- Email: [email protected]
6J mice
210
Abstracts / Pregnancy Hypertension: An International Journal of Women’s Cardiovascular Health 6 (2016) 178–252
Preeclampsia courses with endothelial and insulin resistance. Insulin dilates umbilical vein requiring A2A adenosine receptor (A2AAR) activation in normal pregnancies; however, whether A2AAR are involved in late-onset preeclampsia (LOPE)-reduced insulin dilation is unknown. LOPE increases maternal and foetal plasma adenosine and A2BAR expression in human umbilical vein endothelial cells (HUVECs). A2BAR involvement and insulin effect on the foetoplacental vascular function in LOPE is unknown. Aim: To evaluate A2AAR and A2BAR involvement on insulin effect on endothelial function in umbilical veins and HUVECs from LOPE. Methods: Protein abundance (total and phosphorylated) of p44/42mapk, protein kinase B/Akt (Akt), endothelial nitric oxide synthase (eNOS) were detected by Western blot. Assays were in the absence or presence (8 h) of 1 nM insulin, 30 nM CGS21680 (A2AAR agonist), 10 nM ZM241385 (A2AAR antagonist), 0.1 nM BAY606583 (A2BAR agonist), or 30 nM MRS1754 (A2BAR antagonist). Vascular response to insulin (0.1–1000 nM, 5 min) was measured in preconstricted umbilical vein rings (wire myography) in the absence or presence of adenosine (1 mM) and/or A2AAR and A2BAR antagonists. L-Citrulline level was measured by HPLC in the absence or presence (8 h) of NG-nitro- L-arginine methyl ester (100 lM) in HUVECs. Results: Insulin increased Akt (1.3 0.1 fold), p44/42mapk (1.2 0.1 fold), Ser1177 eNOS phosphorylation (1.2 0.01 fold), and total eNOS protein abundance (1.5 0.1 fold) in HUVECs from normal pregnancies. A2AAR and A2BAR activation enhanced insulin effect only on Akt (1.6 0.1 fold). LOPE only increased Ser1177 eNOS phosphorylation and total eNOS protein abundance (2.2 0.9 and 1.4 0.2 fold, respectively). Insulin blocked LOPE-increased Ser1177 eNOS phosphorylation. A2AAR and A2BAR activation did not change insulin effect on Akt and p44/42mapk, whereas A2AAR antagonist reversed insulin-decreased Ser1177 eNOS phosphorylation and increased total eNOS protein abundance in LOPE. Insulin dilation of umbilical veins from normal pregnancies was lower (14 2%, maximal relaxation (Rmax)) in the presence of A2AAR. LOPE reduced insulin dilation (18 3% Rmax), which was restored by A2AAR antagonists. Insulin increased L-citrulline content (5.3 0.3 fold), a phenomenon blocked by A2AAR and A2BAR antagonists in normal pregnancies. LOPE increased L-citrulline content, which was unaltered by insulin in absence of A2AAR and A2BAR antagonists. However, insulin increased L-citrulline content in the presence of A2AAR antagonists, but blocked by A2BAR antagonists in LOPE. Conclusion: The reduced foetoplacental vascular response to insulin in LOPE involves A2AAR activation, a phenomenon counteracted by A2BAR activation. doi:10.1016/j.preghy.2016.08.147
Clinical science 66 Renal long-term follow up after pregnancy and risk of chronic kidney disease Long term consequences for mother and child Thais Alquezar Facca, Amelia Rodrigues Pereira, Michele Tiveron Passos, Larissa Fatima Santos, Eduardo Brosco Fama, Guilherme Silva Junior, Jussara Leiko Sato, Sonia Nishida, Gianna Mastroianni Kirsztajn, Nelson Sass (UNIFESP, Sao Paulo, SP, Brazil) Introduction: Glomerular filtration rate (GFR) increases during pregnancy and some incipient nephropathies may appear after this period, especially when gestational hypertension (GH) is present.
