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PP020. Evidence of a preventive role of Nrf2 in preeclampsia
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Poster Presentations / Pregnancy Hypertension: An International Journal of Women’s Cardiovascular Health 3 (2013) 67–99
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-related factor 2 (NRF2) Kweider Nisreen, Huppertz Berthold, Pecks Ulrich, Goecke Tamme, Pufe Thomas, Kadyrov Mamed, Wruck Christoph Jan, Rath Werner Introduction: Preeclampsia PE is characterized by diminished antioxidant capacity. These enzymes are mainly regulated via the transcription factor Nrf2. Objectives: PE is associated with an increase in Nrf2 activity. Nrf2 involves also in the vascular homeostasis during PE. Respective hemodisturbances have been associated with impaired invasion of the extravillous trophoblast EVT in early onset IUGR associated with PE. To test this link, we studied in vitro the interaction between Nrf2 and VEGF, then their expression was determined in third trimester placental beds in cases of severe early onset IUGR/PE. Methods: BeWo cells were used in the in vitro study.Western blot; ELISA and Dual Luciferase assay were applied. Fullthickness uterine tissues from 6 healthy and 6 women with severe early onset IUGR/PE were used to study the expression of VEGF, Nrf2 and 4-HNE in the EVT. Results: Nrf2-activation and its downstream target protein HO-1 augmented CO production, which in turn up-regulated the expression of VEGF. EVT in cases with IUGR/PE showed increased expression of Nrf2 and decreased VEGF intensity. Conclusion: In early onset IUGR/PE the EVT experience oxidative stress and try to counteract this by increased expression of Nrf2. However, since these cells fail to up-regulate VEGF, Nrf2-activation does not occur, leading to further trophoblast damage. At the same time, in vitro data show a protective role of the Nrf2/HO-1 pathway, which may have a therapeutic potential in PE. doi:10.1016/j.preghy.2013.04.047
PP020. Evidence of a preventive role of Nrf2 in preeclampsia Kweider Nisreen, Wruck Christoph Jan, Ludwig Andreas, Dreymueller Daniela, Goecke Tamme, Pecks Ulrich, Pufe Thomas, Rath Werner Introduction: Smoking during pregnancy is associated with lower preeclampsia risk. This has been mainly explained through the effect of carbon monoxide CO. Objectives: Recent studies showed that the activation of heme oxygenase-1 HO-1 and consequently its metabolite CO in cultured cells mediated an inhibition of sFlt-1 and sEng release, and an up-regulation of the endogenous VEGF. The transcriptional regulation of the HO-1 gene is majorly regulated through the transcription factor Nrf2. The aim of
this study was to investigate in vitro the effect of HO-1-activation via Nrf2 on the pro- and anti-angiogenic factors. Methods: BeWo cells and HUVECs endothelial cells were used 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 increase in the protein levels of VEGF (control 64.75 pg/ml ± 4.3; Sulforaphane-treated cells 128.2 pg/ml ± 6.5 p < 0.005) and decrease in the augmented sFlt-1 in the supernatant of the treated cells (control 186.3 pg/ml ± 28.7; H2O2-treatment 2026 pg/ml ± 64, co-treatment with H2O2 and Sulforaphane 1200 pg/ml ± 19.7 p < 0.01). Up-regulation of HO-1/CO enhanced tube formation and migration of the endothelial cells. Conclusion: The activation of HO-1/CO via Nrf2 inducer such as sulforaphane inhibited in vitro the release of sFlt-1, thus the activation of Nrf2 during the first trimester may improve 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 the murine placental development Kweider Nisreen, Kistermann Jenny, Wruck Christoph Jan, Pufe Thomas, Rath Werner Introduction: The placenta is the key organ for successful pregnancy and fetal growth. Oxidative stress during early human placental development is associated with pregnancyrelated disorders. The transcription of many antioxidativegenes is mediated mainly through the transcription factor Nrf2. Furthermore, a link between Nrf2, vascular homeostasis and 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 of these placentas around embryonic day 18.5. Methods: We performed H&E, Periodic Acid Schiff (PAS) and immunohistochemistry of paraffin-embedded mouse placenta samples. Results: There is no significant difference in both placental and fetal weight of both geno types (Nrf2 / and Nrf2+/+). Phenotypic analysis of ED 18.5 placentas showed presence of trophoblast clusters in the labyrinth and frequent enlarged maternal blood lacunae. Furthermore, Nr2 / showed increased levels in the lipid peroxidation product 4-hydroxinonoeal (4-HNE), which is a sensitive marker of oxidative damage and lipid peroxidation. Conclusion: This data point out the necessity of a functional Nrf2 for placental development, as it may interact with the differentiation of the trophoblast lineage from one side and to diminish the oxidative damage during pregnancy from the other side. doi:10.1016/j.preghy.2013.04.049
