[show abstract][hide abstract] ABSTRACT: Offspring of pregnancy complicated with preeclampsia are at high risk for hypertension, stroke and possibly obesity. The mechanisms behind the association of intrauterine exposure to preeclampsia and high risk of health problems in the later life remain largely unknown. The aims of the current investigation were to determine the changes in DNA methylation at IGF2 and GNAS DMR in offspring of preeclamptic pregnancy and to explore the possible mechanisms underlying the association between maternal preeclampsia and high risk for health problems in the later life of their offspring.
Umbilical cord blood was taken from infants born to women of preeclampsia (n=56), gestational hypertension (n=23) and normal pregnancy (n=81). DNA methylation levels of IGF2 and GNAS DMR were determined by Massarray quantitative methylation analysis. Methylation levels at IGF2 DMR were significantly lower in preeclampsia than normal pregnancy. The average methylation level at IGF2 DMR was significantly correlated with preeclampsia even after birth weight, maternal age, gestational age at delivery and fetal gender were adjusted. The difference in methylation level was not significantly different between mild and severe preeclampsia. The methylation level at GNAS DMR was not significantly correlated with birth weight, maternal age, gestational age at delivery, fetal gender, preeclampsia or gestational hypertension.
We concluded preeclampsia induced a decrease in methylation level at IGF 2 DMR, and this might be among the mechanisms behind the association between intrauterine exposure to preeclampsia and high risk for metabolic diseases in the later life of the infants.
[show abstract][hide abstract] ABSTRACT: Preadipocyte factor-1(Pref-1), an inhibitor of adipocyte differentiation, is increased in fetal blood of small for gestational age (SGA) and is considered a factor involved in determining adiposity and associated with high risk of metabolic diseases in adulthood. Preeclampsia is a condition closely associated with SGA, however, the alteration of Pref-1 of in fetuses of preeclampsia remains unknown. The aims of the current investigation were to clarify the alteration of serum Pref-1 in fetuses of preelamptic pregnancy and to explore possible role of Pref-1 in metabolic diseases in late life.
Cord blood samples were taken at birth from 45 fetuses of normal pregnancy, 16 of gestational hypertension, 29 of mild preeclampsia and 29 of severe preeclampsia. Serum Pref-1 concentrations were measured with ELISA.
There were significant differences in cord blood Pref-1 and neonatal birth weight among normal pregnancy, gestational hypertension, mild and severe preeclampsia (F=8.557, P<0.001 for pref-1; F=38.405, P<0.001 for birth weight). Serum pref-1 was significantly higher while birth weight were lower in severe preeclampsia than normal pregnancy, gestational hypertension and mild preeclampsia respectively (P<=0.001 for all). However, either serum pref-1 or birth weight did not significantly differ among normal pregnancy, gestational hypertension and mild preeclampsia (P>0.05 for all). Fetal pref-1 concentration was significantly negatively correlated with birth weight (R(2)=0.175, P=0.027 for severe preeclampsia; R(2)=0.209, P<0.001 for preeclampsia; R(2)=0.25, P<0.001 for all subjects).
Increased serum pref-1 was demonstrated in fetuses of preeclampsia- complicated pregnancy, and it may be proposed that pref-1 is among the possible mediators leading to high risk of metabolic diseases in adulthood.
Clinica chimica acta; international journal of clinical chemistry 06/2013; · 2.54 Impact Factor
[show abstract][hide abstract] ABSTRACT: Intrauterine transmission of hepatitis B virus (HBV) is the main cause of the high prevalence of HBV in endemic areas; however, the mechanisms underlying intrauterine transmission of HBV remain unknown. To explore the role of mannose-binding lectin (MBL), a pattern recognition molecule of the innate immune system, in intrauterine transmission of HBV, we determined MBL levels using an enzyme-linked immunosorbent assay (ELISA) in cord serum of 7 intrauterine-infected neonates and 30 non-infected neonates born to HBV-positive mothers, and 30 control neonates born to HBV-negative mothers. We observed significant differences in cord serum MBL levels among the three groups (P < 0.001). Non-infected neonates had significantly higher MBL levels than controls (P < 0.001), and intrauterine-infected neonates had significantly lower serum MBL levels than non-infected neonates (P < 0.001). However, serum MBL levels were not significantly different between intrauterine-infected neonates and controls (P = 0.800). Our results indicate that maternal HBV infection induces an increase in fetal MBL levels and the absence of this increase is possibly associated with intrauterine transmission of HBV, suggesting that MBL plays a role in intrauterine transmission of HBV.