Objectives: Renal evaluation in women over 10 years after pregnancy. Methods: Total of 30 volunteer patients whose last childbirth was 10–20 years ago. Renal parameters assessed were: urinary excretion of retinol-binding protein (RBP), urine protein/creatinine ratio (PCR), Urine albumin/creatinine ratio (ACR), serum creatinine, serum C cystatin, vitamin D (25OHD), serum uric acid, vascular endothelial growth factor (VEGF), estimated GFR (eGFR) based on Modification of Diet in Renal Disease (MDRD) Study and the Chronic Kidney Disease Epidemiology Collaboration equation – CKD-EPI Creatinine, 2009 (CKD-EPI crea), CKD-EPI Cystatin C, 2012 (CKD-EPI cys) and CKD-EPI Creatinine-Cystatin C, 2012 (CKD-EPI crea-cys) equations. Results: The average age was 45.5 years old, 56.6% were afro descendant, and mean time after last labor was 13.7 years. Approximately 6.6% had type 2 diabetes and 30.3% had chronic hypertension (CH) after pregnancy (among them 90% had GH). Mean body mass index (BMI) was 29.1 kg/m2 (overweight), although all patients with CAH were obese. Mean value of cystatin C was 1.45 mg/L (elevated in 20%), RBP 0.30 mg/L (high level in only one case), PCR 0.02 g/g, ACR 6.91 mg/g creatinine (high level in only one case), 25OHD 24.34 ng/mL (deficiency in 33.3%), serum uric acid 4.50 mg/dL (high level in only one case), serum creatinine 0.73 mg/ dL (normal level in all cases), VEGF 312.04 pg/mL (high level in two cases). Mean clearance of creatinine using MDRD was 93.5 mL/min/1.73 m2, CKD-EPI crea 101.7 mL/min/1.73 m2, CKD-EPI cys 49.0 mL/min/1.73 m2 and CKD-EPI crea-cys 68.3 mL/ min/1.73 m2 (about 30% less when compared with the equations without cystatin C, then 26.6% had eGFR < 60 mL/min/1.73 m2). None of the patients had ever been assisted by a nephrologist. Conclusions: Obesity, CH, 25OHD deficiency, high levels of serum cystatin C and low eGFR may appear years after pregnancy, especially if GH was present. Women who had GH should have long-term follow up with nephrologist to have a more complete and regular renal evaluation. The eGFR by combined creatinine-cystatin C equation seems to be better than other formulas and it could be useful to detect early chronic kidney disease. Fapesp n° 2014/00213-7 doi:10.1016/j.preghy.2016.08.148
Basic science 67 Application of a novel RUPP model to C57BL/6J mice Animal models Tomofumi Fushima, Nobuyuki Takahashi (Tohoku University Graduate School of Pharmaceutical Sciences, Sendai Japan) Introduction: Reduction of uterine blood flow is useful for studying pathogenesis of preeclampsia (PE). Although reduced uterine perfusion pressure (RUPP) is widely used as a model of PE in rats, investigating the genetics of PE has been slow because it has been difficult to make a useful RUPP model in mice. Moreover, rat model of RUPP does not allow one to measure BP using tail cuffs. We established a novel RUPP model of PE in outbred ICR mice, which allows one to evaluate BP using tail cuff (Fushima et al. PLOS ONE, 2016). Objectives: The aim of the present study is to develop RUPP model of C57BL/6J (B6) mice, a widely used strain in genetically engineered mice. Methods: We bilaterally ligated ovarian vessels distal to ovarian branches, uterine vessels, or both in B6 mice at 14.5 dpc. BP, renal phenotype and pregnancy outcome were analyzed. Results: Unlike the RUPP model in ICR mice, ligation of ovarian vessels or uterine vessels caused miscarriages in B6 mice. We next
Abstracts / Pregnancy Hypertension: An International Journal of Women’s Cardiovascular Health 6 (2016) 178–252
tied uterine vessels with a nylon thread, followed by removal of the thread to provide a small space for a little blood flow. This caused significant fetal growth restriction. Different from ICR mice BP changes were insignificant. Conclusions: Pregnancy outcome caused by RUPP is different depending on the strains of mice. This RUPP model is expected to be useful for investigating pathogenesis of PE and FGR in genetically engineered mice and for evaluating new therapies for these conditions.
