Effects of intermittent high glucose on BeWo choriocarcinoma cells in culture.

AIM: The aim of this study was to investigate the cellular effects of intermittent high glucose on the human BeWo placental choriocarcinoma cell line, used as a model of the effects of glucose fluctuation in diabetic pregnancies. MATERIALS AND METHODS: BeWo cells were subjected to three different glucose conditions for 48 h: 7 mmol/L (control), 42 mmol/L (high glucose), or 7 and 42 mmol/L glucose (intermittent, alternated every 6 h). Cell viability was assessed using cell counts, a cell proliferation assay, and a cell viability assay. Apoptosis was also studied using a terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling assay and by immunocytochemistry of fractin, the N-terminal fragment of actin, which can distinguish apoptotic from necrotic cells. Furthermore, the expression of the well-known survival factors of trophoblast cells, heparin-binding epidermal growth factor-like growth factor and leptin, was evaluated by real-time polymerase chain reaction and Western blot analyses. RESULTS: Intermittent high-glucose conditions significantly decreased cell viability and enhanced apoptosis compared with control or continuous high-glucose conditions. Furthermore, the expression of heparin-binding epidermal growth factor-like growth factor, but not that of leptin, was significantly increased under intermittent high-glucose conditions compared to its expression under either control or continuous high-glucose conditions. CONCLUSIONS: These data indicate that intermittent high glucose is more deleterious to BeWo cells than continuous high-glucose conditions. Although further in vitro and in vivo study is necessary, excess fluctuation of glucose levels in the placental circulation might be involved in the impairment of placental development leading to the placental dysfunction.

Expression of antiangiogenic prolactin fragments in the placentas of women with pregnancy induced hypertension.

It has been reported that prolactin (PRL) is cleaved to 14 or 16 kDa fragments by cathepsin D in vitro and in vivo, and that such fragments exhibit antiangiogenic and proapoptotic properties. The aim of this study was to investigate the relationship between pregnancy induced hypertension (PIH) and the placental expression of antiangiogenic PRL fragments and cathepsin D. Placental expression of PRL fragments and cathepsin D was evaluated by Western blot analysis in a group of 9 pregnant women consisting of 5 normal pregnancies and 4 severe PIH cases. Antiangiogenic PRL fragments were detected in 4 placental samples from all PIH cases but not detected in those from normal pregnancies (p<0.05). The expression of cathepsin D in PIH placentas was significantly lower than that in those without PIH (p<0.05), while the placental expression of procathepsin D was significantly greater in PIH cases than in the normal pregnancies (p<0.05). These data suggest that antiangiogenic PRL fragments in the placenta may be present only in PIH cases, and that PRL fragments in the placenta might be implicated in the pathophysiology of PIH.

Circulating levels of ciliary neurotrophic factor in normal pregnancy and preeclampsia.

Ciliary neurotrophic factor (CNTF) has been shown to decrease food intake in mouse models of obesity and to improve insulin sensitivity. It is well known that tight regulation of glucose metabolism is essential for successful gestational outcomes (e.g. fetal growth), and that abnormal insulin resistance is associated with preeclampsia (PE). To investigate the possibility that CNTF might be involved in the regulation of insulin resistance during pregnancy, circulating levels of CNTF were assessed in non-pregnant, normal pregnant, postpartum, and pregnant women with PE. Sera from healthy non-pregnant women (n = 10), pregnant women (n = 30:1st trimester; n = 10, 2nd trimester n = 10; 3rd trimester; n = 10), postpartum women (n = 10), and patients with PE (n = 11) were studied with Western blotting. Circulating CNTF was detected by Western blotting, and the levels of CNTF in pregnant women were decreased as compared with those in non-pregnant women, and tended to decrease as pregnancy progressed. A significant decrease was found in PE as compared with normal pregnancy. Circulating CNTF might be associated with physiological and abnormal insulin resistance during pregnancy.

Successful management of anti-Jra alloimmunization in pregnancy: a case report.

BACKGROUND: Hemolytic disease of the fetus/newborn due to Jr(a) immunization is very rare and considered to be mild, and only routine obstetrical care is recommended for pregnant women sensitized to the Jr(a) antigen. CASE REPORT: A 20-year-old nulliparous woman was referred to our hospital for perinatal management. Her indirect Coombs test was positive for anti-Jr(a) antibody (1:64). At 33 weeks' gestational age, we observed that fetal growth was mildly restricted and the peak systolic velocity of the fetal middle cerebral artery (PSV-MCA) was above the upper limit of the reference range (1.55 multiples of the median). Amniocentesis was also carried out and the DeltaOD450 value was in the lower mid-zone of the Liley curve. We continued to carefully observe the patient because we observed PSV-MCA values within 1.50-1.60 multiples of the median and no other findings of fetal anemia. She vaginally delivered a female infant weighing 2,136 g at 37 weeks' gestational age. The infant received treatment with both iron and recombinant erythropoietin without developing hyperbilirubinemia and blood transfusion. CONCLUSION: PSV-MCA should be monitored for the detection of fetal anemia, even in pregnant women sensitized to some antigens for which only routine obstetrical care is recommended.

Elevated level of serum retinol-binding protein 4 in pregnancy-induced hypertension.

