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761: Low oxygen tension upregulates expression of HTRA1 protein in the RCHO-1 trophoblast cell line
Poster Session V
Fetus Diabetes, etc
761 Low oxygen tension upregulates expression of HTRA1 protein in the RCHO-1 trophoblast cell line Funmi Ajayi1, Douglas Kniss1 1
The Ohio State University, Dept of Obstetrics and Gynecology, Div of Maternal Fetal Medicine, Columbus, Ohio
OBJECTIVE: Recent work has demonstrated that increased levels of High temperature requirement factor A1 (HtrA1) inhibits migration and invasion of trophoblast cells. Preeclampsia is thought to be a result of hypoxia due to shallow trophoblast invasion. We seek to determine if HtrA1 protein expression is up-regulated in a hypoxic environment. STUDY DESIGN: The rat trophoblast cell line (RCHO-1) was cultured in varying concentrations of oxygen: typical tissue culture oxygen tension of 21%, 2% to simulate hypoxia, and 6% to simulate physiologic in utero oxygen tension. At several time points during the differentiation process, cells were harvested and HtrA1 protein expression was evaluated by immunoblotting. RESULTS: We found that 6% oxygen tension resulted in down-regulation of HtrA1 protein expression. By day 7, HtrA1 protein expression was increased in RCHO-1 cells cultured in 2% oxygen in comparison to 21% oxygen. CONCLUSION: These results indicate that there is minimal HtrA1 protein expression in RCHO-1 trophoblast cells cultured in physiologic in utero oxygen tension. As elevated levels of HtrA1 have been noted in placentas of women with early onset preeclampsia, it is possible that the hypoxic intrauterine microenvironment in preeclamptic pregnancies contributes to increased levels of HtrA1 which then results in further inhibition of trophoblast cell migration and invasion.
www.AJOG.org STUDY DESIGN: We conducted a retrospective study from adminis-
trative data obtained from the linked California discharge database and vital statistics data sets between 1991 and 2001. Singleton pregnancies with pregestational diabetes were identified and subcategorized into groups with hypertensive diseases of pregnancy (PIH), chronic hypertension (CHTN), or neither. Data was abstracted for obstetric and neonatal outcomes. Multivariate analysis was performed using the chi-square test and adjusted for confounding factors. RESULTS: We identified 11,688 diabetic subjects of a total of 2,350,085 deliveries occurring during the specified time period. Compared to diabetes alone, patients with both diabetes and PIH had an increased risk for maternal cardiorespiratory disorders (p⫽.005), renal failure (p⫽.004), and delivery at GA ⬍34 wks (p⬍.001). For patients with diabetes and CHTN, there was an increased risk for cardiorespiratory disorders (p⫽.002), renal failure (p⫽.007), placental abruption (p⫽0.003), delivery at GA ⬍34 wks (p⬍.001), and neonatal death (p⫽ 0.018). Between the three groups, we found no increased risk for in-hospital maternal death, CNS events, pulmonary embolus, or amniotic fluid embolism. We found an incremental risk between normotensives, those with PIH, and those with CHTN in fetal outcomes such as SGA (p⬍.001 and p⬍.001 respectively), IUGR (p⬍.00, p⬍.001), LBW ⬍2500g (p⬍.00, p⬍.001), VLBW ⬍1500g (p⫽.001, p⬍.001). CONCLUSION: Among women with preexisting diabetes who have a comorbid hypertensive disorder during pregnancy, both maternal and fetal outcomes are adversely affected. We hypothesize that these observed outcomes are related to the level of underlying vasculopathy. This information could be useful for pre-pregnancy counselling. 0002-9378/$ – see front matter • doi:10.1016/j.ajog.2009.10.779
763 Levels of oxidized proteins are higher in first trimester human placenta from women at high risk for recurrent preeclampsia Mark Alanis1, Danyelle Townsend2, Elizabeth Steadman1, Yefim Manevich3, Laura Goetzl1 1 Medical University of South Carolina, Obstetrics and Gynecology, Charleston, South Carolina, 2Medical University of South Carolina, Pharmaceutical and Biomedical Sciences, Charleston, South Carolina, 3 Medical University of South Carolina, Cell and Molecular Pharmacology and Experimental Therapeutics, Charleston, South Carolina
OBJECTIVE: To compare the level of villous oxidative stress between
Oxygen Tension and HtrA1 Protein Expression 0002-9378/$ – see front matter • doi:10.1016/j.ajog.2009.10.778
762 Preexisting diabetes with concurrent hypertensive disorders in pregnancy is associated with worse pregnancy outcomes Bonnie Cheung1, Guibo Xing2, Dena Towner1 1 University of California, Davis, Sacramento, California, 2University of California, Davis, Obstetrics and Gynecology, Sacramento, California
OBJECTIVE: Diabetes and hypertensive disorders in pregnancy are associated with significant morbidity. Our goal was to analyze for any increased risk in rare morbidities in diabetics who also have a hypertensive disorder.