Japanese journal of infectious diseases. 01/2013; 66(5):391-3.
[show abstract][hide abstract] ABSTRACT: To clarify the alterations of myostatin, a member of the transforming growth factor-β superfamily, and follistatin-like 3 (FSTL3), a binding protein for myostatin, in pre-eclamptic women.
Samples of blood and placenta were collected from 40 pre-eclamptic women and 40 controls. The serum level and placental expression of FSTL3 and myostatin were determined with enzyme-linked immunosorbent assay, real-time polymerized chain reaction and western blotting.
The serum levels of myostatin and FSTL3 were significantly higher in pre-eclamptic women than in the controls (P < 0.001 for both). Placental expression of myostatin and FSTL3 were also significantly increased in the pre-eclamptic placenta compared with that of the controls (P < 0.001 for both); however, there were no significant differences in myostatin or FSTL3 in either the maternal serum or the placenta in women with mild or severe pre-eclampsia (P > 0.05 for both).
The serum levels and placental expression of myostatin and FSTL3 are elevated in pre-eclampsia, suggesting the role of myostatin and its binding protein in pre-eclampsia.
Journal of Obstetrics and Gynaecology Research 05/2012; 38(7):988-96. · 0.84 Impact Factor
[show abstract][hide abstract] ABSTRACT: To investigate whether bile acids transporters organic anion transporting polypeptides 1A2 (OATP1A2), organic anion transporting polypeptides 1B1 (OATP1B1), organic anion transporting polypeptides 1B3 (OATP1B3) were differently expressed in placenta of intrahepatic cholestasis of pregnancy (ICP).
Thirty pregnant women with ICP were recruited and 30 normal pregnant women served as control. The expression of mRNA and protein were analyzed by real-time PCR and Western blotting. The localization of OATP1A2, OATP1B1, OATP1B3 were investigated by immunohistochemistry.
The expression of mRNA and protein of both OATP1A2 and OATP1B3 were significantly lower in ICP placenta than normal placenta (P < 0.05). OATP1B1 mRNA was detected by RT-PCR in 8 ICP placentas and 7 control placentas, but protein expression of OATP1B1 was not found in any of the 60 placentas. By immunohistochemistry we found that OATP1A2 was obviously localized to vasculo-syncytial membrane (VSM) and apical surface of syncytiotrophoblasts, while OATP1B3 was localized to VSM of the syncytiotrophoblasts.
The expression of OATP1A2 and OATP1B3 in placenta decreased in ICP. The down-regulation of these transporters may be involved in the pathophysiology of ICP.
Archives of Gynecology 12/2011; 285(6):1535-40. · 0.91 Impact Factor
[show abstract][hide abstract] ABSTRACT: The objective of this study was to determine maternal and placental concentrations of follistatin-like 3 (FSTL3), and, maternal concentrations of myostatin in gestational diabetes mellitus (GDM).
40 women with GDM of term pregnancy were recruited and 40 maternal age- and gestational age-matched normally pregnant women served as control. Maternal blood samples and placental tissues were collected. Maternal concentrations of FSTL3 and myostatin were determined by enzyme-linked immunosorbent assay (ELISA), and, placental concentrations of FSTL3 by Western blotting.
Women with GDM had significantly lower serum FSTL3 concentrations than controls (P=0.001). Placental concentrations of FSTL3 were significantly lower in GDM group than in controls (P<0.001). Women with GDM had significantly higher homeostasis model assessment of insulin resistance (HOMA-IR) and glycosylated hemoglobin (HbAlc) than control women (P=0.042 and <0.01, respectively). Maternal serum myostatin was not significantly different between GDM and control groups (P=0.312).
