Abstracts / Pregnancy Hypertension: An International Journal of Women’s Cardiovascular Health 2 (2012) 240–339
HO-1 and HIF-1 alpha mRNAs in placenta taken from preeclampsia and the effect of preeclampsia sera on their expression of choriocarcinoma cells and analysed the effect of placental hypoxia and serum factor on the expression of PlGF and sFlt-1 mRNA. Methods: Placenta and serum samples were taken from preeclampsia and normal pregnancy with informed consent. The choriocarcinoma cells (JEG-3) were cultured in 24-well tissue culture plate. The cells were cultured with preeclampsia and normal pregnant sera. The RNAs were purified from these cells 24 h after and placenta. The expressions of these mRNA were measured by using the real time PCR method (Applied Biosystems-7500). Results: The expression of PlGF mRNA decreased and that of sFlt-1mRNA increased in preeclampsia placenta. The expression of MTF-1 and HO-1 mRNA decreased. The correlation was found between the expression of PlGF and MTF-1 mRNA, PlGF and HO-1 mRNA and sFlt-1 and HO-1mRNA. Moreover, expression of sFlt-1mRNA increased and HO-1mRNA decreased in JEG-3 cells after incubation of preeclampsia sera. Conclusion: The changes of PlGFmRNA in preeclampsia placenta may relate to the expression of MTF-1 and HO-1 mRNA. The changes of sFlt-1mRNA may relate to the expression of HO-1 mRNA and serum factor. Not only hypoxia but also serum factor may play a role of the levels of PlGF and sFlt-1 in preeclampsia placenta. Disclosure of interest: None declared. doi:10.1016/j.preghy.2012.04.232
PP122. Lack of correlation between placental weight and circulating soluble fms-like tyrosine kinase-1 and placental growth factor suggests trophoblastic activity as a major contributor to the serum pool T. Wataganara *, B. Pratumvinit, J. Leetheerakul, S. Pongprasobchai, P. Lahfahroengron, J. Pooliam (Faculty of Medicine Siriraj Hospital, Bangkok, Thailand) Introduction: Serum levels of soluble fms-like tyrosine kinase-1 (sFlt-1) and placental growth factor (PlGF) are altered in preeclampsia and fetal growth restriction. Whether the alterations result from mere variation in
305
trophoblastic mass or trophoblastic activity in these diseases is still elusive. Objectives: Using placental weight as a surrogate for trophoblastic mass, we explored the relationship between placental weight at delivery and serum levels of sFlt-1 and PlGF at various gestational ages in normotensive pregnant Thai women. Methods: One hundred and forty two serum samples from healthy, normotensive, singleton Thai pregnant women were prospectively collected from 6 gestational age intervals; 10–14, 15–19, 20–24, 25–29, 30–34, and 35–40 weeks’. Analysis for the levels of sFlt-1 and PlGF was made from fresh specimens, using a commercially available automated system. Placental weight at delivery was followed in participants who remained unaffected with preeclampsia or fetal growth restriction until the time of delivery. Results: Statistical analyses were achieved from 140 participants. Quartiles for these markers were calculated for each gestational age interval. Serum sFlt-1 levels continuously increased through the pregnancy. Serum PlGF levels reached its peak levels at menstrual period of 25–29 weeks’, and then declined. Serum sFlt-1/PlGF ratios were consistently lower than that of the European population throughout the pregnancy. No significant correlation is found between placental weight at delivery, and serum levels of either sFlt-1 or PlGF at any gestational age interval. Conclusion: Ethnic variation of serum angiogenic markers is shown in this study. Lack of correlation between placental weight and serum angiogenic markers in normotensive individuals suggest that trophoblastic activity, and not number of the trophoblast, contribute to the alterations in circulating pool. This may facilitate a better understanding for its potential clinical applications of these biomarkers for other placental related diseases. Disclosure of interest: None declared doi:10.1016/j.preghy.2012.04.233