211
(commonly used when anaesthesia required in women with preeclampsia), systolic BP decreased by 1.4 mmHg when animals were on labetalol (n = 3, NS), decreased by 7.2 mmHg when animals were on methyldopa (n = 2, NS) and increased by 2.8 mmHg when animals were on hydralazine (n = 3, NS) as compared to systolic BP during propofol anaesthesia prior to receiving any medication. Conclusion: These results show that in the two anaesthetic agents studied so far the greatest reduction in BP was achieved with hydralazine during ketamine anaesthesia and methyldopa during propofol anaesthesia. It is likely that antihypertensive treatment affects blood pressure during anaesthesia, and that the interaction between antihypertensive and anaesthetic agent is of clinical importance in managing sudden, unexpected and severe rises in BP in women with preeclampsia. doi:10.1016/j.preghy.2016.08.150
Basic science doi:10.1016/j.preghy.2016.08.149
69 Oxidative stress induces inflammatory response in placental explants
Clinical science
Immune and inflammatory mechanisms Priscila Rezeck Rezeck Nunes a, Mariana Romao-Veiga b, Mariana Leticia Matias a, Vanessa Rocha Ribeiro a, Jose Carlos Peracoli a, Jose Ricardo Rodrigues a, Leandro de Oliveira a (a Botucatu Medical School, Botucatu, SP, Brazil, b Institute of Biosciences, Botucatu, SP, Brazil)
68 Acute blood pressure response to antihypertensives during anaesthesia in an experimental model of preeclampsia Anesthesia – Critical care – Acute complications Suzanne Pears a, Neroli Sunderland b, Alicia Dennis c, Shirlene Lim d, Katrina Chau a, Shikha Aggarwal e, Scott Heffernan b, Ryan Downey f, Robert Ogle g, John Thompson h, Jim Iliopoulos i, Annemarie Hennessy e, Angela Makris e (a School of Medicine, University of Sydney, Sydney, Australia, b Royal Prince Alfred Hospital, Sydney, Australia, c The Royal Women’s Hospital, Melbourne, Australia, d Heart Research Institute, Sydney, Australia, e School of Medicine, Western Sydney University, Sydney, Australia, f Department of Anaesthetics, Royal Prince Alfred Hospital, Sydney, Australia, g Department of Obstetrics, Royal Prince Alfred Hospital, Sydney, Australia , h Department of Surgery, Royal Prince Alfred Hospital, Sydney, Australia, i Department of Vascular Surgery, Liverpool Hospital, Sydney, Australia) Introduction: Increased blood pressure (BP), especially sudden, unexpected and severe rises in women with preeclampsia considerably increases the risk of peripartum complications. These risks are even more acute in the context of general anaesthesia. Objectives: To assess the effects of antihypertensive drugs on short and long term blood pressure variability when used concurrently with propofol and ketamine anaesthesia. Methods: Blood Pressure was measured during anaesthesia in six, pregnant Papio hamadryas (baboons) with experimental preeclampsia (EPE). Animals were given antihypertensives commonly used to manage preeclampsia (labetalol, methyldopa and hydralazine) at equipotent doses equivalent to mild starting dose rates commonly used in women with preeclampsia. Results: When anaesthetised with ketamine (most commonly used anaesthetic agent in EPE), systolic BP increased significantly by 4.9 mmHg when animals were on labetalol (n = 3, p < 0.05), decreased by 0.5 mmHg when animals were on methyldopa (n = 3, NS) and decreased by 4.9mmHg when animals were on hydralazine (n = 2, NS) as compared to systolic BP under ketamine anaesthesia prior to receiving any medication. With propofol anaesthesia