Maternal and placental FSTL3 concentrations were reduced in GDM women compared with normally pregnant women, suggesting FSTL3 may play an important role in the pathogenesis of gestational diabetes.
Clinica chimica acta; international journal of clinical chemistry 11/2011; 413(5-6):533-6. · 2.54 Impact Factor
[show abstract][hide abstract] ABSTRACT: Despite the efforts to recapitulate the follicle environment, oocytes from in vitro maturation (IVM) have poorer developmental potential than those matured in vivo and the effects on the resultant offspring are of concern. The aim of this study was to determine altered gene expression in oocytes following IVM and to evaluate the expression of the arginine rich, mutated in early stage of tumors gene (Armet) and mitochondrial ribosomal protein L51 (Mrpl51) in embryos and brains of fetal/postnatal mice and the brain development of IVM offspring. An IVM mouse model was established while oocytes matured in vivo were used as the controls. Suppressive subtractive hybridization (SSH) and RT-PCR/western blot were used to analyze the differential expression of genes/proteins between IVM and the control group. HE staining and water maze were used to assess the histological changes in brain tissue and cognition of the offspring. The rates of fertilization, cleavage, and live birth were significantly decreased in IVM group. Thirteen genes were upregulated in IVM oocytes compared with the control, including Armet and Mrpl51. The higher level of Armet in IVM oocytes was retained in brain of newborn mice, which could be related to the upregulation of activating transcription factor 6 (Atf6) and X-box binding protein 1 (Xbp1), while Mrpl51 was expressed normally in brain of postnatal mice. No significant differences were detected in brain weight, neuronal counts, and the cognition in the offspring between the two groups. The present results suggested that IVM could affect the pregnancy outcome and the Armet and Mrpl51 gene/protein expression. The change in Armet expression lasted while the change of Mrpl51 disappeared after birth. However, the brain development of the offspring seemed to be unaffected by IVM.
[show abstract][hide abstract] ABSTRACT: The aims were to clarify the effect of placental trophoblasts on T lymphocytes by assessing production of cytokines and expression of transcription factors regulating Th1, Th2, and Th17 immunity in T lymphocytes. Placental trophoblasts were isolated and conditioned medium was made after trophoblast cultivation for 72 h. T lymphocytes were cultured in presence or absence of conditioned medium. ELISA was used to detect concentration of IL-2, TNF-α, IFN-γ, IL-4, IL-10, and IL-17 in supernatants of T cell and real-time PCR was used to detect the status of Th1 (T-bet, STAT-4), Th2 (GATA-3, STAT-6), and Th17 (RORC) immunity in T lymphocyte. We found that the level of IL-2, IFN-γ, TNF-α, and IL-17 was significantly decreased when T lymphocytes were cultured in conditioned medium compared with control, while IL-10 and IL-4 level were not significantly changed. The presence of conditioned medium significantly decreased the ratio of Th1/Th2. The expression of GATA-3 and STAT-6 were significantly increased and STAT-4 was reduced when T lymphocyte was cultured in conditioned medium, while the expression of T-bet and RORC was not significantly different. We concluded that placental trophoblast-induced shift of Th1/Th2 balance toward Th2 and inhibition of Th17 might be among the mechanisms involved in maternal tolerance to fetus.
[show abstract][hide abstract] ABSTRACT: The purpose of the study was to clarify the alterations in serum leptin and soluble leptin receptor levels at term and early postpartum in gestational diabetes mellitus (GDM). Twenty women with normal pregnancy and 20 with GDM were recruited and blood samples were taken on the day of delivery and Days 1, 3 and 5 after delivery. Serum leptin levels were significantly higher in women with GDM than in the controls before delivery and decreased significantly after delivery (p < 0.001). After delivery there were no significant differences in serum leptin concentrations between women with GDM and the controls. Serum soluble leptin receptor concentrations did not differ neither between the two groups, nor before or after delivery. Leptin may play a role in GDM through a positive correlation with insulin resistance.
[show abstract][hide abstract] ABSTRACT: The present study sought to validate the gene chip technique to screen for gestational diabetes mellitus (GDM).