PP123. Evaluation of glycosaminoglycans in placental of pregnant women with and without preeclampsia N. Sass 1,*, E.A. Famá 2, M.A. Pinhal 3, T.A. Facca 1, R.S. Souza 3 (1 Obstetrics, Federal University of São Paulo,
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Abstracts / Pregnancy Hypertension: An International Journal of Women’s Cardiovascular Health 2 (2012) 240–339
São Paulo, Brazil, 2 Obstetrics, Federal University of São Paulo, São Paulo, Brazil, 3 Department of Biochemistry, ABC Medical School, São Paulo, Brazil) Introduction: Preeclampsia (PE) is a major cause of maternal and perinatal mobility and mortality and its etiology is not yet completely understood. Recently studies have shown the association between increased expression of glycosaminoglycans (GAGs) in placental of women with PE and its physiopathology. Objectives: Identify and quantify GAGs in placental of pregnant women in healthy pregnancy and PE. Methods: Case-control study with 44 patients, control group (CG) n = 29 and n = 15 PE group. All patients were submitted to placental tissue resection (sample size of 5 5 cm with the umbilical cord insertion in the center). The tissue was conserved in acetone. The GAGs’ analysis consisted of centrifugation, proteolysis, precipitation, and electrophoresis Results: Average age and gestational age in CG and PE were 27.33 years and 39.02 weeks and 24.17 and 36.90, respectively. In CG 68.96% (20/29) were Caucasian and 80.00% (12/15) in PE. We found in CG 34.48% (10/29) of primiparous and 40.00% (6/15) in PE. The average 24-hour proteinuria in PE was 554.28 g/24hs. The average birth weight was 3333.31 g in CG and 2972.66 in PE. The mean ± standard deviation of dermatan sulfate (DS), heparan sulfate (HS) and hyaluronic acid (HA) in CG and PE were: 0,100 lg/mg of tissue ±0,005 and 0,144 ±0,071; 0,077 ± 0,041 and 0,113 ± 0,061; 1,281 ± 1,857 and 3,076 ± 4,930. Conclusion: GAGs are increased in PE when compared to normal pregnancy. The expression of HA and HS was twice higher in PE. More studies are needed to determine the correlation between GAGs and physiopathology of PE. (see Table 1) Disclosure of interest: None declared. Table 1 Mean and standard deviation of gilcosaminoglicanos expression in placental of healthy pregnancy and preeclampsia. DS (lg/mg tissue)
HS (lg/mg tissue)
HA (lg/mg tissue)
Control Group 0.100 ± 0.055 0.077 ± 0.041 1.281 ± 1.857 PE Group 0.144 ± 0.071 0.113 ± 0.061 3.076 ± 4.930 DS: dermatan sulfate, HS: heparan sulfate, HA: hyaluronic acid
doi:10.1016/j.preghy.2012.04.234
PP124. Trofoblastic invasion: The role of subunits STAT1 and STAT3 H.M.F. Marconato 1, V.J.A. Panico 1, L. Saragiotto 1, T.L. Machado 1, I.R.C. Lotti 1, B.M. Kabbach 1, J.M. Garcia 1, F.L.P. Sousa 1,2,*, D.M. Morales Prieto 2, N. Sass 3, U.R. Markert 2 (1 Obstetrics, UNILUS – Lusíada Foundation, Faculty of Medical Sciences of Santos, Santos, Brazil, 2 Obstetrics, Placenta-Laboratories, University Hospital Jena, Jena, Germany, 3 Obstetrics, Federal University of São Paulo, São Paulo, Brazil)
Introduction: The trophoblastic migration/invasion are controlled by cytokines and growth factors that use intracellular pathways of signal to promote the regulation of gene expression, proliferation, cells differentiation, angiogenesis and embryonic development. The most important mediator of cytokine in trophoblastic invasion is the Janus-Kinase/signal transducer and activator of transcription (JAK/STAT). STATs are amino acids, compounds of 700–850 variable long-chain with isoforms a and b and molecular weight between 83–113 kDa. The role of these factors in the pregnancy set up may contribute to adopt interventions that could contribute to prophylaxis and/or treatment of abnormalities in the course of gestation when installed early. Objectives: Search on database the role