S274
women at high risk and low risk of recurrent preeclampsia by measuring protein sulfhydryls in first trimester human placenta homogenates. STUDY DESIGN: Multiparous women at high (n⫽6) and low (n⫽6) risk for preeclampsia were characterized by a history of severe preeclampsia (recurrence risk 25-65%) or only normal, term pregnancies (1% risk), respectively. Controls were matched by race, gestational age, and smoking history. First trimester placentas were collected at the time of elective pregnancy terminations. The redox status was assessed using the molecular probe, ThioGlo-1 (Calbioch). The emission (513 nm, excitation at 379 nm) of each sample was recorded for 1 min (background) before and 2 min after an addition of 5 M (final concentration) of ThioGlo-1. Each sample fluorescence saturation value corresponds to a concentration of free sulfhydryls, and three independent measurements per condition were averaged. Group comparisons were made using parametric and non-parametric bivariable analyses. RESULTS: There were no differences in age, ethnicity, BMI, or parity between high risk women and matched controls (p ⬎ .05). The level of ThioGlo-1 emission per mcg of protein was lower among women at high risk for recurrent preeclampsia compared to women at low risk (Figure, p ⫽ .03).
American Journal of Obstetrics & Gynecology Supplement to DECEMBER 2009
Fetus Diabetes, etc
761 Low oxygen tension upregulates expression of HTRA1 protein in the RCHO-1 trophoblast cell line Funmi Ajayi1, Douglas Kniss1 1
The Ohio State University, Dept of Obstetrics and Gynecology, Div of Maternal Fetal Medicine, Columbus, Ohio
OBJECTIVE: Recent work has demonstrated that increased levels of High temperature requirement factor A1 (HtrA1) inhibits migration and invasion of trophoblast cells. Preeclampsia is thought to be a result of hypoxia due to shallow trophoblast invasion. We seek to determine if HtrA1 protein expression is up-regulated in a hypoxic environment. STUDY DESIGN: The rat trophoblast cell line (RCHO-1) was cultured in varying concentrations of oxygen: typical tissue culture oxygen tension of 21%, 2% to simulate hypoxia, and 6% to simulate physiologic in utero oxygen tension. At several time points during the differentiation process, cells were harvested and HtrA1 protein expression was evaluated by immunoblotting. RESULTS: We found that 6% oxygen tension resulted in down-regulation of HtrA1 protein expression. By day 7, HtrA1 protein expression was increased in RCHO-1 cells cultured in 2% oxygen in comparison to 21% oxygen. CONCLUSION: These results indicate that there is minimal HtrA1 protein expression in RCHO-1 trophoblast cells cultured in physiologic in utero oxygen tension. As elevated levels of HtrA1 have been noted in placentas of women with early onset preeclampsia, it is possible that the hypoxic intrauterine microenvironment in preeclamptic pregnancies contributes to increased levels of HtrA1 which then results in further inhibition of trophoblast cell migration and invasion.