Introduction: Oxidative stress arises from imbalance between generation of reactive oxygen species and anti-oxidant capacity of specific tissues. This alteration has been correlated with some disorders in pregnancy, mainly with pre-eclampsia (PE). In PE, placental oxidative stress contributes with the intense inflammatory response of the disease and supposed pathways related to this alteration include the formation of Inflammasome complexes that mediate the maturation of pro-inflammatory cytokines. Objectives: This study aims to evaluate if the treatment of placental explants with hydrogen peroxide (H2O2) is able to induce inflammasome complexes in placentas from normotensive pregnant women and then increase inflammatory cytokines expression. Methods: Placental explants were obtained from normotensive pregnant women undergoing elective cesarean section at 39 gestational weeks (N = 10). Placental explants were cultured in different concentrations of H2O2 (10, 100, 1000, 2000 and 10,000 lM) for 4 h. Explant controls were cultured only supplemented medium. Gene expression for cytokines (IL-1b, IL-10 and TNF-aÞ and Caspase-1 were determined by RT-qPCR. Differences between groups were analyzed by non-parametric tests with significance level set at 5%. Results: Explants cultured with 1000, 2000 and 10,000 lM of H2O2 showed lower IL-10 mRNA expression than controls. On the contrary, higher gene expression of TNF-a was observed in the same concentrations. Interestingly, IL-1b mRNA expression was higher in the presence of 10 and 100 lM H2O2 concentrations than in 10,000 lM. IL-1b mRNA expression was also higher in cultures with 100 lM H2O2 concentration than in 1000 and 2000 lM. Gene expression of Caspase-1 was increased in cultures with 100 lM of H2O2 concentration. Conclusion: These results suggest that the oxidative stress caused by H2O2 can activate caspase-1 wich participates in inflammasomes pathway and consequently increases the release of proinflammatory cytokines, such as IL-1b and TNF-a. In addition, different responses among different concentrations of H2O2 raise the
Abstracts / Pregnancy Hypertension: An International Journal of Women’s Cardiovascular Health 6 (2016) 178–252
Preeclampsia courses with endothelial and insulin resistance. Insulin dilates umbilical vein requiring A2A adenosine receptor (A2AAR) activation in normal pregnancies; however, whether A2AAR are involved in late-onset preeclampsia (LOPE)-reduced insulin dilation is unknown. LOPE increases maternal and foetal plasma adenosine and A2BAR expression in human umbilical vein endothelial cells (HUVECs). A2BAR involvement and insulin effect on the foetoplacental vascular function in LOPE is unknown. Aim: To evaluate A2AAR and A2BAR involvement on insulin effect on endothelial function in umbilical veins and HUVECs from LOPE. Methods: Protein abundance (total and phosphorylated) of p44/42mapk, protein kinase B/Akt (Akt), endothelial nitric oxide synthase (eNOS) were detected by Western blot. Assays were in the absence or presence (8 h) of 1 nM insulin, 30 nM CGS21680 (A2AAR agonist), 10 nM ZM241385 (A2AAR antagonist), 0.1 nM BAY606583 (A2BAR agonist), or 30 nM MRS1754 (A2BAR antagonist). Vascular response to insulin (0.1–1000 nM, 5 min) was measured in preconstricted umbilical vein rings (wire myography) in the absence or presence of adenosine (1 mM) and/or A2AAR and A2BAR antagonists. L-Citrulline level was measured by HPLC in the absence or presence (8 h) of NG-nitro- L-arginine methyl ester (100 lM) in HUVECs. Results: Insulin increased Akt (1.3 0.1 fold), p44/42mapk (1.2 0.1 fold), Ser1177 eNOS phosphorylation (1.2 0.01 fold), and total eNOS protein abundance (1.5 0.1 fold) in HUVECs from normal pregnancies. A2AAR and A2BAR activation enhanced insulin effect only on Akt (1.6 0.1 fold). LOPE only increased Ser1177 eNOS phosphorylation and total eNOS protein abundance (2.2 0.9 and 1.4 0.2 fold, respectively). Insulin blocked LOPE-increased Ser1177 eNOS