OBJECTIVE: To investigate the effect of vitamin D on microRNA-21(miR-21) expression and migration and invasion of human placental trophoblast cells. METHODS: The changes in the expression of miR-21 were detected using RT-qPCR in HTR-8/SVneo cells following stimulation by vitamin D at different doses for 24, 48 and 72 h.HTR-8/SVneo cells transfected with miR-21 mimic or inhibitor with or without vitamin D treatment were examined for changes in cell migration and invasion abilities using Transwell assay, and Western blotting was used to detect protein expressions of E-cadherin, fibronectin, and MMP9. RESULTS: Vitamin D obviously inhibited the expression of micoRNA-21 in HTR-8/SVneo cells in a concentration-and time-dependent manner.Transfection with the miR-21 mimic significantly inhibited the migration and invasion of HTR-8/SVneo cells, and this inhibitory effect was abolished by treatment with vitamin D; transfection with miR-21 inhibitor obviously promoted the migration and invasion of HTR-8/SVneo cells, and these effects were not significantly affected by vitamin D treatment. CONCLUSIONS Vitamin D may promote trophoblast cell migration and invasion to accelerate the development of preeclampsia by down-regulating the expression of miR-21.
Cell Movement ; Female ; Humans ; MicroRNAs ; genetics ; Placenta ; Pre-Eclampsia ; Pregnancy ; Trophoblasts ; Vitamin D

To investigate the expressions of placental growth factor (PLGF) in placenta with hypertensive disorders of pregnancy (HDP), 45 women with HDP and 20 normally pregnant women were studied. Among 45 women with HDP, there were 23 cases of severe preeclampsia and one case of eclampsia. The location and level of PLGF proteins was determined by immunohistochemistry and Western blot. The expression of PLGF mRNA in placenta was assessed by reverse transcriptional-polymerase chain reaction (RT-PCR). The results showed that: (1) The distribution of PLGF in placenta with HDP was similar to normal one, which was mainly in the cytoplasm of villous syncytiotrophoblast and villous stroma; (2) The expression of PLGF protein was significantly decreased in placentas with mild and severe preeclampsia compared to the normal ones (0.3 +/- 0.4 vs 0.6 +/- 0.4, 0.2 +/- 0.5 vs 0.6 +/- 0.4, P < 0.01). There were no differences between the gestational hypertension placenta and normal one (0.5 +/- 0.6 vs 0.6 +/- 0.4, P > 0.05); (3) The transcription levels of the PLGF mRNA in placentas with preeclampsia were significantly lower than in normal groups (3.33 +/- 0.39 vs 4.87 +/- 0.60, 1.97 +/- 0.29 vs 4.87 +/- 0.60, P < 0.01), and no differences were found between the gestational hypertension placenta and normal groups. These findings suggest that the abnormal expression of PLGF in placentas is related to the pathogenesis of HDP.
Placenta/*metabolism ; Pre-Eclampsia/*metabolism ; Pregnancy/*metabolism ; Pregnancy Proteins/*biosynthesis ; Pregnancy Proteins/genetics

We tested the hypothesis that angiotensin-converting enzyme (ACE) and angiotensinogen gene polymorphism influence the incidence, development and outcome of preeclampsia. Subjects were recruited from 90 Korean patients with preeclampsia during pregnancy and 98 age-matched controls. After isolation of DNA, polymerase chain reactions (PCR) were carried out to detect polymorphism of the ACE and angiotensinogen. M235T and T174M genotypes of angiotensinogen were determined by digestion with restriction enzyme endonuclease Tth 111-I and NCo I, respectively. The frequency of DD genotype was significantly greater in preeclampsia (0.36) than in controls (0.14) (p<0.05). The frequency of D allele was 0.55 in preeclampsia and 0.40 in controls (p<0.05). There were no differences in the onset of preeclampsia and pregnancy outcomes according to the ACE genotypes. There was no difference in the frequency of a allele of angiotensinogen M235T between the groups (0.79:0.78 in preeclampsia : controls). The frequency of T allele of angiotensinogen T174M gene was slightly increased, but not significantly, in preeclampsia (0.11) than in controls (0.07). In a multivariate analysis, only ACE genotype was associated with the development of preeclampsia (beta=0.27, p=0.05). In conclusion, a molecular variant of ACE, but not angiotensinogen, gene is associated with preeclampsia in Korean women.
Adult ; Angiotensinogen/*genetics ; Female ; Gene Frequency ; Genotype ; Human ; Korea ; Peptidyl-Dipeptidase A/*genetics ; *Polymorphism (Genetics) ; Pre-Eclampsia/*genetics ; Pregnancy