AIM: Recent progress in adiposcience has revealed several important adipose-tissue-originated factors, so-called adipokines. Retinol-binding protein 4 (RBP4), a protein expressed and secreted by adipocytes, has been identified as a novel regulator of insulin resistance. Physiological insulin resistance occurs during the pregnancy of mammals to accommodate fetal growth, and it has been suggested that insulin resistance and hyperinsulinemia might also be associated with pregnancy-induced hypertension (PIH). In order to shed light on the role of RBP4 during pregnancy, we attempted to assess RBP4 levels during pregnancy. Fetal growth could be affected by aberrant regulation of RBP4 levels in fetal circulation per se, so we examined the RBP4 levels in cord blood samples of growth restricted cases. METHODS: Circulating RBP4 levels were examined in non-pregnant and pregnant healthy women using Western blotting. Also, RBP4 levels in normal pregnancy and PIH were quantitated using Dot-blot analysis. RBP4 levels in cord blood samples also were evaluated in selected cases. RESULTS: RBP4 levels tended to decrease after early gestation with no obvious difference between mid- to late-gestation. RBP4 levels were increased in pregnant women with PIH compared with normal pregnancies (P < 0.01). RBP4 levels were decreased in the cord blood of PIH pregnancies and light-for-dates infants without obvious causes for intrauterine growth restriction compared with normal pregnancies (P < 0.01). CONCLUSIONS: These results suggest that circulating RBP4 could be elevated in PIH, where maternal glucose metabolism is perturbed, and that RBP4 levels in cord blood might be closely associated with fetal growth.

Relation between neonatal jaundice and oncostatin M, hepatocyte growth factor and soluble gp130 levels in umbilical cord.

OBJECTIVES: Oncostatin M (OSM), hepatocyte growth factor (HGF) and soluble glycoprotein 130 (sgp130) have been demonstrated to be involved in fetal liver development. In this study, we examined the relation between neonatal jaundice and OSM, gp130 or HGF levels in umbilical cord blood. DESIGN: A cross-sectional study of 160 neonates born at Okayama University Hospital. SETTING: Tertiary referral center serving a population of three million. METHODS: The serum concentrations of OSM, HGF and sgp130 in umbilical venous cord blood were measured and compared with the clinical records of these neonates. RESULTS: The HGF level was correlated to both gestational week (p=0.007) and birthweight (p=0.015), as well as to total bilirubin on Day 1 (p=0.032). In preterm neonates, the HGF levels were lower in neonates who received phototherapy (photo group) than in those who did not receive phototherapy (non-photo group) (p=0.002). In contrast, in term neonates, OSM levels were lower in the photo group than in the non-photo group (p=0.0003). CONCLUSIONS: These findings suggest that the phototherapy requirement for neonatal jaundice may be associated with lower umbilical HGF concentration in preterm neonates and with lower umbilical OSM concentration in term neonates.

Hypoadiponectinemia and circulating angiogenic factors in overweight patients complicated with pre-eclampsia.

OBJECTIVE: Obesity is a risk factor for pre-eclampsia, and adiponectin is an important adipocyte-derived hormone that might protect the endothelium. Recent reports have underlined importance of circulating angiogenic factors for pathophysiology of pre-eclampsia. Here we examined whether adiponectin in conjunction with angiogenic factors plays some roles in the pathophysiology of pre-eclampsia. STUDY DESIGN: We measured serum concentrations of adiponectin and angiogenic factors, vascular endothelial growth factor, placental growth factor, and the soluble vascular endothelial growth factor receptors, soluble fms-like tyrosine Kinase 1 (sFlt-1) and soluble fetal liver kinase 1 (sFlk-1), in women with pre-eclampsia and healthy pregnant women. We also investigated the differences of these factors in overweight and normal-weight patients with pre-eclampsia. RESULTS: Significant correlations were found among the circulating adiponectin and angiogenic factors placental growth factor (R = 0.772, P = .0012) and sFlt-1 (R = 0.787, P = .0005); while, no correlation was found between adiponectin and sFlk-1 (R = 0.3, P = .3434) in patients with pre-eclampsia. Overweight patients with pre-eclampsia showed significantly lower adiponectin levels and a mild imbalance of circulating angiogenic factors, compared with normal-weight patients with pre-eclampsia. CONCLUSION: Overweight women with pre-eclampsia appeared to have lower levels of adiponectin and sFlt1 and higher levels of placental growth factor than women with pre-eclampsia who were of normal weight. These results suggest that hypoadiponectinemia might be involved in the pathophysiology of overweight patients with pre-eclampsia.

Circulating angiogenic factors in preeclampsia, gestational proteinuria, and preeclampsia superimposed on chronic glomerulonephritis.

OBJECTIVE: Recently, reports have indicated that the imbalance of circulating angiogenic factors is important in the onset of preeclampsia. In this study we investigated angiogenic factor levels in women with preeclampsia, preeclampsia superimposed on chronic glomerulonephritis, gestational proteinuria, and normal pregnancies. STUDY DESIGN: We measured several circulating angiogenic factors, placental growth factor, vascular endothelial growth factor, soluble fms-like tyrosine kinase-1, and soluble fetal liver kinase-1. RESULTS: Compared with women with normal pregnancies, placental growth factor concentrations were lower, and soluble fms-like tyrosine kinase 1 concentrations were higher in women with preeclampsia and gestational proteinuria; soluble fms-like tyrosine kinase 1 concentrations were also higher in women with preeclampsia superimposed on chronic glomerulonephritis. CONCLUSION: There were obvious differences in the levels of circulating angiogenic factors placental growth factor and soluble fms-like tyrosine kinase 1 among preeclampsia, gestational proteinuria, and controls. The imbalance of angiogenic factors is important in the onset of preeclampsia superimposed on chronic glomerulonephritis.