phosphorylation. A2AAR and A2BAR activation did not change insulin effect on Akt and p44/42mapk, whereas A2AAR antagonist reversed insulin-decreased Ser1177 eNOS phosphorylation and increased total eNOS protein abundance in LOPE. Insulin dilation of umbilical veins from normal pregnancies was lower (14 2%, maximal relaxation (Rmax)) in the presence of A2AAR. LOPE reduced insulin dilation (18 3% Rmax), which was restored by A2AAR antagonists. Insulin increased L-citrulline content (5.3 0.3 fold), a phenomenon blocked by A2AAR and A2BAR antagonists in normal pregnancies. LOPE increased L-citrulline content, which was unaltered by insulin in absence of A2AAR and A2BAR antagonists. However, insulin increased L-citrulline content in the presence of A2AAR antagonists, but blocked by A2BAR antagonists in LOPE. Conclusion: The reduced foetoplacental vascular response to insulin in LOPE involves A2AAR activation, a phenomenon counteracted by A2BAR activation. doi:10.1016/j.preghy.2016.08.147
Clinical science 66 Renal long-term follow up after pregnancy and risk of chronic kidney disease Long term consequences for mother and child Thais Alquezar Facca, Amelia Rodrigues Pereira, Michele Tiveron Passos, Larissa Fatima Santos, Eduardo Brosco Fama, Guilherme Silva Junior, Jussara Leiko Sato, Sonia Nishida, Gianna Mastroianni Kirsztajn, Nelson Sass (UNIFESP, Sao Paulo, SP, Brazil) Introduction: Glomerular filtration rate (GFR) increases during pregnancy and some incipient nephropathies may appear after this period, especially when gestational hypertension (GH) is present.
Objectives: Renal evaluation in women over 10 years after pregnancy. Methods: Total of 30 volunteer patients whose last childbirth was 10–20 years ago. Renal parameters assessed were: urinary excretion of retinol-binding protein (RBP), urine protein/creatinine ratio (PCR), Urine albumin/creatinine ratio (ACR), serum creatinine, serum C cystatin, vitamin D (25OHD), serum uric acid, vascular endothelial growth factor (VEGF), estimated GFR (eGFR) based on Modification of Diet in Renal Disease (MDRD) Study and the Chronic Kidney Disease Epidemiology Collaboration equation – CKD-EPI Creatinine, 2009 (CKD-EPI crea), CKD-EPI Cystatin C, 2012 (CKD-EPI cys) and CKD-EPI Creatinine-Cystatin C, 2012 (CKD-EPI crea-cys) equations. Results: The average age was 45.5 years old, 56.6% were afro descendant, and mean time after last labor was 13.7 years. Approximately 6.6% had type 2 diabetes and 30.3% had chronic hypertension (CH) after pregnancy (among them 90% had GH). Mean body mass index (BMI) was 29.1 kg/m2 (overweight), although all patients with CAH were obese. Mean value of cystatin C was 1.45 mg/L (elevated in 20%), RBP 0.30 mg/L (high level in only one case), PCR 0.02 g/g, ACR 6.91 mg/g creatinine (high level in only one case), 25OHD 24.34 ng/mL (deficiency in 33.3%), serum uric acid 4.50 mg/dL (high level in only one case), serum creatinine 0.73 mg/ dL (normal level in all cases), VEGF 312.04 pg/mL (high level in two cases). Mean clearance of creatinine using MDRD was 93.5 mL/min/1.73 m2, CKD-EPI crea 101.7 mL/min/1.73 m2, CKD-EPI cys 49.0 mL/min/1.73 m2 and CKD-EPI crea-cys 68.3 mL/ min/1.73 m2 (about 30% less when compared with the equations without cystatin C, then 26.6% had eGFR < 60 mL/min/1.73 m2). None of the patients had ever been assisted by a nephrologist. Conclusions: Obesity, CH, 25OHD deficiency, high levels of serum cystatin C and low eGFR may appear years after pregnancy, especially if GH was present. Women who had GH should have long-term follow up with nephrologist to have a more complete and regular renal evaluation. The eGFR by combined creatinine-cystatin C equation seems to be better than other formulas and it could be useful to detect early chronic kidney disease. Fapesp n° 2014/00213-7 doi:10.1016/j.preghy.2016.08.148
