi:10.1016/j.preghy.2013.04.044 PP018. Does altitude affect the placental renin angio- tensin system (RAS) in pre-eclampsia (PE)? Kurlak Lesia, Mistry Hiten, Cindrova-Davies Tereza, van Patot Martha Tissot, Burton Graham, Pipkin Fiona Broughton Introduction: We have previously shown the activation of placental RAS in high altitude normotensive (NT) pregnan- cies [1], presumably related to hypoxia during placental development. PE incidence is increased at high altitude; hypoxia-reoxygenation injury contributes to the elevated oxidative stress seen in this condition. Binding of Angioten- sin II to its type I receptor (AT1R) increases generation of reactive oxygen species and we have shown an increase in AT1R in PE [2]. Therefore we investigated whether the RAS is further augmented in PE at high altitudes. Methods: Biopsies were collected, with informed, written consent, from NT and PE women (ISSHP guidelines) at sea- level (UK) and high altitude, 3100 m (Leadville, Colorado) immediately after vaginal delivery or elective non-laboured Caesarean section. mRNA expression of the RAS compo- nents (angiotensinogen (AGT), prorenin, prorenin receptor, AT1R and angiotensin type 2 receptor) was determined by qRT-PCR and normalised to 3 stably-expressed house- keeping genes using geNorm. Protein expression was semi-quantitatively assessed by immunohistochemistry (H score) and compared between groups (Mann Whitney U test). Results: mRNA expression of all RAS components was unaf- fected by altitude in NT, but significantly lower for all in PE (P = 0.026 < 0.0001). However, in NT, protein expression of all components except AGT was significantly increased at altitude (P = 0.037 < 0.001). Protein expression of AGT and AT1R was also increased in altitude PE (P < 0.004 for both). Conclusion: We have shown the placental RAS to be acti- vated at altitude, with apparent differences in transcription/ translation. Hypoxia-inducible transcription factors such as HIF-1 enhance transcription and translation of many genes encoding ‘‘rescue’’ proteins; this might explain the effects of altitude in NTs. HIF-1 is increased in PE, which might allow increased protein expression at altitude in spite of lower mRNA. Poster Presentations / Pregnancy Hypertension: An International Journal of Women’s Cardiovascular Health 3 (2013) 67–99 73
Transcript of PP017. Withdrawn
PP015. Plasma levels and placental expression of BNP/NT-proBNP in early and late onset preeclampsiaJunus Katja, Wikström Anna-Karin, Larsson Anders, MattsOlovsson
Introduction: Women with preeclampsia (PE) have ele-vated plasma levels of NT-proBNP. We hypothesized thatthe placenta may be a source to these elevated levels.
Objectives: Our objectives were to study the plasma levels ofNT-proBNP and the protein and mRNA expression of placentalBNP in women with early and late onset PE and controls.
Methods: Plasma levels of NT-proBNP were measured inwomen with early (n = 18) and late (n = 20) onset PE, intwo groups of healthy pregnant women in gestational week24–32 (n = 22) and 36–42 (n = 14), and in non-pregnantwomen (n = 20). Placental BNP protein and mRNA was stud-ied with immunohistochemistry and qPCR. Placental releaseof NT-proBNP was studied with tissue culturing.
Results: Women with early (365 (14-9815) pg/ml) andlate (176 (33-2547) pg/ml) onset PE had higher levels ofNT-proBNP in plasma than their respective controls(p < 0.001). A tendency towards higher plasma levels in earlycompared to late onset PE was observed (p = 0.057). 20 outof 25 placental tissue samples had proBNP mRNA, no differ-ences between the study groups were found. BNP proteinwas found in maternal spiral arteries and syncytiotropho-blasts. NT-proBNP peptide (6–7 pg/ml) was present in med-ium used for placenta cultures.
Conclusions: Our results suggest that there may be a pla-cental source of NT-proBNP. If this source is responsiblefor the elevated plasma levels of NT-proBNP in preeclampticwomen and what role, if any, BNP/NT-proBNP play in PEpathophysiology remains to be elucidated.
doi:10.1016/j.preghy.2013.04.043
PP016. Relation of apoptosis, proliferation and angio-genesis in early and late onset of preeclampsiaKhodzhaeva Zulfiya, Kogan Eugenia, Kholin Alexey,Albina Akatyeva, Vavina Olga, Sukhikh Gennady
Introduction: The placental bed plays a key role in placen-tation during gestation. Most studies investigated theexpression of angiogenic factors in the placenta, but theirexpression and potential role in the placental bed have notbeen investigated adequately.