We selected 70 single-nucleotide polymorphisms (SNPs) associated with diabetes in previous studies, and SNP genotyping was performed in 130 cases (50 diabetes patients and 80 normal controls whose mean term of pregnancies are similarly 38-39 weeks). Relevant SNP patterns were established, and gene chips were designed after statistically analyzing the data. Peripheral blood samples were then collected from 24 healthy pregnant women and 24 pregnant women with GDM for gene chip analysis. The results of the gene chip method were also verified by DNA sequencing.
Four candidate SNPs (rs13266634, rs266729, rs3802177 and rs9300039) were obtained after genotyping. Primers were designed based on the four SNPs, and gene chips met the quality standard according to signal intensity. The genotypes of SNPs rs3802177, rs13266634 and rs266729 showed significant differences between healthy pregnant women and pregnant women with GDM, and these results were identical to those obtained with the DNA sequencing method.
We have demonstrated the feasibility of the gene chip technique in screening GDM and identified candidate loci with which to study this disease.
Diabetes research and clinical practice 08/2010; 89(2):167-73. · 2.16 Impact Factor
[show abstract][hide abstract] ABSTRACT: To investigate the alterations in gene profile of placenta from pregnant women with intrahepatic cholestasis of pregnancy (ICP) and to enhance the insight of etiology and pathogenesis of ICP.
Ten pregnant women diagnosed ICP were recruited and 10 healthy pregnant women served as control. Four samples were taken from each placenta and RNA was isolated. Gene expression was analyzed with microarray and real time PCR was used to validate the differentially expressed genes.
392 genes were found differentially expressed. Among these differentially expressed genes, 280 were up-regulated and 112 were down-regulated. These differentially expressed genes involved 20 categories including genes involved in transportation, cell growth, apoptosis and immune response that were putatively participated the pathogenesis of ICP.
293 differentially expressed genes of 20 categories were found in ICP placenta, suggesting the diversity of gene expression alteration and the complexity of etiology and pathogenesis of ICP.
Archives of Gynecology 07/2009; 281(5):801-10. · 0.91 Impact Factor
[show abstract][hide abstract] ABSTRACT: Preeclampsia, a pregnancy-specific complication occurring in the second half of human pregnancy and one of the leading causes for perinatal mortality and morbidity, is characterized by the onset of hypertension and proteinuria. We measured circulating adipocyte fatty acid binding protein (AFABP) and retinol binding protein-4 (RBP-4) in patients with preeclampsia.
Twenty-seven healthy non-pregnant women, 27 healthy pregnant women at third trimester and 26 women with preeclampsia were recruited and blood samples were taken. Concentrations of serum AFABP and RBP-4 were measured with ELISA.
There were significant differences in serum AFABP (median: 0.99, 0.93 and 1.81ng/ml, respectively, P<0.001) and RBP-4 (mean: 3.16, 2.65 and 4.27ng/ml, respectively, P=0.022) among non-pregnancy, normal pregnancy and preeclampsia. Serum AFABP was significantly higher in preeclampsia than non-pregnancy (P<0.001) and normal pregnancy (P=0.001). Serum RBP-4 was significantly higher in preeclampsia than normal pregnancy (P=0.007) but not non-pregnancy (P=0.061). There were no significant differences in serum RBP-4 and AFABP between non-pregnancy and normal pregnancy. Serum RBP-4 was significantly higher in severe than mild preeclampsia (mean: 5.15 vs 2.84ng/ml, P=0.046). There was no significant difference in serum AFABP between mild and severe preeclampsia.
Increased circulating AFABP and RBP-4 concentrations were demonstrated, suggesting it be an important pathophysiology of preeclampsia.
Clinica chimica acta; international journal of clinical chemistry 07/2009; 407(1-2):58-61. · 2.54 Impact Factor
[show abstract][hide abstract] ABSTRACT: To verify the hypothesis that vascular endothelial growth factor (VEGF) attenuates Nomega-Nitro-L-arginine Methyl Ester (L-NAME)-induced preeclampsia-like manifestations in rats.
