Novel SNPs of WNK1 and AKR1C3 are associated with preeclampsia.

Preeclampsia is a hypertensive disorder of pregnancy and is one of the most common causes of poor perinatal outcomes. Preeclampsia increases the risk of hypertension in the future. Variants of WNK1 (lysine deficient protein kinase 1), ADRB2 (ß2 adrenergic receptor), NEDD4L (ubiquitin-protein ligase NEDD4-like), KLK1 (kallikrein 1) contribute to hypertension, and AKR1C3 (aldo-keto reductase family1 member C3), is associated with preeclampsia. The association of single nucleotide polymorphisms (SNPs) in these five candidate preeclampsia susceptibility genes and the related traits in Chinese individuals were investigated. In this study, 13 SNPs of the five genes were genotyped in 276 preeclampsia patients and 229 age- and area-matched normal pregnancies in women of Chinese Northern Han origin. The 95% confidence interval (CI) and odds ratio (OR) were estimated by binary logistic regression. No obvious linkage disequilibrium or haplotypes were observed among these SNPs. Those with GG genotype and allele G of AKR1C3 (rs10508293) had a decreased risk of preeclampsia (adjusted OR = 3.011, 95% CI = 1.758-5.159, and adjusted OR = 1.745, 95% CI = 1.349-2.257, respectively). The AA genotype and allele A of WNK1 (rs1468326) were significantly associated with an increased risk in preeclampsia (adjusted OR = 2.307, 95% CI = 1.206-3.443, and adjusted OR = 1.663, 95% CI = 1.283-2.157, respectively). The findings indicate that the GG genotype of AKR1C3 rs10508293 is associated with decreased risk for preeclampsia and the AA genotype of WNK1 rs1468326 are related with an increased risk for preeclampsia.

Associations of polymorphisms of CYP2D6 and CYP2C9 with early onset severe pre-eclampsia and response to labetalol therapy.

PURPOSE: Early onset preeclampsia (PPE) contributes to life-threatening maternal complications and fetal demise. Pharmacogenomics is a precision medicine, and metabolizing enzymes responsive to antihypertensive remains understudied. The aim of this study was to evaluate the associations of polymorphisms of cytochrome P450, family 2, subfamily D, polypeptide 6 (CYP2D6) and cytochrome P450, family 2, subfamily C, polypeptide 9 (CYP2C9) with PPE and the relationship among CYP2D6, CYP2C9 polymorphisms and response to labetalol therapy. METHODS: Totally 105 gravidas diagnosed with PPE (case) and 103 healthy gravidas (control) were recruited between August 2013 and July 2016. Labetalol was given to control blood pressures (BP) with PPE. If labetalol administration alone did not exceed the mean dose and effectively controlled the BP, it would be considered to be valid (n = 75). Genotype and allele frequencies of CYP2C9 gene (rs1057910 and rs4918758) and CYP2D6 gene (rs1065852, rs28371725, rs35742686, and rs3892097) were analyzed by TaqMan PCR. Differences in the genotype and allele frequencies were compared between case-control groups, and the responsive and nonresponsive to labetalol in PPE. RESULTS: Out of six variants, only CC and CT genotypes of the CYP2D6 variants (rs28371725) in PPE were significantly higher than those in the control group [18.1% (19/105) vs 14.6% (15/103); 56.2% (59/105) vs 42.7% (44/103); χ2 = 6.707]. However, there were no differences in maternal age, diastolic pressure, BMI, BW, serum triglyceride, and creatinine were observed among women with CC, CT, or TT genotype of CYP2D6 gene rs28371725 in the experimental group (all P > 0.05). Compared with the gravidas with CT or TT genotype of CYP2D6 gene rs28371725, those with CC genotype had longer gestational age [(32.5 ± 2.1) vs (29.5 ± 1.8) and (29.8 ± 2.2) weeks] and higher plasma albumin [(27.2 ± 9.3) vs (20.3 ± 10.4) and (22.5 ± 7.4) g/L], but lower systolic pressure and 24 h urine protein (LSD test, all P < 0.05). The G allele frequency in CYP2D6 gene rs1065852 nonresponsive to labetalol group was higher than that in responsive labetalol group [93.3% (56/60) vs 76.0% (114/150), χ2 = 8.351, P = 0.004]. CONCLUSIONS: The polymorphism of CYP2D6 gene rs28371725 may be associated with PPE, and the allele of G in CYP2D6 gene rs1065852 may be associated with the efficacy of labetalol in treatment of PPE.