Objectives: The aim of the study was to examine theexpression of the fact is that Apo-Cas is apoptotic markerand VEGF in placental bed of pregnancy with early, late-onset PE and without PE.
Methods: Placental bed biopsy tissues obtained duringCesarean Section from patients with early-onset (n = 15),late-onset (n = 15) and without (n = 15) PE. The normoten-sive controls without PE were matched for gestational ageat delivery with patients with PE. The expression of Apo-Cas and VEGF in placental bed tissues were evaluated usingreverse transcriptase PCR, real-time PCR, immunohisto-chemistry and Western blot.
Results: There was no statistical difference between the PEgroup and normotensive control group in age and body mass
index. The level of apoptotic marker Apo-Cas was higher inearly-compared to late-onset of PE (5% ± 1.4, and 15% ± 2.7).The expression of VEGF was significantly decreased in bothPE groups compare to control (p < 0.05), but not statisticallysignificant between groups with PE. We also revealed reduc-tion of VEGF receptors in endometrial stroma and itsabsence in endothelial cells.
Conclusion: This study showed the prevalence of apop-tosis and decreased expression of VEGF in the placentalbed of pregnancies complicated by PE compared withcontrol. Further research will help to create the pathoge-netic basis for early prediction, recognition and manage-ment of PE.
doi:10.1016/j.preghy.2013.04.044
PP018. Does altitude affect the placental renin angio-tensin system (RAS) in pre-eclampsia (PE)?Kurlak Lesia, Mistry Hiten, Cindrova-Davies Tereza, vanPatot Martha Tissot, Burton Graham, Pipkin FionaBroughton
Introduction: We have previously shown the activation ofplacental RAS in high altitude normotensive (NT) pregnan-cies [1], presumably related to hypoxia during placentaldevelopment. PE incidence is increased at high altitude;hypoxia-reoxygenation injury contributes to the elevatedoxidative stress seen in this condition. Binding of Angioten-sin II to its type I receptor (AT1R) increases generation ofreactive oxygen species and we have shown an increase inAT1R in PE [2]. Therefore we investigated whether the RASis further augmented in PE at high altitudes.
Methods: Biopsies were collected, with informed, writtenconsent, from NT and PE women (ISSHP guidelines) at sea-level (UK) and high altitude, 3100 m (Leadville, Colorado)immediately after vaginal delivery or elective non-labouredCaesarean section. mRNA expression of the RAS compo-nents (angiotensinogen (AGT), prorenin, prorenin receptor,AT1R and angiotensin type 2 receptor) was determinedby qRT-PCR and normalised to 3 stably-expressed house-keeping genes using geNorm. Protein expression wassemi-quantitatively assessed by immunohistochemistry (Hscore) and compared between groups (Mann Whitney Utest).
Results: mRNA expression of all RAS components was unaf-fected by altitude in NT, but significantly lower for all in PE(P = 0.026 �< 0.0001). However, in NT, protein expressionof all components except AGT was significantly increasedat altitude (P = 0.037 �< 0.001). Protein expression of AGTand AT1R was also increased in altitude PE (P < 0.004 forboth).
Conclusion: We have shown the placental RAS to be acti-vated at altitude, with apparent differences in transcription/translation. Hypoxia-inducible transcription factors such asHIF-1 enhance transcription and translation of many genesencoding ‘‘rescue’’ proteins; this might explain the effects ofaltitude in NTs. HIF-1 is increased in PE, which might allowincreased protein expression at altitude in spite of lowermRNA.
Poster Presentations / Pregnancy Hypertension: An International Journal of Women’s Cardiovascular Health 3 (2013) 67–99 73
References
Kurlak LO et al. Abstract IFPA, Japan; 2012.Mistry HD et al. Placenta ];34:182–6.
doi:10.1016/j.preghy.2013.04.046
PP019. A new player in preeclampsia: The NF-E2-relatedfactor 2 (NRF2)Kweider Nisreen, Huppertz Berthold, Pecks Ulrich,Goecke Tamme, Pufe Thomas, Kadyrov Mamed, WruckChristoph Jan, Rath Werner
Introduction: Preeclampsia PE is characterized by dimin-ished antioxidant capacity. These enzymes are mainly regu-lated via the transcription factor Nrf2.
Objectives: PE is associated with an increase in Nrf2 activ-ity. Nrf2 involves also in the vascular homeostasis during PE.Respective hemodisturbances have been associated withimpaired invasion of the extravillous trophoblast EVT inearly onset IUGR associated with PE. To test this link, westudied in vitro the interaction between Nrf2 and VEGF, thentheir expression was determined in third trimester placentalbeds in cases of severe early onset IUGR/PE.