Forty pregnant Wistar rats were randomly divided into four groups: control, preeclampsia model, VEGF treatment, and VEGF prophylactic. On day 5 of gestation, L-NAME was injected subcutaneously in rats of the preeclampsia model, VEGF treatment, and VEGF prophylactic groups. VEGF was given after the occurrence of hypertension and proteinuria in the VEGF treatment group and from day 5 in the VEGF prophylactic group. Blood pressure was monitored and urine protein was assayed. Blood platelet was counted, and serum nitric oxide metabolites, endothelin-1, 6-keto-PGF-1alpha, and TXB2 were determined.
Blood pressure increased significantly on day 8 of gestation in the preeclampsia model and VEGF treatment groups compared with control (p < 0.05 for both) and remained elevated through the pregnancy in the preeclampsia model group. Blood pressure was significantly decreased after the administration of VEGF in the VEGF treatment group (p < 0.05). There was no significant difference in blood pressure between the VEGF prophylactic group and control (p > 0.05). Urine protein, platelet count, serum nitric oxide metabolites, endothelin-1, 6-keto-PGF-1alpha, and TXB2 were significantly different between control and the preeclampsia model group (p < 0.05), but not between control and the VEGF treatment or VEGF prophylactic groups (p > 0.05 for all).
VEGF attenuates L-NAME-induced preeclampsia-like manifestations in rats, suggesting the important role of VEGF in preeclampsia and providing a potential strategy for the prevention and treatment of preeclampsia.
Clinical and Experimental Hypertension 10/2008; 30(7):606-15. · 1.28 Impact Factor
[show abstract][hide abstract] ABSTRACT: To clarify the change of peripheral CD4+CD25+ regulatory T lymphocytes in normal pregnancy and pre-eclampsia and to explore its role in the pathogenesis of pre-eclampsia.
Twenty-seven women with pre-eclampsia, 27 with normal third trimester pregnancy and 27 healthy non-pregnant women were recruited. Blood samples were taken and surface antigen CD4 and CD25 were labelled with fluorescence-conjugated antibodies. CD4+CD25+ regulatory T cells were analysed by flow cytometry, and the proportion in T cells and the amount of regulatory T lymphocytes were calculated.
The amount and the proportion of regulatory T cells were not significantly different among non-pregnant, normal pregnancy and pre-eclampsia groups (p>0.05 for both). There were no significant differences in the proportion and the amount of CD4+CD25+ regulatory T cells between mild and severe pre-eclampsia subgroups (p>0.05 for both).
No significant change of peripheral regulatory T cells was observed in the current investigation, suggesting other mechanisms rather than CD4+CD25+ regulatory T cells were involved in the pathogenesis of pre-eclampsia.
[show abstract][hide abstract] ABSTRACT: To characterise the changes in serum visfatin levels in late normal pregnancy and pre-eclampsia.
Twenty-seven women with pre-eclampsia were recruited. Twenty-eight women in the third trimester of normal pregnancy served as pregnant control and 28 healthy non-pregnant women as non-pregnant control. Serum levels of visfatin were measured with an enzyme-linked immunosorbent assay.
The means of serum visfatin were 626.4+/-45.5 ng/ml (mean+/-SEM) in non-pregnant control, 695.9+/-92.5 ng/ml in pregnant control, and 308.3+/-80.0 ng/ml in pre-eclampsia, respectively, and were significantly different among the groups (p<0.001). Visfatin level was significantly lower in pre-eclampsia compared to non-pregnant control (p=0.004) and pregnant control (p<0.001). Women with severe pre-eclampsia had a significantly lower serum visfatin level than those with mild pre-eclampsia (114.6+/-80.9 versus 425.2+/-122.1 ng/ml, p=0.037).
A decrease in visfatin level was demonstrated in pre-eclampsia, suggesting that visfatin and adipokine-associated metabolic abnormalities are involved in the pathogenesis of the disease.
[show abstract][hide abstract] ABSTRACT: Tolerance of T lymphocytes at the feto-maternal interface is necessary to sustain normal pregnancy. The present investigation aimed to observe the regulatory effects on T lymphocytes by human trophoblasts and to explore possible effector molecules.