Role of AKR1C3 in renal injury and glibenclamide is anti-inflammatory in preeclamptic rats.

Proteinuria is a common adverse effect of renal injury in preeclampsia. To explore the effects of AKR1C3 in renal injury due to preeclampsia and to determine an optimal method to prevent proteinuria, glibenclamide treatment was used in unrestrained Wistar rats exposed to N-nitro-L-arginine methyl ester hydrochloride (L-NAME). Successful rat models for preeclampsia were confirmed based on mean arterial pressure, a 24-h protein urine test, and by observing the structure of the kidney by transmission electron microscopy (TEM). Forty Wistar rats were randomly divided into L-NAME-induced preeclampsia (pregnant and L-NAME), treatment (pregnant, L-NAME, and glybenclamide), non-pregnant (L-NAME), and control (pregnant and 0.9% saline) groups. Rats that were 19 days into their pregnancies were then euthanized and their kidneys were collected. After exposure to L-NAME, the mean arterial pressure increased by ~25 mmHg, which was largely prevented by the co-administration of glibenclamide. At 24 h, protein levels in the urine of the L-NAME-induced preeclampsia group were higher than those of the control and treatment groups. AKR1C3 was downregulated in the kidney and podocytes, whereas glibenclamide increased the expression of AKR1C3. The generation of reactive oxygen species (ROS) detected by ELISA was decreased by the glibenclamide co-administration. Compared with that in the L-NAME-induced preeclampsia group, the expression levels of AKR1C3 protein and mRNA significantly increased in the treatment group ([0.48 ±â€¯0.09] vs.[1.05 ±â€¯0.20];[0.05 ±â€¯0.02] vs.[0.22 ±â€¯0.06]; both P < 0.05]). Therefore, AKR1C3 expression was decreased in the kidneys of L-NAME-induced preeclampsia rats, and glibenclamide may be useful for the treatment of preeclampsia by regulating the generation of ROS and preventing proteinuria.

Twin pregnancy and partial hydatidiform mole following in vitro fertilization and embryos transfer: a novel case of placental mosaicism.

Twin pregnancy with mosaic partial hydatidiform mole (PHM) and survival of two healthy fetuses following in vitro fertilization and embryos transfer (IVF-ET) is a rare situation and is considered a challenge for management. A 32-year-old Chinese woman conceived twin pregnancy following IVF-ET. At 22 weeks' gestation, an additional intrauterine echogenic mass with features of PHM were shown by successive ultrasound examinations. At 35 weeks' gestation, two live male infants and two placentas were delivered by caesarean section (CS). Histologic examination of the abnormal placenta confirmed mosaic PHM. Genetic study showed the abnormal placental mosaicism (expressed in molar-69XXY and normal vili-46XY), co-existing with a hypospadia new-born (46XY) in one amniotic sac. However, the other one was normal. Serial serum ß-hCG levels showed a declining trend and serum ß-human chorionic gonadotropin (hCG) were undetectable at 6 months after delivery. The case demonstrated that it is possible to prolonged gestation by PHM under close surveillance during the entire pregnancy.

[Screen and identify of differential proteins expressed in the placenta of Down's syndrome].

OBJECTIVE: To discuss protein marks expressed differentially in placenta of Down's syndrome by means of proteomics. METHODS: We collected placenta of 18 patients (from March 2009 to December 2009 at Beijing Obstetrics and Gynecology Hospital), and divided them into two groups, one was 10 patients with fetal Down's syndrome, the other was normal pregnancies (normal chromosome) with other diseases. We separated proteins expressed in placentas of two groups by two-dimensional difference gel electrophoresis (2D-DIGE), and then analyzed the differential protein spots by software Decyder 6.5, then, spots differentially expressed by 1.5 fold or more were analyzed by matrix assisted laser desorption ionization-time of flight-mass spectrometry (MALDI-TOF-MS). In the end, the differential expressional levels of partially identified proteins were validated by western blot analysis. RESULTS: (1) Differential proteins of two groups protein spots of placentas separated by 2D-DIGE were analyzed by software Decyder 6.5 (these colored lights scattered in the image were protein spots), a total of 56 spots out of 352 were differentially expressed (P < 0.05) in two groups. We analyzed 17 protein spots (12 protein spots were over-expressed and 5 protein spots were down-expressed) differentially expressed by 1.5 fold or more by MALDI-TOF-MS. (2) Protein matching after searching protein database, 17 protein spots turn out to be 10 proteins. Four kinds [superoxide dismutase 1 (SOD1), peroxiredoxin 6 (PRDX6), heat shock protein 27 (HSP27), endoplasmic reticulum protein 29 (ERP29)] of them were validated by western blot analysis, the group of fetal Down's syndrome were 0.74 ± 0.12, 0.29 ± 0.10, 0.53 ± 0.16, 0.20 ± 0.09, the group of normal pregnancies were 0.51 ± 0.08, 0.34 ± 0.16, 0.18 ± 0.07, 0.35 ± 0.09, the results confirmed the observed changes in proteomics. CONCLUSIONS: Compared with normal pregnancies, there were differential proteins expressed in placenta of Down's syndrome. This approach might provide new screening markers in use for prediction of Down's syndrome, however, further study should be done to make these 4 proteins (SOD1, HSP27, ERP29, PRDX6) be new screening markers.

