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PP007. Nicotinic acetylcholine receptor subunits in the pre-eclamptic and cigarette smoke exposed human placenta
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Poster Presentations / Pregnancy Hypertension: An International Journal of Women’s Cardiovascular Health 3 (2013) 67–99
PP007. Nicotinic acetylcholine receptor subunits in the pre-eclamptic and cigarette smoke exposed human placenta Ghazavi Emma, David Rona, Hinton Tina, Makris Angela, Hennessy Annemarie, Machaalani Rita Introduction: Cigarette smoking during pregnancy is associated with low birth weight, prematurity and neonatal morbidity and mortality. Nicotine, a major constituent of cigarette smoke, binds to nicotinic acetylcholine receptors (nAChRs). To date, 16 mammalian nAChR subunits have been identified. The effect of smoking on these subunits in human placenta has not yet been determined. Smoking is also associated with reduced pre-eclampsia (PE) risk and its protective effects may occur via changes in nAChRs. Objectives: This study aimed to determine which nAChR subunits are present in the normal human placenta, and whether any changes are occur from smoking or PE. Methods: Using RT-qPCR, all 16 nAChR subunits were investigated in normal, healthy human placentas, and mRNA expression compared between controls (n = 8), smokers (n = 8) and PE (n = 7). Results All 16 nAChR subunits were expressed in healthy placentas. Smoke exposure significantly increased a2 (p = 0.006) and a9 (p = 0.038), and decreased d (p = 0.013), subunit mRNA expression. In PE placentas, b1 (p = 0.048) and b2 (p = 0.031) subunit mRNA expression was increased. Conclusion: Nicotine exposure in pregnancy increases nAChR subunit mRNAs that play a role in vasoconstriction and amino acid uptake, possibly contributing to abnormalities in placentas from smoking mothers. Different subunits were affected in PE placentas, thus the hypoxic environment may induce changes in endogenous ACh levels, yielding compensatory increases in b1 and b2 subunits. doi:10.1016/j.preghy.2013.04.035
PP008. Placental klotho gene in preeclampsia Giannubilo Stefano Raffaele, Cecati Monia, Saccucci Franca, Emanuelli Monica, Tranquilli Andrea Luigi Introduction: An aging-suppressor gene, klotho, is a candidate factor for vascular disease because its deficiency leads to impaired endothelium-dependent vasodilation and impaired angiogenesis. This protein is involved in several metabolic pathways such as the insulin-like growth factor 1 (IGF-1), apoptosis, angiotensin-II-induced events in the kidney and oxidative stress. Objectives: The aim of this study was to determine the difference of klotho genotiping and expression in the placentas of women with normal and preeclamptic pregnancies. Method/Design: Placental tissue was collected from normal pregnancies (n = 12) and pregnancies complicated by Preeclampsia (n = 12), matched for gestational age. Klotho genotyping and expression was determined using real-time quantitative polymerase chain reaction (PCR) and Western blot, respectively. Results: A polymorphism for 744 G/A mutation was significantly more common in the pathological group, with an odds ratio (OR) of 3.00 (1.02–8.81; 95% CI). The expression
levels of both klotho isoforms and of the short klotho isoform were lower (80%) in the Preeclampsia group as compared to matched controls. Results of Western Blot agreed with those from Real-Time PCR. Conclusion: In preeclamptic pregnancies there are a genotyping polymorphism and a reduced expression of klotho gene. Given its role in cardiovascular disease in aging, it may link preeclamptic mothers and their offsprings to long term cardiovascular outcomes. doi:10.1016/j.preghy.2013.04.036
PP009. Hypoxia alters syncytiotrophoblastic microparticles (STBM)-related coagulation capacities Göhner Claudia, Schlembach Dietmar, Schleussner Ekkehard, Markert Udo R., Fitzgerald Justine S. Introduction: Endothelial dysfunction is thought to be the cause of preeclampsia (PE) symptoms. Severe forms of PE are correlated to the release of syncytiotrophoblastic microparticles (STBM), which triggers inflammatory processes on the endothelium. The thrombogenic potential of STBMs is not well characterized. We investigated the pro-coagulant activity of STBMs. Methods: STBMs were derived from placentae perfused under norm- and hypoxic conditions and quantified with our house-‘‘ELISA’’. Surface phospholipid-dependent thrombin formation of microparticles was determined with a commercial ELISA. Rate and absolute platelet aggregation in platelet-rich plasma (PRP), while fibrin production in platelet free plasma, was measured in absence and presence of STBMs using a PAP-4 aggregometer. Results: STBM concentration and STBM-induced thrombin formation is stable under normoxic and elevated under hypoxic conditions. STBMs derived from hypoxic placentae increase the rate of fibrinogenesis. Maximum platelet aggregation is stable under normoxic, while highly variable under hypoxic conditions. STBMs of hypoxic placentae cannot alter the rate of platelet aggregation, while normoxic STBMs adjust the rate according to need. Conclusion: Hypoxia negatively affects fibrinogenesis and the platelet aggregation-modulating effects of STBMs, which might be due to a phenotype alteration. All observed results may contribute to the impaired microcirculation seen in PE. doi:10.1016/j.preghy.2013.04.037
PP010. Alpha-1-microglobulin protects from heme induced placenta and kidney damage in a pregnant ewe model for preeclampsia Edström-Hägerwall Anneli, Hansson Stefan, Casslén Vera, Rosenlöf Lena, Axelsson Josefin, Rippe Bengt, Marsal Karel, Mörgelin Matthias, Åkerström Bo, Gram Magnus Introduction: Previous gene expression analysis have identified fetal hemoglobin (HbF) as a plausible etiological factor in preeclampsia. Free hemoglobin and its degradation products, e.g. heme, are known to cause oxidative stress,