of STAT in the process of trophoblastic invasion with emphasis on subunits STAT1 and STAT3. Methods: This is a review performed on PubMed database. Have been included Studies found from 1992 (the year of discovery of STATs) until July 2011, without language restriction. The descriptors were: ‘‘Signal transducers and activators of transcription ‘‘and’’ Trophoblast ’’. In the end we excluded bibliographical review. Results: Five of the six selected papers studied the role of STAT3 in the physiology of the trophoblastc invasion process. One of them, indirectly by selection process of lactobacilli of vaginal flora endogenous, during change of vaginal pH on pregnancy, altering the release of greater or lesser number of Interleukin-10 which modulates the activation JAK/ STAT. Among them, one of the study refers to involvement of STAT1 in the immunomodulation of interface fetusmother. Conclusion: STAT3 is directly involved in the process of trophoblast invasion either in its endometrium adherence to, angiogenesis, invasion and regulation of invasion. And STAT1 is involved in immunomodulation through its suppression by trophoblast STAT utron. Several soluble factors that are generally present in the decidua, especially hepatocyte growth factor, granulocyte macrophagocytic-colony stimulating factors, interleukin-6, interleukin-11 and inhibition leukemia factor , which have been described by using the JAK-STAT activating STAT1 and STAT3 for intracellular signaling and from this process may influence the invasion trophoblast. Disclosure of interest: None declared doi:10.1016/j.preghy.2012.04.235
PP125. High mobility group box 1 protein (HMGB1): A pathogenic role in preeclampsia? P.-A. Pradervand 1,*, B. Waeber 2, L. Liaudet 3, Y. Vial 1, F. Feihl 2 (1 Département de Gynécologie-Obstétrique, Switzerland, 2 Division de Physiopathologie clinique, Switzerland, 3 Service de Médecine intensive adulte, CHUV, Lausanne, Switzerland) Introduction: A central pathogenic mechanism of preeclampsia is believed to be the production by the hypoxic
HO-1 and HIF-1 alpha mRNAs in placenta taken from preeclampsia and the effect of preeclampsia sera on their expression of choriocarcinoma cells and analysed the effect of placental hypoxia and serum factor on the expression of PlGF and sFlt-1 mRNA. Methods: Placenta and serum samples were taken from preeclampsia and normal pregnancy with informed consent. The choriocarcinoma cells (JEG-3) were cultured in 24-well tissue culture plate. The cells were cultured with preeclampsia and normal pregnant sera. The RNAs were purified from these cells 24 h after and placenta. The expressions of these mRNA were measured by using the real time PCR method (Applied Biosystems-7500). Results: The expression of PlGF mRNA decreased and that of sFlt-1mRNA increased in preeclampsia placenta. The expression of MTF-1 and HO-1 mRNA decreased. The correlation was found between the expression of PlGF and MTF-1 mRNA, PlGF and HO-1 mRNA and sFlt-1 and HO-1mRNA. Moreover, expression of sFlt-1mRNA increased and HO-1mRNA decreased in JEG-3 cells after incubation of preeclampsia sera. Conclusion: The changes of PlGFmRNA in preeclampsia placenta may relate to the expression of MTF-1 and HO-1 mRNA. The changes of sFlt-1mRNA may relate to the expression of HO-1 mRNA and serum factor. Not only hypoxia but also serum factor may play a role of the levels of PlGF and sFlt-1 in preeclampsia placenta. Disclosure of interest: None declared. doi:10.1016/j.preghy.2012.04.232
PP122. Lack of correlation between placental weight and circulating soluble fms-like tyrosine kinase-1 and placental growth factor suggests trophoblastic activity as a major contributor to the serum pool T. Wataganara *, B. Pratumvinit, J. Leetheerakul, S. Pongprasobchai, P. Lahfahroengron, J. Pooliam (Faculty of Medicine Siriraj Hospital, Bangkok, Thailand) Introduction: Serum levels of soluble fms-like tyrosine kinase-1 (sFlt-1) and placental growth factor (PlGF) are altered in preeclampsia and fetal growth restriction. Whether the alterations result from mere variation in
