Placental Dysferlin Expression is Reduced in Severe Preeclampsia

Dysferlin (DYSF) and myoferlin (MYOF), members of the ferlin family of membrane proteins, are co-expressed in human placental syncytiotrophoblast (STB). Although the role of these ferlin proteins in the placenta has yet to be established, it has been suggested that DYSF and MYOF may contribute to the stability of the apical STB plasma membrane. The release of STB-derived cellular debris increases in the setting of preeclampsia (PE), suggesting relative destabilization of the hemochorial interface. To test whether PE was associated with alterations in placental expression of DYSF and/or MYOF, a cross-sectional study was performed using specimens of villous placenta collected form women with severe PE (n = 10) and normotensive controls (n = 10). DYSF and MYOF expression were examined using quantitative real–time RT-PCR, immunoblotting, and immunofluorescence labeling of tissue specimens. Placental DYSF expression was 57% lower at the mRNA level (p = 0.03) and 38% lower at the protein level (p = 0.026) in severe PE as compared to normotensive subjects. There were no differences in placental MYOF protein or mRNA expression between these groups. No appreciable changes in the distribution of DYSF or MYOF within placental villi was observed in PE relative to control specimens. We conclude that DYSF expression is reduced in severe PE relative to gestational age-matched controls. As DYSF has a role in membrane repair, these data suggest a role for DYSF in the stability of the apical STB plasma membrane and may account, at least in part, for the increased shedding of microparticles from this membrane in PE.     

Differential Placental Gene Expression in Severe Preeclampsia

We investigated the global placental gene expression profile in severe preeclampsia. Twenty-one women were randomly selected from 50 participants with uncomplicated pregnancies to match 21 patients with severe preeclampsia. A 30 K Human Genome Survey Microarray v.2.0 (Applied Biosystems) was used to evaluate the gene expression profile. After RNA isolation, five preeclamptic placentas were excluded due to poor RNA quality. The series composed of 37 hybridizations in a one-channel detection system of chemiluminescence emitted by the microarrays. An empirical Bayes analysis was applied to find differentially expressed genes. In preeclamptic placentas 213 genes were significantly (fold-change ≥ 2 and p ≤ 0.01) up-regulated and 82 were down-regulated, compared with normal placentas. Leptin (40 fold), laeverin (10 fold), different isoforms of β-hCG (3–6 fold), endoglin (4 fold), FLT1 (3 fold) and FLT4 (2 fold) were up-regulated. PDGFD was down-regulated (2 fold). Several differentially expressed genes were associated with Alzheimer disease, angiogenesis, Notch-, TGFβ- and VEGF-signalling pathways. Sixteen genes best discriminated preeclamptic from normal placentas. Comparison between early- (<34 weeks) and late-onset preeclampsia showed 168 differentially expressed genes with oxidative stress, inflammation, and endothelin signalling pathways mainly involved in early-onset disease. Validation of the microarray results was performed by RT-PCR, quantitative urine hCG measurement and placental histopathologic examination. In summary, placental gene expression is altered in preeclampsia and we provide a comprehensive list of the differentially expressed genes. Placental gene expression is different between early- and late-onset preeclampsia, suggesting differences in pathophysiology.     

早孕期人绒毛趋化因子CCL2及其受体CCR2的表达

目的:分析人早孕绒毛组织趋化因子CCL2及其受体CCR2的转录和翻译表达。方法:收集人早孕期胎盘组织30例,分离出绒毛组织,采用半定量RT-PCR、免疫组织化学法分析正常人早孕绒毛组织CCL2/CCR2的表达。结果:人早孕绒毛滋养细胞中等水平转录和翻译CCL2及CCR2。结论:CCL2/CCR2表达于母-胎界面的胎儿面绒毛组织,可能参与滋养细胞功能调控。     

缺氧对重度子痫前期患者胎盘绒毛组织代谢足迹变化的影响

目的:探讨在不同氧浓度下体外培养正常妊娠孕妇及重度子痫前期患者胎盘绒毛组织培养液中代谢足迹的变化。方法:选择正常妊娠组(27例)及重度子痫前期组(27例)孕妇,在胎盘边缘、中间、近脐带处取绒毛组织,分别在1%氧浓度下及6%氧浓度下培养96小时,采用高效液相质谱色谱仪检测培养液中代谢产物,观察不同氧浓度下培养的绒毛组织代谢足迹的变化。结果:两组分别取自1%氧浓度和6%氧浓度下培养的同一胎盘边缘、中间、近脐带3点处胎盘绒毛组织代谢足迹的比较差异无统计学意义(P>0.05);1%氧浓度与6%氧浓度下培养的正常妊娠组胎盘绒毛组织代谢足迹比较,发现545个代谢物差异有统计学意义(P<0.05);1%氧浓度与6%氧浓度下培养的重度子痫前期组胎盘绒毛组织代谢足迹的比较差异无统计学意义(P>0.05),1%氧浓度下培养的正常妊娠组与6%氧浓度下培养的重度子痫前期组胎盘绒毛组织代谢足迹的比较差异无统计学意义(P>0.05)。结论:缺氧可能影响胎盘绒毛组织的代谢,在重度子痫前期的发病机制中起重要作用,而利用高效液相质谱色谱仪的代谢组学技术能测定在不同氧浓度下培养的正常妊娠组和重度子痫前期孕妇胎盘绒毛组织代谢足迹的变化,表明该方法适合于重度子痫前期的代谢组学研究。     