The expression of endothelial nitric oxide synthase traffic inducer (NOSTRIN) was examined in the umbilical vessels of the patients with pre-eclampsia (PE) to explore its possible role in the pathogenesis of PE. The NOSTRIN mRNA in umbilical tissues was determined by RT-PCR. The eNOS activity in umbilical vessels was spectrophotometrically detected. NO2-/NO3-, the stable metabolic end products of NO, was measured by using nitrate reductase. RT-PCR showed that the expression level of NOSTRIN was significantly higher in women with PE than in the normal group (P<0.01). The activity of eNOS was significantly decreased in PE group [(12.83+/-3.61) U/mg] than in normal group [(21.72+/-3.83) U/mg] (P<0.01). The level of NO2-/NO3- in PE patients (27.53+/-7.48) micromol/mg was significantly lower than that of normal group (54.27+/-9.53) micromol/mg (P<0.01). The significant negative correlation existed between the expression of NOSTRIN and the activity of eNOS in umbilical vessels of women with PE (r=-0.58, P<0.01). It was concluded that the level of NOSTRIN expression was increased in umbilical vessel of women with PE, indicating that it may be involved in the pathogenesis of PE.
Intracellular Signaling Peptides and Proteins/genetics ; Intracellular Signaling Peptides and Proteins/*metabolism ; Pre-Eclampsia/*enzymology ; Pre-Eclampsia/etiology ; RNA, Messenger/genetics ; RNA, Messenger/metabolism ; Umbilical Arteries/cytology ; Umbilical Arteries/*enzymology ; Umbilical Veins/cytology ; Umbilical Veins/*enzymology

PURPOSE: Recently, COMMD1 has been identified as a novel interactor and regulator of hypoxia-inducible factor-1 and nuclear factor kappa B transcriptional activity. The goal of this study was to determine the difference of COMMD1 expression in the placentas of women with normal and preeclamptic (PE) pregnancies. MATERIALS AND METHODS: Immnoperoxidase and immunofluorescent staining for COMMD1 was performed on nine normal and nine severe PE placental tissues, and COMMD1 mRNA expression was quantified by quantitative reverse transcription polymerase chain reaction. RESULTS: The expression of mRNA of COMMD1 was significantly higher in the study group than in the control group. The immunoreactivity was higher especially in the syncytiotrophoblast of PE placentas than in the control group. CONCLUSION: This study demonstrated increased placental COMMD1 expression in women with severe preeclampsia compared to that found in women with normal pregnancies, and this finding might contribute to a better understanding of the pathophysiology of preeclampsia.
Adaptor Proteins, Signal Transducing/genetics/isolation & purification/*metabolism ; Adult ; Female ; Humans ; Placenta/*metabolism ; Pre-Eclampsia/*metabolism ; Pregnancy ; RNA, Messenger/metabolism

OBJECTIVETo investigate the expression of annexin V in the decidua tissues of preeclampsia patients and explore its clinical significance.

METHODSReal-time PCR, Western blotting and immunohistochemistry were employed to detect the mRNA and protein expressions of annexin V in the deciduas from 35 normal pregnant women at full term, 38 early onset severe preeclampsia patients and 33 late onset severe preeclampsia patients.

