ANP promotes HTR-8/SVneo cell invasion by upregulating protein kinase N 3 via autophagy inhibition.

Preeclampsia is a gestational disease characterized by two major pathological changes-shallow trophoblast invasion and impaired spiral artery remodeling. Atrial natriuretic peptide (ANP) is a kind of peptide hormone that regulates blood pressure, while the lack of active ANP participates in preeclampsia pathogenesis. However, the underlying mechanism of how ANP modulates trophoblasts function remains unclarified. Here, we performed isobaric tags for relative and absolute quantification (iTRAQ) in ANP-treated HTR-8/SVneo cells and identified Protein Kinase 3 (PKN3) as the downstream factor of ANP, which was downregulated in preeclamptic placenta. Chromatin immunoprecipitation analysis and luciferase assays showed that NFYA was one of the transcription factors for the PKN3 promoter, which was also regulated by ANP treatment in HTR-8/SVneo cells. Transmission electron microscopy and Western Blotting in HTR-8/SVneo cells indicated that ANP inhibited autophagy via AMPK-mTORC1 signaling, while excess autophagy was observed in preeclamptic placenta. The increased expression of PKN3 and enhanced cell invasion ability in HTR-8/SVneo cells induced by ANP could be abolished by autophagy activation or transfection with PKN3 shRNA or NFYA shRNA or NPR-A shRNA via regulating the invasion-related genes and the epithelial mesenchymal transition molecules. Our results demonstrated that ANP could enhance trophoblast invasion by upregulating PKN3 via NFYA promotion through autophagy inhibition in an AMPK/mTORC1 signaling-dependent manner.

Gestational weight gain targets during the second and third trimesters of pregnancy for women with gestational diabetes mellitus in China.

BACKGROUND/OBJECTIVES: Gestational weight gain (GWG) recommendations for pregnant women with gestational diabetes mellitus (GDM) in China are lacking. The present study aims to examine whether specific GWG targets for women with GDM can improve pregnancy outcomes in comparison with GWG according to the Institute of Medicine (IOM) targets. SUBJECTS/METHODS: Pregnant women diagnosed with GDM were selected from a retrospective cohort study of 8299 singleton pregnant women aged 18-45 years in 2012 (n = 1820). GWG ranges were calculated using a receiver operating characteristic (ROC) curve analysis (ROC targets) and the interquartile range (IR) method (the range from the 25th to 75th percentiles of the GWG among GDM women without adverse pregnancy outcomes, IR targets). RESULTS: The incidences of small for gestational age (SGA) births and pregnancy hypertension among women with GDM who gained weight within the ROC targets were lower than the incidences in women who gained weight within the IOM targets (SGA, 7.5% vs. 8.6%; pregnancy hypertension, 12.6% vs. 14.1%; both P < 0.05). GWG was associated with a risk of adverse pregnancy outcomes in the total sample (estimated values ranged from -2.95 to 2.08, all P < 0.05). No statistically significant associations between GWG and adverse pregnancy outcomes were observed in subgroups of pregnant women with appropriate GWGs according to the ROC, IR, and IOM targets. The ROC targets exhibited higher negative predictive values for adverse pregnancy outcomes than the IR and IOM targets. CONCLUSION: The ROC targets improved pregnancy outcomes and thus represent potential special GWG guidelines for women with GDM in China.

Mirror syndrome associated with heart failure in a pregnant woman: a case report.

Mirror syndrome is also known as Ballantyne syndrome, maternal hydrops, triple edema and pseudotoxemia. The disease can be difficult to diagnose, is related to pregnancy and can grievously endanger the health of both the mother and fetus. The pathogenesis of the disease has not been fully elucidated, and this disease may be confused with preeclampsia, even though distinguishing features can be identified. In this report case, we describe a pregnant woman who suffered from mirror syndrome associated with heart failure. After delivery, the heart failure symptoms also disappeared.

[Effects of human leukocyte antigen-G on p38 mitogen-activated protein kinase signaling pathway in HTR-8/SVneo cell line].