PP022. An animal model for eclampsia Liu Lei, Liu Huishu, Huang Qian, Hu Bihui
Poster Presentations / Pregnancy Hypertension: An International Journal of Women’s Cardiovascular Health 3 (2013) 67–99
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-related factor 2 (NRF2) Kweider Nisreen, Huppertz Berthold, Pecks Ulrich, Goecke Tamme, Pufe Thomas, Kadyrov Mamed, Wruck Christoph Jan, Rath Werner Introduction: Preeclampsia PE is characterized by diminished antioxidant capacity. These enzymes are mainly regulated via the transcription factor Nrf2. Objectives: PE is associated with an increase in Nrf2 activity. Nrf2 involves also in the vascular homeostasis during PE. Respective hemodisturbances have been associated with impaired invasion of the extravillous trophoblast EVT in early onset IUGR associated with PE. To test this link, we studied in vitro the interaction between Nrf2 and VEGF, then their expression was determined in third trimester placental beds in cases of severe early onset IUGR/PE. Methods: BeWo cells were used in the in vitro study.Western blot; ELISA and Dual Luciferase assay were applied. Fullthickness uterine tissues from 6 healthy and 6 women with severe early onset IUGR/PE were used to study the expression of VEGF, Nrf2 and 4-HNE in the EVT. Results: Nrf2-activation and its downstream target protein HO-1 augmented CO production, which in turn up-regulated the expression of VEGF. EVT in cases with IUGR/PE showed increased expression of Nrf2 and decreased VEGF intensity. Conclusion: In early onset IUGR/PE the EVT experience oxidative stress and try to counteract this by increased expression of Nrf2. However, since these cells fail to up-regulate VEGF, Nrf2-activation does not occur, leading to further trophoblast damage. At the same time, in vitro data show a protective role of the Nrf2/HO-1 pathway, which may have a therapeutic potential in PE. doi:10.1016/j.preghy.2013.04.047
PP020. Evidence of a preventive role of Nrf2 in preeclampsia Kweider Nisreen, Wruck Christoph Jan, Ludwig Andreas, Dreymueller Daniela, Goecke Tamme, Pecks Ulrich, Pufe Thomas, Rath Werner Introduction: Smoking during pregnancy is associated with lower preeclampsia risk. This has been mainly explained through the effect of carbon monoxide CO. Objectives: Recent studies showed that the activation of heme oxygenase-1 HO-1 and consequently its metabolite CO in cultured cells mediated an inhibition of sFlt-1 and sEng release, and an up-regulation of the endogenous VEGF. The transcriptional regulation of the HO-1 gene is majorly regulated through the transcription factor Nrf2. The aim of
this study was to investigate in vitro the effect of HO-1-activation via Nrf2 on the pro- and anti-angiogenic factors. Methods: BeWo cells and HUVECs endothelial cells were used 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 increase in the protein levels of VEGF (control 64.75 pg/ml ± 4.3; Sulforaphane-treated cells 128.2 pg/ml ± 6.5 p < 0.005) and decrease in the augmented sFlt-1 in the supernatant of the treated cells (control 186.3 pg/ml ± 28.7; H2O2-treatment 2026 pg/ml ± 64, co-treatment with H2O2 and Sulforaphane 1200 pg/ml ± 19.7 p < 0.01). Up-regulation of HO-1/CO enhanced tube formation and migration of the endothelial cells. Conclusion: The activation of HO-1/CO via Nrf2 inducer such as sulforaphane inhibited in vitro the release of sFlt-1, thus the activation of Nrf2 during the first trimester may improve 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 the murine placental development Kweider Nisreen, Kistermann Jenny, Wruck Christoph Jan, Pufe Thomas, Rath Werner Introduction: The placenta is the key organ for successful pregnancy and fetal growth. Oxidative stress during early human placental development is associated with pregnancyrelated disorders. The transcription of many antioxidativegenes is mediated mainly through the transcription factor Nrf2. Furthermore, a link between Nrf2, vascular homeostasis and 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 of these placentas around embryonic day 18.5. Methods: We performed H&E, Periodic Acid Schiff (PAS) and immunohistochemistry of paraffin-embedded mouse placenta samples. Results: There is no significant difference in both placental and fetal weight of both geno types (Nrf2 / and Nrf2+/+). Phenotypic analysis of ED 18.5 placentas showed presence of trophoblast clusters in the labyrinth and frequent enlarged maternal blood lacunae. Furthermore, Nr2 / showed increased levels in the lipid peroxidation product 4-hydroxinonoeal (4-HNE), which is a sensitive marker of oxidative damage and lipid peroxidation. Conclusion: This data point out the necessity of a functional Nrf2 for placental development, as it may interact with the differentiation of the trophoblast lineage from one side and to diminish the oxidative damage during pregnancy from the other side. doi:10.1016/j.preghy.2013.04.049
PP022. An animal model for eclampsia Liu Lei, Liu Huishu, Huang Qian, Hu Bihui