305
trophoblastic mass or trophoblastic activity in these diseases is still elusive. Objectives: Using placental weight as a surrogate for trophoblastic mass, we explored the relationship between placental weight at delivery and serum levels of sFlt-1 and PlGF at various gestational ages in normotensive pregnant Thai women. Methods: One hundred and forty two serum samples from healthy, normotensive, singleton Thai pregnant women were prospectively collected from 6 gestational age intervals; 10–14, 15–19, 20–24, 25–29, 30–34, and 35–40 weeks’. Analysis for the levels of sFlt-1 and PlGF was made from fresh specimens, using a commercially available automated system. Placental weight at delivery was followed in participants who remained unaffected with preeclampsia or fetal growth restriction until the time of delivery. Results: Statistical analyses were achieved from 140 participants. Quartiles for these markers were calculated for each gestational age interval. Serum sFlt-1 levels continuously increased through the pregnancy. Serum PlGF levels reached its peak levels at menstrual period of 25–29 weeks’, and then declined. Serum sFlt-1/PlGF ratios were consistently lower than that of the European population throughout the pregnancy. No significant correlation is found between placental weight at delivery, and serum levels of either sFlt-1 or PlGF at any gestational age interval. Conclusion: Ethnic variation of serum angiogenic markers is shown in this study. Lack of correlation between placental weight and serum angiogenic markers in normotensive individuals suggest that trophoblastic activity, and not number of the trophoblast, contribute to the alterations in circulating pool. This may facilitate a better understanding for its potential clinical applications of these biomarkers for other placental related diseases. Disclosure of interest: None declared doi:10.1016/j.preghy.2012.04.233
PP123. Evaluation of glycosaminoglycans in placental of pregnant women with and without preeclampsia N. Sass 1,*, E.A. Famá 2, M.A. Pinhal 3, T.A. Facca 1, R.S. Souza 3 (1 Obstetrics, Federal University of São Paulo,
306
Abstracts / Pregnancy Hypertension: An International Journal of Women’s Cardiovascular Health 2 (2012) 240–339
São Paulo, Brazil, 2 Obstetrics, Federal University of São Paulo, São Paulo, Brazil, 3 Department of Biochemistry, ABC Medical School, São Paulo, Brazil) Introduction: Preeclampsia (PE) is a major cause of maternal and perinatal mobility and mortality and its etiology is not yet completely understood. Recently studies have shown the association between increased expression of glycosaminoglycans (GAGs) in placental of women with PE and its physiopathology. Objectives: Identify and quantify GAGs in placental of pregnant women in healthy pregnancy and PE. Methods: Case-control study with 44 patients, control group (CG) n = 29 and n = 15 PE group. All patients were submitted to placental tissue resection (sample size of 5 5 cm with the umbilical cord insertion in the center). The tissue was conserved in acetone. The GAGs’ analysis consisted of centrifugation, proteolysis, precipitation, and electrophoresis Results: Average age and gestational age in CG and PE were 27.33 years and 39.02 weeks and 24.17 and 36.90, respectively. In CG 68.96% (20/29) were Caucasian and 80.00% (12/15) in PE. We found in CG 34.48% (10/29) of primiparous and 40.00% (6/15) in PE. The average 24-hour proteinuria in PE was 554.28 g/24hs. The average birth weight was 3333.31 g in CG and 2972.66 in PE. The mean ± standard deviation of dermatan sulfate (DS), heparan sulfate (HS) and hyaluronic acid (HA) in CG and PE were: 0,100 lg/mg of tissue ±0,005 and 0,144 ±0,071; 0,077 ± 0,041 and 0,113 ± 0,061; 1,281 ± 1,857 and 3,076 ± 4,930. Conclusion: GAGs are increased in PE when compared to normal pregnancy. The expression of HA and HS was twice higher in PE. More studies are needed to determine the correlation between GAGs and physiopathology of PE. (see Table 1) Disclosure of interest: None declared. Table 1 Mean and standard deviation of gilcosaminoglicanos expression in placental of healthy pregnancy and preeclampsia. DS (lg/mg tissue)