www.AJOG.org STUDY DESIGN: We conducted a retrospective study from adminis-
trative data obtained from the linked California discharge database and vital statistics data sets between 1991 and 2001. Singleton pregnancies with pregestational diabetes were identified and subcategorized into groups with hypertensive diseases of pregnancy (PIH), chronic hypertension (CHTN), or neither. Data was abstracted for obstetric and neonatal outcomes. Multivariate analysis was performed using the chi-square test and adjusted for confounding factors. RESULTS: We identified 11,688 diabetic subjects of a total of 2,350,085 deliveries occurring during the specified time period. Compared to diabetes alone, patients with both diabetes and PIH had an increased risk for maternal cardiorespiratory disorders (p⫽.005), renal failure (p⫽.004), and delivery at GA ⬍34 wks (p⬍.001). For patients with diabetes and CHTN, there was an increased risk for cardiorespiratory disorders (p⫽.002), renal failure (p⫽.007), placental abruption (p⫽0.003), delivery at GA ⬍34 wks (p⬍.001), and neonatal death (p⫽ 0.018). Between the three groups, we found no increased risk for in-hospital maternal death, CNS events, pulmonary embolus, or amniotic fluid embolism. We found an incremental risk between normotensives, those with PIH, and those with CHTN in fetal outcomes such as SGA (p⬍.001 and p⬍.001 respectively), IUGR (p⬍.00, p⬍.001), LBW ⬍2500g (p⬍.00, p⬍.001), VLBW ⬍1500g (p⫽.001, p⬍.001). CONCLUSION: Among women with preexisting diabetes who have a comorbid hypertensive disorder during pregnancy, both maternal and fetal outcomes are adversely affected. We hypothesize that these observed outcomes are related to the level of underlying vasculopathy. This information could be useful for pre-pregnancy counselling. 0002-9378/$ – see front matter • doi:10.1016/j.ajog.2009.10.779
763 Levels of oxidized proteins are higher in first trimester human placenta from women at high risk for recurrent preeclampsia Mark Alanis1, Danyelle Townsend2, Elizabeth Steadman1, Yefim Manevich3, Laura Goetzl1 1 Medical University of South Carolina, Obstetrics and Gynecology, Charleston, South Carolina, 2Medical University of South Carolina, Pharmaceutical and Biomedical Sciences, Charleston, South Carolina, 3 Medical University of South Carolina, Cell and Molecular Pharmacology and Experimental Therapeutics, Charleston, South Carolina
OBJECTIVE: To compare the level of villous oxidative stress between
Oxygen Tension and HtrA1 Protein Expression 0002-9378/$ – see front matter • doi:10.1016/j.ajog.2009.10.778
762 Preexisting diabetes with concurrent hypertensive disorders in pregnancy is associated with worse pregnancy outcomes Bonnie Cheung1, Guibo Xing2, Dena Towner1 1 University of California, Davis, Sacramento, California, 2University of California, Davis, Obstetrics and Gynecology, Sacramento, California
OBJECTIVE: Diabetes and hypertensive disorders in pregnancy are associated with significant morbidity. Our goal was to analyze for any increased risk in rare morbidities in diabetics who also have a hypertensive disorder.
S274
women at high risk and low risk of recurrent preeclampsia by measuring protein sulfhydryls in first trimester human placenta homogenates. STUDY DESIGN: Multiparous women at high (n⫽6) and low (n⫽6) risk for preeclampsia were characterized by a history of severe preeclampsia (recurrence risk 25-65%) or only normal, term pregnancies (1% risk), respectively. Controls were matched by race, gestational age, and smoking history. First trimester placentas were collected at the time of elective pregnancy terminations. The redox status was assessed using the molecular probe, ThioGlo-1 (Calbioch). The emission (513 nm, excitation at 379 nm) of each sample was recorded for 1 min (background) before and 2 min after an addition of 5 M (final concentration) of ThioGlo-1. Each sample fluorescence saturation value corresponds to a concentration of free sulfhydryls, and three independent measurements per condition were averaged. Group comparisons were made using parametric and non-parametric bivariable analyses. RESULTS: There were no differences in age, ethnicity, BMI, or parity between high risk women and matched controls (p ⬎ .05). The level of ThioGlo-1 emission per mcg of protein was lower among women at high risk for recurrent preeclampsia compared to women at low risk (Figure, p ⫽ .03).
American Journal of Obstetrics & Gynecology Supplement to DECEMBER 2009