OBJECTIVE: To investigate the role of human leukocyte antigen-G (HLA-G) on the invasion and the molecular mechanism involved in this cellular progress in HTR-8/SVneo cell line. METHODS: There were three groups: groups of transfection, negative control and blank control, which corresponding to treatment by HLA-G specific siRNA, negative siRNA and only lipofectamine 2000 using lipofection technology in HTR-8/SVneo cell line. The efficiency of down-regulated of HLA-G was detected by reverse transcription-polymerase chain reaction and western blot analysis in mRNA and protein level, respectively. Changes of p38 mitogen-activated protein kinases (p-p38MAPK)/p38MAPK protein levels and the cell invasion were respectively detected by western blot analysis and transwell test. RESULTS: (1) The mRNA levels of HLA-G transfection group, negative control group and blank control group were 0.26±0.08, 0.71±0.11, 0.79±0.07, respectively. There was significant difference between transfection group and negative control group (P<0.01), while there was no significant difference between negative control group and blank control group (P>0.05). The efficiencies of down-regulated of HLA-G were (69.8±6.3)%, (14.9±2.2)%, 0 in transfection group, negative control group and blank control group respectively in mRNA level. (2) In protein levels, HLA-G were 0.20±0.15, 0.75±0.12, 0.76±0.21 in transfection group, negative control group and blank control group, respectively. There was significant difference between transfection group and negative control group (P<0.01), whereas there was no significant difference between negative control group and blank control group (P>0.05). The efficiencies of down-regulated of HLA-G were (81.1±14.4)%, (18.0±7.7)%, 0 in transfection group, negative control group and blank control group respectively. (3) The invasive number of transfection group, negative control group and blank control group were 57±38, 364±79 and 260±84, respectively, with a significant difference between transfection group and negative control group (P<0.01). There was no significant difference between negative control group and blank control group (P>0.05). (4) The p-p38MAPK/p38MAPK values of the HLA-G transfection group, negative control group and blank control group were 0.74±0.04, 0.47±0.09 and 0.36±0.21, respectively. HLA-G transfection group was significantly different compared with the other two groups (P<0.01). (5) Without or with SB203580, the p-p38MAPK/p38MAPK values of the HLA-G transfection group were 0.89±0.09 and 0.16±0.04, the values of negative control group and blank control group were 0.76±0.08, 0.14±0.03 and 0.51±0.05, 0.03±0.01, respectively. There was significant difference between without SB203580 and with SB203580 (P<0.01). (6) Without or with SB203580, the invasive number of transfection group were 51±13 and 90±21, respectively, which was significantly different (P<0.01). The invasive number of negative control group and blank control group were 290±52, 298±33 and 290±73, 264±64, respectively, which was no significant difference between without SB203580 and with SB203580 (P>0.05). CONCLUSIONS: HLA-G gene may regulate invasion of trophoblast-derived cell line HTR-8/SVneo via p38MAPK signaling pathway. The lower expression of HLA-G in trophoblast cells may lead to the occurrence of pathologic pregnancy.

[Effects on human leukocyte antigen-G on proliferation and invasion of JEG-3 cell line].

OBJECTIVE: To investigate the effect of human leukocyte antigen-G (HLA-G) on the growth and invasion of JEG-3 cell line and the role of HLA-G in the onset and development of pre-eclampsia. METHODS: The experiment was composed of three groups: groups of transfection, negative control and blank control, which corresponded to groups of HLA-G siRNA transfection, negative siRNA transfection and no transfection. HLA-G overexpressed choriocarcinoma cell line JEG-3 was used. The role of HLA-G in JEG-3 cell monolayer was examined by RNA interference technology using HLA-G specific small interfering RNA (siRNA). Expression of HLA-G was detected by reverse transcriptase-polymerase chain reaction and western blot analysis. Changes of cell cycle, apoptosis, proliferation and invasion were respectively detected by methyl thiazolyl tetrazolium (MTT), flow cytometry assay and transwell test. RESULTS: (1) The mRNA and protein levels of HLA-G control group and blank control group were 0.0013 +/- 0.0014, 0.0163 +/- 0.0007 and 0.1923 +/- 0.0384, 0.2184 +/- 0.0153, respectively, which were both significantly different (P<0.05); the number of negative transfection group was 0.1606 +/- 0.0133 and 0.2020 +/- 0.0132, which had no significant difference compared with blank control group (P>0.05). (2) The integral absorbance (IA) values of the HLA-G transfection group and blank control group were 0.44 +/- 0.04 and 0.75 +/- 0.13 respectively, which was significantly different (P<0.01); the IA value of negative control group was 0.69 +/- 0.10, which was not significantly different compared with blank group (P>0.05). (3) The ratios of G2/M and S phase cells in transfection group were (10.9 +/- 2.2)% and (58.6 +/- 0.8)% respectively, significantly different compared with the blank control group [(15.4 +/- 1.9)% and (52.9 +/- 2.3)% respectively; P<0.01]. (4) The ratio of early apoptosis cells in transfection group [(14.5 +/- 2.7)%] was significantly increased compared with negative [(5.3 +/- 1.1)%] and blank control group [(4.7 +/- 0.6)% ; P<0.01]. (5) The invasion number of transfection group and blank control group were 121+/- 12 and 452 +/- 17 respectively, with a significant difference between them (P<0.01). CONCLUSION: HLA-G is probably involved in the onset of preeclampsia by regulating proliferation and invasion of trophoblast.