RESULTSAnnexin V was found on the cell membrane and in the cytoplasm of the decidual cells and stroma. Both the mRNA and protein of annexin V expressions in the decidua tissues were significantly different between normal pregnancy group and early or late onset severe preeclampsia group (P<0.05), being the highest in normal pregnancy group and the lowest in early onset severe preeclampsia group.

CONCLUSIONThe low expression of annexin V in the deciduas might participate in the hypercoagulability state in preeclampsia patients.

Adult ; Annexin A5 ; metabolism ; Case-Control Studies ; Decidua ; metabolism ; Female ; Humans ; Immunohistochemistry ; Pre-Eclampsia ; metabolism ; pathology ; Pregnancy ; RNA, Messenger ; genetics

BACKGROUNDPreeclampsia, characterized by hypertension and proteinuria, is a multifactorial disease associated with shallow invasion of trophoblast cells and inadequate spiral artery remodeling. Trophoblast and tumor cells have similar invasion mechanism. Prostasin is closely related to tumor development, invasion and metastasis and influences blood pressure through activating epithelial sodium channel. The effect of prostasin on the pathogenesis of preeclampsia remains unclear. This study investigated the association of prostasin gene at rs12597511 with severe preeclampsia.

METHODSA single nucleotide polymorphism, rs12597511, was tested with polymerase chain reaction and restrictionfragment length polymorphism analyses in 179 severe preeclampsia patients and 222 normal pregnant women.

RESULTSThe frequencies of TC + CC genotypes were significantly higher in severe preeclampsia group compared with in control group (the adjusted odds ratio was 2.030, 95% confidence interval 1.195-3.449, P = 0.009). The C allele of rs12597511 was present significantly more often among women with severe preeclampsia (P = 0.001). Genotyping analysis showed that the C allele of rs12597511 could confer a risk for severe preeclampsia.

CONCLUSIONThe higher frequency of C allele of prostasin gene at rs12597511 is associated with severe preeclampsia.

Adult ; Female ; Gene Frequency ; genetics ; Genetic Predisposition to Disease ; genetics ; Genotype ; Humans ; Middle Aged ; Polymorphism, Single Nucleotide ; genetics ; Pre-Eclampsia ; genetics ; Pregnancy ; Serine Endopeptidases ; genetics ; Young Adult

OBJECTIVETo investigate the differential expression of adrenomedullin (ADM) in the placentas of women with normal and preeclamptic pregnancies, and explore the importance of ADM and its signal pathway in the development of preeclampsia.

METHODSTen pregnant women complicated with preeclampsia during the late term(>or=35 wk) were selected for this study along with 7 normal control pregnant women (>or=39 wk). The total RNA was extracted and sections of fresh placental tissues were prepared, and ADM expressions at mRNA and protein levels in women with normal and preeclamptic pregnancies were determined by Northern blotting and immunohistochemistry, respectively.

RESULT AND CONCLUSIONAt both mRNA and protein levels, the expression of ADM in the placentas of preeclamptic women was significantly reduced obviously as compared with that of the normal control (P<0.05), suggesting that ADM expression reduction in preeclamptic placenta might be associated with the development of preeclampsia.

Adrenomedullin ; biosynthesis ; genetics ; Adult ; Blotting, Northern ; Female ; Gene Expression Profiling ; Humans ; Immunohistochemistry ; Placenta ; metabolism ; Pre-Eclampsia ; genetics ; metabolism ; Pregnancy ; RNA, Messenger ; biosynthesis ; genetics

BACKGROUNDPreeclampsia (PE) is a multifactorial pregnancy complication. Maternal underlying condition and adverse factors both influence the pathogenesis of PE. Abnormal lipid metabolism as a maternal underlying disease may participate in the occurrence and development of PE. This study aimed to observe the effects of adverse factors on PE-like symptoms of pregnant mice with genetic abnormal lipid metabolism.