Genetic polymorphisms of killer cell immunoglobulin-like receptor 3DL2 in preeclampsia.

AIMS: We evaluated if killer cell immunoglobulin-like receptor 3DL2 gene (KIR3DL2) polymorphisms are a key factor in the development of preeclampsia. METHODS: In this case-control study, 105 pregnant women with PE (PE group) were enrolled. Their A52G in exon 3 and C32T in exon 9 polymorphisms of the KIR3DL2 genotypes were determined by polymorphism chain reaction-restriction fragment length polymorphism (PCR-RFLP) from venous blood samples and compared with the corresponding KIR3DL2 genotypes of 103 pregnant women with uncomplicated pregnancies (control group). RESULTS: Carriers of the A allele in exon 3 of KIR3DL2 gene occurred less frequently in PE than in controls [P=0.001; odds ratio (OR)=2.65, range: 1.5-4.7]. No significant difference was found about allelic frequencies of KIR3DL2 gene C32T in exon 9 in women with preeclampsia as compared to controls. A significant difference between the two groups of genotypic frequencies of KIR3DL2 gene A52G in exon 3 and KIR3DL2 gene C32T in exon 9 polymorphisms was found (P=0.003 and P=0.000). There was no significant difference between genotypic or allelic frequencies in women with mild preeclampsia compared to sever preeclampsia. CONCLUSIONS: Our results suggest that carriers of A allele in exon 3 have a decreased susceptibility to PE. It is likely that the presence of the CC genotype in exon 9 has a considerable effect on disease progression. The mutation of the two sites is not associated with the severity of preeclampsia.

Proteomic analysis of the alteration of protein expression in the placenta of Down syndrome.

BACKGROUND: Down syndrome (DS) is the most common form of human aneuploidy, and there is no effective therapy for the chromosomal abnormalities. We aimed to unravel the molecular mechanisms underlying DS and to provide clues to prenatal screening. METHODS: A series of proteomics-based experiments was conducted using 19 patients with DS fetuses and 17 normal pregnancies. The proteome of placenta was investigated as displayed by two-dimensional difference gel electrophoresis (2D-DIGE), and comparisons were made between placentas that developed under DS and normal pregnancy conditions. Multivariate analysis of the resulting protein patterns revealed DS-specific protein expression. Matrix-assisted laser desorption/ionization (MALDI) time-of-flight/time-of-flight (TOF/TOF) high-resolution tandem mass spectrometer (MS)-based identification was successful for 12 out of 17 selected protein spots. RESULTS: Among those, three proteins involved in the resist of reactive oxygen species (ROS) and neurogenesis were more abundant in the DS placenta (superoxide dismutase 1, endoplasmic reticulum protein 29 and heat shock protein beta-1), while peroxiredoxin-6 involved in cell defense mechanism against ROS was expressed at a higher level in the normal pregnancies. CONCLUSION: Knowledge of the DS placenta proteome emphasizes the role of proteins involved in anti-oxidation during DS, and may form the basis of a potential approach to minimize the incidence of DS in the clinical setting.

[Clinical characteristics and perinatal outcomes of non-overweight/obese pregnant women with polycystic ovary syndrome].