Basic science 67 Application of a novel RUPP model to C57BL/6J mice Animal models Tomofumi Fushima, Nobuyuki Takahashi (Tohoku University Graduate School of Pharmaceutical Sciences, Sendai Japan) Introduction: Reduction of uterine blood flow is useful for studying pathogenesis of preeclampsia (PE). Although reduced uterine perfusion pressure (RUPP) is widely used as a model of PE in rats, investigating the genetics of PE has been slow because it has been difficult to make a useful RUPP model in mice. Moreover, rat model of RUPP does not allow one to measure BP using tail cuffs. We established a novel RUPP model of PE in outbred ICR mice, which allows one to evaluate BP using tail cuff (Fushima et al. PLOS ONE, 2016). Objectives: The aim of the present study is to develop RUPP model of C57BL/6J (B6) mice, a widely used strain in genetically engineered mice. Methods: We bilaterally ligated ovarian vessels distal to ovarian branches, uterine vessels, or both in B6 mice at 14.5 dpc. BP, renal phenotype and pregnancy outcome were analyzed. Results: Unlike the RUPP model in ICR mice, ligation of ovarian vessels or uterine vessels caused miscarriages in B6 mice. We next
Abstracts / Pregnancy Hypertension: An International Journal of Women’s Cardiovascular Health 6 (2016) 178–252
tied uterine vessels with a nylon thread, followed by removal of the thread to provide a small space for a little blood flow. This caused significant fetal growth restriction. Different from ICR mice BP changes were insignificant. Conclusions: Pregnancy outcome caused by RUPP is different depending on the strains of mice. This RUPP model is expected to be useful for investigating pathogenesis of PE and FGR in genetically engineered mice and for evaluating new therapies for these conditions.
211
(commonly used when anaesthesia required in women with preeclampsia), systolic BP decreased by 1.4 mmHg when animals were on labetalol (n = 3, NS), decreased by 7.2 mmHg when animals were on methyldopa (n = 2, NS) and increased by 2.8 mmHg when animals were on hydralazine (n = 3, NS) as compared to systolic BP during propofol anaesthesia prior to receiving any medication. Conclusion: These results show that in the two anaesthetic agents studied so far the greatest reduction in BP was achieved with hydralazine during ketamine anaesthesia and methyldopa during propofol anaesthesia. It is likely that antihypertensive treatment affects blood pressure during anaesthesia, and that the interaction between antihypertensive and anaesthetic agent is of clinical importance in managing sudden, unexpected and severe rises in BP in women with preeclampsia. doi:10.1016/j.preghy.2016.08.150
Basic science doi:10.1016/j.preghy.2016.08.149
69 Oxidative stress induces inflammatory response in placental explants
Clinical science
Immune and inflammatory mechanisms Priscila Rezeck Rezeck Nunes a, Mariana Romao-Veiga b, Mariana Leticia Matias a, Vanessa Rocha Ribeiro a, Jose Carlos Peracoli a, Jose Ricardo Rodrigues a, Leandro de Oliveira a (a Botucatu Medical School, Botucatu, SP, Brazil, b Institute of Biosciences, Botucatu, SP, Brazil)