METHODSApolipoprotein C-III (ApoC3) transgenic mice with abnormal lipid metabolism were subcutaneously injected with L-arginine methyl ester (L-NAME) or normal saline (NS) daily starting at Day 7 or 16 of pregnancy (ApoC3+L-NA and ApoC3+NS groups), and wild-type (WT) mice served as a control (WT+L-NA and WT+NS groups). All mice were subdivided into early and late subgroups by injection time. The mean arterial pressure (MAP) and urinary protein were measured. Pregnancy outcomes, including fetal weight, placental weight, live birth rate, and fetal absorption rate, were analyzed. Pathologic changes in the placenta were observed by hematoxylin-eosin staining. One-way analysis of variance, t-test, and χ(2) test were used for statistical analysis.

RESULTSMAP significantly increased for ApoC3+NS groups compared with WT+NS groups (P < 0.05), without significant difference in urine protein. Following L-NAME injection, MAP and urinary protein significantly increased for ApoC3+L-NA and WT+L-NA compared with the corresponding NS groups (P < 0.05), and the increase for ApoC3+L-NA was more obvious. Urinary protein levels in early ApoC3+L-NA and WT+L-NA significantly increased compared with the corresponding late groups (P < 0.05). Fetal absorption rate significantly increased and fetal and placental weights significantly decreased in early ApoC3+L-NA and WT+L-NA compared with the corresponding NS groups (P < 0.05), without significant difference in late ApoC3+L-NA and WT+L-NA groups. Fetal weight in early ApoC3+L-NA was significantly lower than in early WT+L-NA group (P < 0.05). Morphologic examination of placentas from early ApoC3+L-NA and WT+L-NA groups showed varying degrees of fibrinoid necrosis.

CONCLUSIONSApoC3 transgenic mice with abnormal lipid metabolism showed gestational hypertension. Adverse factors and early effect time could aggravate the PE-like symptoms for ApoC3 transgenic mice.

Animals ; Apolipoprotein C-III ; genetics ; metabolism ; Female ; Lipid Metabolism ; drug effects ; genetics ; Male ; Mice ; Mice, Transgenic ; NG-Nitroarginine Methyl Ester ; pharmacology ; Pre-Eclampsia ; genetics ; metabolism ; Pregnancy

OBJECTIVETo assess the association of prostasin gene rs12597511 polymorphism with clinical features and pregnancy outcomes among patients with severe preeclampsia.

METHODSClinical manifestations, pregnancy outcomes and the genotypes of 179 patients with severe preeclampsia [early-onset group (≤34 gestational weeks): 79 cases; Late-onset group (>34 gestational weeks): 100 cases] and 222 normal-term pregnant women (control group) were collected.

RESULTSIn the early-onset group, the patients with TC or CC genotype at rs12597511 had higher incidences of total complications, liver dysfunction, neonatal asphyxia, neonatal intracranial hemorrhage and perinatal mortality compared with those with TT genotype (P>0.05). Multiple logistic regression analysis showed that the complication rates of severe preeclampsia patients are closely related to TC or CC genotypes, 24 h urinary protein and gestational weeks of onset (OR=1.049, 95% CI:1.007-1.093, P=0.021; OR=1.031, 95% CI: 0.350-0.883, P=0.013; OR=0.733, 95% CI: 0.566-0.950, P=0.019), and the perinatal mortality is related to gestational weeks at delivery (OR=0.542, 95% CI: 0.331-0.887, P=0.015).

CONCLUSIONPolymorphism of the prostasin gene is closely associated with poor pregnancy outcomes of early-onset severe preeclampsia.

Adult ; Asian Continental Ancestry Group ; genetics ; China ; Female ; Gestational Age ; Humans ; Infant, Newborn ; Male ; Polymorphism, Single Nucleotide ; Pre-Eclampsia ; enzymology ; genetics ; physiopathology ; Pregnancy ; Pregnancy Outcome ; Serine Endopeptidases ; genetics