OBJECTIVE: To determine clinical characteristics and perinatal outcomes of non-overweight/obese (pre-pregnancy body mass index BMI < 24 kg/m(2)) pregnant women with polycystic ovary syndrome (PCOS). METHODS: The screening of PCOS was performed when they were at first prenatal visit in Beijing Obstetrics and Gynecology Hospital, Capital Medical University from May, 2008 to July, 2010.61 PCOS women of pregnancy women with BMI < 24 kg/m(2) were considered as the study subjects, and 122 pregnant women without PCOS matched by age and pre-pregnancy body mass index (BMI) were selected as the control ones. Patients with history of pre-pregnancy diabetes, hypertension, cardiovascular disease, renal diseases and multiple pregnancies were excluded from the study. We followed pregnancy outcomes of two groups until delivery. RESULTS: Significantly higher total cholesterol, triglycerides concentrations and low-density lipoprotein cholesterol concentrations were found in the PCOS group than in the controls. PCOS women with BMI < 24 kg/m(2) had significantly higher rates of GDM (27.9% (17/61)) and pre-eclampsia (13.1% (8/61)) compared with the controls (15.6% (19/122), 1.6% (2/122)), P < 0.05, < 0.01, respectively. No statistical significance was found in prevalence of pregnancy-induced hypertension, polyhydramnios, oligohydramnios, macrosomia, premature rupture of membranes, placental abruption, macrosomia, fetal death and neonatal congenital abnormality between the two groups (all P > 0.05). CONCLUSIONS: It is at increased risk of GDM and pre-eclampsia in non-overweight/obese PCOS women, this risk seemed to be due to PCOS itself rather than obesity.

[Study of follicle-stimulating hormone receptor and tyrosine hydroxylase polymorphisms and pre-eclampsia in Chinese Han population].

OBJECTIVE: To analyze a potential association of gene polymorphisms of the follicle-stimulating hormone receptor (FSHR) gene rs1394205 polymorphism and tyrosine hydroxylase (TH) rs2070762 polymorphism with pre-eclampsia. METHODS: The gene polymorphism of FSHR rs1394205 and TH rs2070762 were analyzed by real-time Quantitative polymerase chain reaction (TaqMan probe) in 105 patients with pre-eclampsia and 103 healthy pregnant subjects in Chinese Han population. Genotype and allele were assessed by direct counting methods. And the associations between 2 SNPs and pre-eclampsia were analyzed by chi-square test. RESULTS: The frequencies of the TH gene rs2070762 polymorphism CC, C/T and TT genotypes were 14.6%, 42.7% and 42.7% in pre-eclampsia group and 18.1%, 56.2% and 25.7% in normal control group respectively. The allelic frequency of rs2070762 polymorphism in TH in pre-eclampsia patients was significantly lower (P < 0.05) than that in normal pregnancies. There was a significant difference between two groups (P < 0.05). But there was no significant difference between mild and severe pre-eclampsia groups (P > 0.05). In contrast, no significant association was detected in the comparison of genotypes and allele of FSHR gene rs1394205 between pre-eclampsia patients and normal pregnancies. CONCLUSION: The TH rs2070762 polymorphism was associated with the pathogenesis of pre-eclampsia. It suggests that the polymorphism in TH may be involved in the onset and development of pre-eclampsia.

G protein-coupled receptor kinase 4 gene variants are not associated with preeclampsia in Northern Han Chinese.

Genetic variations in preeclampsia (PE) may affect PE risk. The G protein-coupled receptor kinase 4 (GRK4) gene encodes a member of the Ser/Thr protein kinase family and has been linked to both genetic and acquired hypertension. The aim of this study was to investigate the association between polymorphisms (T-rs1024323-C and T-rs1801058-C) in GRK4 and PE in Northern Han Chinese. Using a case-control design, the association between the GRK4 exon-4 T-rs1024323-C and exon-13 T-rs1801058-C polymorphisms and the risk of PE in Northern Han Chinese was assessed in 105 individuals with PE and 103 age- and area-matched normotensive controls. Genotypes were determined by allelic discrimination. The odds ratio and 95% confidence interval were estimated by binary logistic regression. No association was found between the GRK4 polymorphisms (T-rs1024323-C and T-rs1801058-C) and PE, and there was also no relationship with the severity of PE. The risk of homozygous and heterozygous variant allele carriers of the analyzed single-nucleotide polymorphisms did not differ significantly from that of the homozygous wild-type allele carriers, even after adjustment for age, body mass index, (family) history of hypertension and smoking status. The GRK4 (T-rs1024323-C and T-rs1801058-C) polymorphisms were not associated with a risk of PE in the present Northern Han Chinese study group. Thus, the GRK4 polymorphisms do not seem to have an important role in PE in this population.

[The study of killer cell immunoglobulin-like receptor 3DL2 gene polymorphisms and pre-eclampsia].