HS (lg/mg tissue)
HA (lg/mg tissue)
Control Group 0.100 ± 0.055 0.077 ± 0.041 1.281 ± 1.857 PE Group 0.144 ± 0.071 0.113 ± 0.061 3.076 ± 4.930 DS: dermatan sulfate, HS: heparan sulfate, HA: hyaluronic acid
doi:10.1016/j.preghy.2012.04.234
PP124. Trofoblastic invasion: The role of subunits STAT1 and STAT3 H.M.F. Marconato 1, V.J.A. Panico 1, L. Saragiotto 1, T.L. Machado 1, I.R.C. Lotti 1, B.M. Kabbach 1, J.M. Garcia 1, F.L.P. Sousa 1,2,*, D.M. Morales Prieto 2, N. Sass 3, U.R. Markert 2 (1 Obstetrics, UNILUS – Lusíada Foundation, Faculty of Medical Sciences of Santos, Santos, Brazil, 2 Obstetrics, Placenta-Laboratories, University Hospital Jena, Jena, Germany, 3 Obstetrics, Federal University of São Paulo, São Paulo, Brazil)
Introduction: The trophoblastic migration/invasion are controlled by cytokines and growth factors that use intracellular pathways of signal to promote the regulation of gene expression, proliferation, cells differentiation, angiogenesis and embryonic development. The most important mediator of cytokine in trophoblastic invasion is the Janus-Kinase/signal transducer and activator of transcription (JAK/STAT). STATs are amino acids, compounds of 700–850 variable long-chain with isoforms a and b and molecular weight between 83–113 kDa. The role of these factors in the pregnancy set up may contribute to adopt interventions that could contribute to prophylaxis and/or treatment of abnormalities in the course of gestation when installed early. Objectives: Search on database the role of STAT in the process of trophoblastic invasion with emphasis on subunits STAT1 and STAT3. Methods: This is a review performed on PubMed database. Have been included Studies found from 1992 (the year of discovery of STATs) until July 2011, without language restriction. The descriptors were: ‘‘Signal transducers and activators of transcription ‘‘and’’ Trophoblast ’’. In the end we excluded bibliographical review. Results: Five of the six selected papers studied the role of STAT3 in the physiology of the trophoblastc invasion process. One of them, indirectly by selection process of lactobacilli of vaginal flora endogenous, during change of vaginal pH on pregnancy, altering the release of greater or lesser number of Interleukin-10 which modulates the activation JAK/ STAT. Among them, one of the study refers to involvement of STAT1 in the immunomodulation of interface fetusmother. Conclusion: STAT3 is directly involved in the process of trophoblast invasion either in its endometrium adherence to, angiogenesis, invasion and regulation of invasion. And STAT1 is involved in immunomodulation through its suppression by trophoblast STAT utron. Several soluble factors that are generally present in the decidua, especially hepatocyte growth factor, granulocyte macrophagocytic-colony stimulating factors, interleukin-6, interleukin-11 and inhibition leukemia factor , which have been described by using the JAK-STAT activating STAT1 and STAT3 for intracellular signaling and from this process may influence the invasion trophoblast. Disclosure of interest: None declared doi:10.1016/j.preghy.2012.04.235
PP125. High mobility group box 1 protein (HMGB1): A pathogenic role in preeclampsia? P.-A. Pradervand 1,*, B. Waeber 2, L. Liaudet 3, Y. Vial 1, F. Feihl 2 (1 Département de Gynécologie-Obstétrique, Switzerland, 2 Division de Physiopathologie clinique, Switzerland, 3 Service de Médecine intensive adulte, CHUV, Lausanne, Switzerland) Introduction: A central pathogenic mechanism of preeclampsia is believed to be the production by the hypoxic