Conditioned media was made by trophoblast culture or villous explant culture for T lymphocyte proliferation and proteomic analysis. Lymphocyte proliferation was tested by thymidine incorporation. Messenger RNA for indoleamine 2,3- dioxygenase (IDO) was detected by RT-PCR and tryptophan was assayed. The protein profile of conditioned media was assessed with shotgun mass-spectrometry and the identified proteins were bioinformatically analyzed. Human chorionic gonadotropin (HCG), human chorionic somatomammotropin (HCS), interleukin (IL)-2, 4, 10 and tumor necrosis factor (TNF)-alpha were assayed with radioimmunoassay (RIA).
T Lymphocyte proliferation was inhibited by conditioned medium in a dose-dependent manner. Inhibition of IDO during previous conditioning or addition of tryptophan to the conditioned medium partly restored T lymphocyte proliferation. mRNA for IDO was expressed in trophoblasts and chorionic villi. The concentrations of tryptophan were 19.01 and 3.79 micromol/L in unconditioned and conditioned media respectively. By proteomic procedures, 548 proteins were found in placenta-conditioned medium. Among these proteins were some proteins inhibiting T lymphocytes including HCG, HCS, AFP, pregnancy-specific beta 1-glycoprotein (SP1), glycodelin, transforming growth factor beta2, thrombospondin-1, pigment epithelium-derived factor (PEDF), galectin-1, and macrophage migration inhibitory factor. HCG and HCS were also detected with RIA, however, no interleukins were detected in conditioned media with RIA or proteomic analysis.
Trophoblasts inhibit T lymphocyte through IDO-mediated tryptophan depletion and placenta-derived immunoregulatory factors. Immunological tolerance at maternal-fetal interface represents a synergistic effect of these substances and a complex mechanism involving endocrine and immune networks.
Cellular Physiology and Biochemistry 02/2008; 21(5-6):463-72. · 3.42 Impact Factor
[show abstract][hide abstract] ABSTRACT: The objective was to investigate the alterations in estrogen receptor-alpha and -beta (ER-alpha and ER-beta) in the anterior vaginal wall of women with stress urinary incontinence (SUI).
Samples of the anterior vaginal wall were taken from 57 women, including 12 women with premenopausal SUI (pre-M SUI), 12 with premenopausal control (pre-M control), 19 with postmenopausal SUI (post-M SUI), and 14 with postmenopausal control (post-M control). The expressions of ER-alpha and ER-beta were assayed by immunohistochemistry and quantified with the H-score method.
Serum estradiol was significantly lower in the pre-M SUI than in the pre-M control group (P<0.01), but the difference between the post-M SUI and post-M control groups was not significant (P>0.05). ER-alpha in endothelia, smooth muscle cells, and fibrocytes were significantly lower in pre-M SUI than in pre-M control (P<0.01), but there were no significant differences of ER-alpha between the post-M SUI and post-M control groups (P>0.05). ER-beta in endothelia and fibrocytes were significantly lower in the pre-M SUI than in the pre-M control group (P<0.01), and ER-beta in fibrocytes was significantly lower in the post-M SUI than in the post-M control group (P<0.01).
Alterations in serum estradiol and its receptors (ER-alpha and ER-beta) in the anterior vaginal wall were demonstrated, suggesting their involvement in the occurrence of female SUI.
European Journal of Obstetrics & Gynecology and Reproductive Biology 11/2007; 134(2):254-8. · 1.84 Impact Factor
[show abstract][hide abstract] ABSTRACT: To observe the alterations in serum resistin in gestational diabetes (GDM) and the early postpartum period, and compare this to nondiabetic pregnancies in order to evaluate the role of serum resistin in gestational diabetes mellitus.
A cross-sectional study.
Twenty women with normal pregnancy and 20 with gestational diabetes.
Serum resistin concentration was assayed by ELISA.
Serum resistin concentration was significantly higher in women with GDM than in controls before delivery. Serum levels of resistin significantly decreased after delivery in both the GDM group and controls (P < 0.001 for both). There was a trend of higher serum resistin in women with GDM than in controls. Differences existed on days 1 (P < 0.001) and 3 (P = 0.013), but not by day 5 (P = 0.052) after delivery.
Elevated serum resistin level was observed in women with gestational diabetes, suggesting that it is important in the pathology of the disease.