OBJECTIVE: To analyze a presumed association of gene polymorphism of killer cell immunoglobulin-like receptor three domains, long cytoplasmic tail, 2(KIR3DL2) gene with pre-eclampsia. METHODS: The KIR3DL2 gene A52G in exon 3 and C32T in exon 9 polymorphism were determined by polymerase chain reaction-restricted fragment length polymorphism (PCR-RFLP) in 95 pre-eclampsia and 91 normal pregnant women. RESULTS: There was significant difference between two group in exon 3 in terms of genotypes and allele (P < 0.05). There was a significant difference between two groups regarding the frequencies of exon 9 CC, CT and TT genotypes (P < 0.05). But no significant difference was found between mild pre-eclampsia and severe pre-eclampsia groups in terms of the two SNP loci (P > 0.05). CONCLUSION: The A52G polymorphism in exon 3 and the frequencies of C32T in exon 9 of KIR3DL2 gene polymorphism are associated with the pathogenesis of pre-eclampsia, But it has no correlation with the severity of preeclampsia.

Gene expression profiling of maternal blood in early onset severe preeclampsia: identification of novel biomarkers.

AIMS: To investigate candidate genes in peripheral blood mononuclear cell (PBMC) that are associated with early onset severe preeclampsia (ES-PE) and to describe candidate genes function using microarrays and real-time polymerase chain reaction (PCR). METHODS: PBMC RNA was extracted from six patients with ES-PE and five uncomplicated pregnancies. The HG_U133 plus 2.0 Affymetrix GeneChips that represented 47,000 genes were used to measure gene expression in each sample. Significance analysis of microarray identified potential signature genes characterizing ES-PE vs. uncomplicated pregnancies. Eight genes were selected for confirmation by real-time PCR of 32 patients with ES-PE and 24 uncomplicated pregnancies, matched for maternal age, parity, race and gestational weeks. RESULTS: Using a whole-genome approach to study the molecular determinants of ES-PE, 72 genes were found to be differentially expressed between cases and controls, including 38 up-regulated genes and 34 down-regulated genes in the group of ES-PE. Killer cell immunoglobulin-like receptor, three domains, long cytoplasmic tail, 2 (KIR3DL2), aldo-keto reductase family 1, member C3 (AKR1C3), churchill domain containing 1 (CHURC1), and solute carrier family 25, member 13 (SLC25A13) were validated to be down-regulated in the patients with ES-PE by real-time PCR. CONCLUSIONS: Expression of genes with diverse function is associated with ES-PE risk, providing opportunities for the development of non-invasive diagnosis.

[Microarray analysis of differentially expressed genes in peripheral leucocytes derived from severe preeclampsia and normotensive pregnancies].

OBJECTIVE: To investigate genes involved in the mechanisms underlying the progression of severe preeclampsia. METHODS: We conducted a multiregional gene expression analysis using peripheral leucocytes from patients with preeclampsia and normal controls. Total RNA was extracted from peripheral blood of six severe preeclampsia and five normotensive pregnancies. We performed genome-wide expression profiling using Affymetrix HG_U133 plus 2.0 chips to screen out differentially expressed genes of 2 fold or more and q_value < 5.4%. Using Gene Ontology we identified the function of differentially expressed genes after cluster analysis. RESULTS: Among the 47 000 genes that were screened in the microarray, 140 genes were found to be differentially expressed between normal and preeclamptic pregnancies. Eighty six up-regulated candidate genes were mainly involved in cysteine metabolism urea cycle and metabolism of amiogroups, proteasome, TGF-beta signaling pathway, and the ratio of calponin2 (CNN2), matrix metallopeptidase 8 (MMP8), V-set and immunoglobulin domain containing 4 (VSIG4), proteasome 26S subunit ATPase 5 (PSMC5) was evidently increased in preeclampsia patients. Among 54 down-regulated candidates, natural killer cell mediated cytotoxicity, antigen processing and presentation, metabolism of xenobiotics by cytochrome P450 were the main pathways. KIR3DL2, AKR1C3, CHURC1 and SLC25A13 were obviously decreased in preeclampsia patients. CONCLUSIONS: The gene expression of peripheral leucocytes in preeclampsia patients is significantly different from that of uncomplicated pregnancies. CNN2, MMP8, VSIG4, PSMC5, KIR3DL2, AKR1C3, CHURC1 and SLC25A13 may be involved in the molecular mechanisms underlying the progression of severe preeclampsia.