Methods: BeWo cells were used in the in vitro study.Wes-tern blot; ELISA and Dual Luciferase assay were applied. Full-thickness uterine tissues from 6 healthy and 6 women withsevere early onset IUGR/PE were used to study the expres-sion of VEGF, Nrf2 and 4-HNE in the EVT.
Results: Nrf2-activation and its downstream target pro-tein HO-1 augmented CO production, which in turn up-reg-ulated the expression of VEGF. EVT in cases with IUGR/PEshowed increased expression of Nrf2 and decreased VEGFintensity.
Conclusion: In early onset IUGR/PE the EVT experienceoxidative stress and try to counteract this by increasedexpression of Nrf2. However, since these cells fail to up-reg-ulate VEGF, Nrf2-activation does not occur, leading to fur-ther trophoblast damage. At the same time, in vitro datashow a protective role of the Nrf2/HO-1 pathway, whichmay have a therapeutic potential in PE.
doi:10.1016/j.preghy.2013.04.047
PP020. Evidence of a preventive role of Nrf2 inpreeclampsiaKweider Nisreen, Wruck Christoph Jan, Ludwig Andreas,Dreymueller Daniela, Goecke Tamme, Pecks Ulrich, PufeThomas, Rath Werner
Introduction: Smoking during pregnancy is associatedwith lower preeclampsia risk. This has been mainlyexplained through the effect of carbon monoxide CO.
Objectives: Recent studies showed that the activation ofheme oxygenase-1 HO-1 and consequently its metaboliteCO in cultured cells mediated an inhibition of sFlt-1 andsEng release, and an up-regulation of the endogenous VEGF.The transcriptional regulation of the HO-1 gene is majorlyregulated through the transcription factor Nrf2. The aim of
this study was to investigate in vitro the effect of HO-1-acti-vation via Nrf2 on the pro- and anti-angiogenic factors.
Methods: BeWo cells and HUVECs endothelial cells wereused to study the angiogenic effect of Nrf2-activation. ELISA,scratch and tube formation assay were mainly applied.
Results: The activation of HO-1 via Nrf2 lead to an increasein the protein levels of VEGF (control 64.75 pg/ml ± 4.3; Sul-foraphane-treated cells 128.2 pg/ml ± 6.5 p < 0.005) anddecrease in the augmented sFlt-1 in the supernatant of thetreated cells (control 186.3 pg/ml ± 28.7; H2O2-treatment2026 pg/ml ± 64, co-treatment with H2O2 and Sulforaphane1200 pg/ml ± 19.7 p < 0.01). Up-regulation of HO-1/COenhanced tube formation and migration of the endothelialcells.
Conclusion: The activation of HO-1/CO via Nrf2 inducersuch as sulforaphane inhibited in vitro the release of sFlt-1,thus the activation of Nrf2 during the first trimester mayimprove the balance of the pro- and anti-angiogenic factors.
doi:10.1016/j.preghy.2013.04.048
PP021. The role of the transcription factor Nrf2 in themurine placental developmentKweider Nisreen, Kistermann Jenny, Wruck ChristophJan, Pufe Thomas, Rath Werner
Introduction: The placenta is the key organ for successfulpregnancy and fetal growth. Oxidative stress during earlyhuman placental development is associated with pregnancy-related disorders. The transcription of many antioxidative-genes is mediated mainly through the transcription factorNrf2. Furthermore, a link between Nrf2, vascular homeostasisand extravillous trophoblast invasion has been discussed.
Objectives: Here, we investigated the placental phenotype,placental and fetal weight of the Nrf2 knockout (Nrf2�/�)and wild type (Nrf2+/+) mice and the vascular function ofthese placentas around embryonic day 18.5.
Methods: We performed H&E, Periodic Acid Schiff (PAS)and immunohistochemistry of paraffin-embedded mouseplacenta samples.
Results: There is no significant difference in both placentaland fetal weight of both geno types (Nrf2�/� and Nrf2+/+).Phenotypic analysis of ED 18.5 placentas showed presenceof trophoblast clusters in the labyrinth and frequentenlarged maternal blood lacunae. Furthermore, Nr2�/�
showed increased levels in the lipid peroxidation product4-hydroxinonoeal (4-HNE), which is a sensitive marker ofoxidative damage and lipid peroxidation.
Conclusion: This data point out the necessity of a func-tional Nrf2 for placental development, as it may interactwith the differentiation of the trophoblast lineage fromone side and to diminish the oxidative damage during preg-nancy from the other side.
doi:10.1016/j.preghy.2013.04.049
PP022. An animal model for eclampsiaLiu Lei, Liu Huishu, Huang Qian, Hu Bihui
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