Poster Presentations / Pregnancy Hypertension: An International Journal of Women’s Cardiovascular Health 3 (2013) 67–99
PP007. Nicotinic acetylcholine receptor subunits in the pre-eclamptic and cigarette smoke exposed human placenta Ghazavi Emma, David Rona, Hinton Tina, Makris Angela, Hennessy Annemarie, Machaalani Rita Introduction: Cigarette smoking during pregnancy is associated with low birth weight, prematurity and neonatal morbidity and mortality. Nicotine, a major constituent of cigarette smoke, binds to nicotinic acetylcholine receptors (nAChRs). To date, 16 mammalian nAChR subunits have been identified. The effect of smoking on these subunits in human placenta has not yet been determined. Smoking is also associated with reduced pre-eclampsia (PE) risk and its protective effects may occur via changes in nAChRs. Objectives: This study aimed to determine which nAChR subunits are present in the normal human placenta, and whether any changes are occur from smoking or PE. Methods: Using RT-qPCR, all 16 nAChR subunits were investigated in normal, healthy human placentas, and mRNA expression compared between controls (n = 8), smokers (n = 8) and PE (n = 7). Results All 16 nAChR subunits were expressed in healthy placentas. Smoke exposure significantly increased a2 (p = 0.006) and a9 (p = 0.038), and decreased d (p = 0.013), subunit mRNA expression. In PE placentas, b1 (p = 0.048) and b2 (p = 0.031) subunit mRNA expression was increased. Conclusion: Nicotine exposure in pregnancy increases nAChR subunit mRNAs that play a role in vasoconstriction and amino acid uptake, possibly contributing to abnormalities in placentas from smoking mothers. Different subunits were affected in PE placentas, thus the hypoxic environment may induce changes in endogenous ACh levels, yielding compensatory increases in b1 and b2 subunits. doi:10.1016/j.preghy.2013.04.035
PP008. Placental klotho gene in preeclampsia Giannubilo Stefano Raffaele, Cecati Monia, Saccucci Franca, Emanuelli Monica, Tranquilli Andrea Luigi Introduction: An aging-suppressor gene, klotho, is a candidate factor for vascular disease because its deficiency leads to impaired endothelium-dependent vasodilation and impaired angiogenesis. This protein is involved in several metabolic pathways such as the insulin-like growth factor 1 (IGF-1), apoptosis, angiotensin-II-induced events in the kidney and oxidative stress. Objectives: The aim of this study was to determine the difference of klotho genotiping and expression in the placentas of women with normal and preeclamptic pregnancies. Method/Design: Placental tissue was collected from normal pregnancies (n = 12) and pregnancies complicated by Preeclampsia (n = 12), matched for gestational age. Klotho genotyping and expression was determined using real-time quantitative polymerase chain reaction (PCR) and Western blot, respectively. Results: A polymorphism for 744 G/A mutation was significantly more common in the pathological group, with an odds ratio (OR) of 3.00 (1.02–8.81; 95% CI). The expression
levels of both klotho isoforms and of the short klotho isoform were lower (80%) in the Preeclampsia group as compared to matched controls. Results of Western Blot agreed with those from Real-Time PCR. Conclusion: In preeclamptic pregnancies there are a genotyping polymorphism and a reduced expression of klotho gene. Given its role in cardiovascular disease in aging, it may link preeclamptic mothers and their offsprings to long term cardiovascular outcomes. doi:10.1016/j.preghy.2013.04.036
PP009. Hypoxia alters syncytiotrophoblastic microparticles (STBM)-related coagulation capacities Göhner Claudia, Schlembach Dietmar, Schleussner Ekkehard, Markert Udo R., Fitzgerald Justine S. Introduction: Endothelial dysfunction is thought to be the cause of preeclampsia (PE) symptoms. Severe forms of PE are correlated to the release of syncytiotrophoblastic microparticles (STBM), which triggers inflammatory processes on the endothelium. The thrombogenic potential of STBMs is not well characterized. We investigated the pro-coagulant activity of STBMs. Methods: STBMs were derived from placentae perfused under norm- and hypoxic conditions and quantified with our house-‘‘ELISA’’. Surface phospholipid-dependent thrombin formation of microparticles was determined with a commercial ELISA. Rate and absolute platelet aggregation in platelet-rich plasma (PRP), while fibrin production in platelet free plasma, was measured in absence and presence of STBMs using a PAP-4 aggregometer. Results: STBM concentration and STBM-induced thrombin formation is stable under normoxic and elevated under hypoxic conditions. STBMs derived from hypoxic placentae increase the rate of fibrinogenesis. Maximum platelet aggregation is stable under normoxic, while highly variable under hypoxic conditions. STBMs of hypoxic placentae cannot alter the rate of platelet aggregation, while normoxic STBMs adjust the rate according to need. Conclusion: Hypoxia negatively affects fibrinogenesis and the platelet aggregation-modulating effects of STBMs, which might be due to a phenotype alteration. All observed results may contribute to the impaired microcirculation seen in PE. doi:10.1016/j.preghy.2013.04.037
PP010. Alpha-1-microglobulin protects from heme induced placenta and kidney damage in a pregnant ewe model for preeclampsia Edström-Hägerwall Anneli, Hansson Stefan, Casslén Vera, Rosenlöf Lena, Axelsson Josefin, Rippe Bengt, Marsal Karel, Mörgelin Matthias, Åkerström Bo, Gram Magnus Introduction: Previous gene expression analysis have identified fetal hemoglobin (HbF) as a plausible etiological factor in preeclampsia. Free hemoglobin and its degradation products, e.g. heme, are known to cause oxidative stress,