68 Acute blood pressure response to antihypertensives during anaesthesia in an experimental model of preeclampsia Anesthesia – Critical care – Acute complications Suzanne Pears a, Neroli Sunderland b, Alicia Dennis c, Shirlene Lim d, Katrina Chau a, Shikha Aggarwal e, Scott Heffernan b, Ryan Downey f, Robert Ogle g, John Thompson h, Jim Iliopoulos i, Annemarie Hennessy e, Angela Makris e (a School of Medicine, University of Sydney, Sydney, Australia, b Royal Prince Alfred Hospital, Sydney, Australia, c The Royal Women’s Hospital, Melbourne, Australia, d Heart Research Institute, Sydney, Australia, e School of Medicine, Western Sydney University, Sydney, Australia, f Department of Anaesthetics, Royal Prince Alfred Hospital, Sydney, Australia, g Department of Obstetrics, Royal Prince Alfred Hospital, Sydney, Australia , h Department of Surgery, Royal Prince Alfred Hospital, Sydney, Australia, i Department of Vascular Surgery, Liverpool Hospital, Sydney, Australia) Introduction: Increased blood pressure (BP), especially sudden, unexpected and severe rises in women with preeclampsia considerably increases the risk of peripartum complications. These risks are even more acute in the context of general anaesthesia. Objectives: To assess the effects of antihypertensive drugs on short and long term blood pressure variability when used concurrently with propofol and ketamine anaesthesia. Methods: Blood Pressure was measured during anaesthesia in six, pregnant Papio hamadryas (baboons) with experimental preeclampsia (EPE). Animals were given antihypertensives commonly used to manage preeclampsia (labetalol, methyldopa and hydralazine) at equipotent doses equivalent to mild starting dose rates commonly used in women with preeclampsia. Results: When anaesthetised with ketamine (most commonly used anaesthetic agent in EPE), systolic BP increased significantly by 4.9 mmHg when animals were on labetalol (n = 3, p < 0.05), decreased by 0.5 mmHg when animals were on methyldopa (n = 3, NS) and decreased by 4.9mmHg when animals were on hydralazine (n = 2, NS) as compared to systolic BP under ketamine anaesthesia prior to receiving any medication. With propofol anaesthesia
Introduction: Oxidative stress arises from imbalance between generation of reactive oxygen species and anti-oxidant capacity of specific tissues. This alteration has been correlated with some disorders in pregnancy, mainly with pre-eclampsia (PE). In PE, placental oxidative stress contributes with the intense inflammatory response of the disease and supposed pathways related to this alteration include the formation of Inflammasome complexes that mediate the maturation of pro-inflammatory cytokines. Objectives: This study aims to evaluate if the treatment of placental explants with hydrogen peroxide (H2O2) is able to induce inflammasome complexes in placentas from normotensive pregnant women and then increase inflammatory cytokines expression. Methods: Placental explants were obtained from normotensive pregnant women undergoing elective cesarean section at 39 gestational weeks (N = 10). Placental explants were cultured in different concentrations of H2O2 (10, 100, 1000, 2000 and 10,000 lM) for 4 h. Explant controls were cultured only supplemented medium. Gene expression for cytokines (IL-1b, IL-10 and TNF-aÞ and Caspase-1 were determined by RT-qPCR. Differences between groups were analyzed by non-parametric tests with significance level set at 5%. Results: Explants cultured with 1000, 2000 and 10,000 lM of H2O2 showed lower IL-10 mRNA expression than controls. On the contrary, higher gene expression of TNF-a was observed in the same concentrations. Interestingly, IL-1b mRNA expression was higher in the presence of 10 and 100 lM H2O2 concentrations than in 10,000 lM. IL-1b mRNA expression was also higher in cultures with 100 lM H2O2 concentration than in 1000 and 2000 lM. Gene expression of Caspase-1 was increased in cultures with 100 lM of H2O2 concentration. Conclusion: These results suggest that the oxidative stress caused by H2O2 can activate caspase-1 wich participates in inflammasomes pathway and consequently increases the release of proinflammatory cytokines, such as IL-1b and TNF-a. In addition, different responses among different concentrations of H2O2 raise the