可溶性酪氨酸激酶受体1/胎盘生长因子比值在子痫前期诊断中的价值

目的:探讨血清、尿液可溶性酪氨酸激酶受体1/胎盘生长因子(sFlt-1/PLGF)比值在子痫前期的诊断价值.方法:检测及比较分析40例正常孕妇、38例轻度子痫前期与32例重度子痫前期孕妇血清及尿液中PLGF、sFlt-1及sFlt-1/PLGF比值的变化,将尿液PLGF、sFlt-1分别与血清PLGF、sFlt-1进行Person相关分析,并用ROC曲线分析血清、尿液中sFlt-1/PLGF比值对子痫前期的诊断价值.结果:轻度子痫前期组血清、尿液sFlt-1/PLGF比值高于正常对照组,重度子痫前期组sFlt-1/PLGF比值高于轻度子痫前期组,各组之间的差异均有统计学意义(均P＜0.01);尿液PLGF、sFlt-1分别与血清PLGF、sFlt-1具有正相关性(r=0.692,r=0.784,均P＜0.05);血清、尿液sFlt-1/PLGF比值对轻度子痫前期的ROC曲线下面积(AUC)分别为0.921、0.935,血清、尿液sFlt-1/PLGF比值对重度子痫前期的ROC的AUC分别为0.959、0.972.血清中sFlt-1/PLGF比值的AUC与尿液中sFlt-1/PLGF比值的AUC相比差异均无统计学意义(均P ＞0.05).结论:血清、尿液sFlt-1/PLGF比值在子痫前期患者中均有明显的升高,其为子痫前期的诊断提供了一个较理想的实验室指标,在临床中可以用尿液sFlt-1/PLGF比值替代血液sFlt-1/PLGF比值.     

妊娠高血压综合征患者胎盘绒毛组织内皮素1和内皮型一氧化氮合成酶的基因表达及其脐动脉血流变化

目的　研究妊娠高血压综合征 (简称妊高征 )患者胎盘绒毛组织内皮素 1(endothelin- 1,ET- 1)及内皮型一氧化氮合成酶 (endothelial nitric oxide synthase,e NOS)的基因表达 ,探讨 ET- 1及一氧化氮在妊高征患者胎儿胎盘循环中对脐动脉血流变化的作用。　方法　对妊高征患者及正常妊娠孕妇的胎盘绒毛组织 ,用地高辛标记的 ET- 1及 e NOS互补脱氧核糖核酸 (com plem entary deoxyri-bonucleotide,c DNA)探针进行点印迹杂交 ,利用 L eica QWIN图象处理系统测量每个杂交点的平均光密度值。在分娩前一日用彩色超声多普勒仪测定其脐动脉血流 S/ D值。　结果　妊高征患者胎盘绒毛组织 ET- 1基因表达明显增多 ,其平均光密度值妊高征组为 0 .43± 0 .0 3,正常组为 0 .2 3± 0 .44两组间差别显著 P<0 .0 5。而妊高征组胎盘绒毛组织 e NOS基因的表达明显低于正常对照组 ,其平均光密度值妊高征组为 0 .19± 0 .2 2 ,正常组为 0 .38± 0 .0 2 ,两组间差别显著 P<0 .0 5。妊高征组脐动脉S/ D值明显高于正常对照组 ,分别为 5 .92± 2 .13和 2 .11± 0 .2 2 ,两组间差别显著 P<0 .0 5。　结论　妊高征患者胎盘绒毛组织 ET- 1的表达明显增加 ,而内 e NOS的表达明显减少 ,脐动脉血流阻力亦明显增加。妊高征患者脐动脉血流     

子痫前期孕妇血清、胎盘组织MMP-9及NGAL表达变化及其意义的研究

目的:探讨孕妇血清基质金属蛋白酶-9(MMP-9)、中性粒细胞明胶酶相关脂质运载蛋白(NGAL)水平变化及胎盘组织中MMP-9mRNA、NGAL mRNA表达变化在子痫前期发病中的作用.方法:选取60例重度子痫前期孕妇,按发病时孕周不同分为早发型组(发病孕周≤34周)和晚发型组(发病孕周＞34周)各30例;另选30例正...     

Placental Microparticles,DNA, and RNA in Preeclampsia

Preeclampsia is a common disorder of the second half of pregnancy that complicates 2% to 7% of all pregnancies worldwide and remains a major cause of maternal and fetal morbidity and mortality. Although the origin of the disease is still elusive, population-based studies have suggested that it might implicate genetic, immunologic, or physiologic factors. On the other hand, there is no doubt that the placenta plays an important role in its development. In preeclampsia, the shedding of placenta debris, such as syncytiotrophoblast microparticles (STBMs) and DNA and messenger RNA molecules, into the maternal peripheral blood is increased. The analysis of this material may give new insight into placentation and the underlying etiology of this disorder, as well as yield new tracks of research for the understanding of the molecular mechanisms, leading to the generation of the clinical symptoms.     

子痫前期患者外周血及胎盘组织中半乳糖凝集素1的表达及意义

目的：探讨子痫前期患者外周血及胎盘组织中半乳糖凝集素1（galectin-1,Gal-1）的表达及意义。方法：选取我院收治的早发型重度子痫前期患者53例为观察组,同期入院体检正常的孕妇53例为对照组。采用酶联免疫法检测血清Gal-1的浓度,经免疫组织化学法检测胎盘组织Gal-1蛋白的表达。结果：观察组和对照组外周血Gal-1浓度分别为（38.59±9.02）ng/L和（27.33±8.31）ng/L,2组比较差异有统计学意义（t=6.684,P＜0.001）。观察组胎盘组织Gal-1的表达高于对照组,差异有统计学意义（Z=5.632,P＜0.001）。结论：Gal-1可能与早发型重度子痫前期的发病有关,或可成为有价值的预测子痫前期的生物标记物。     

子痫前期患者胎盘组织中降钙素受体刺激肽-3(CRSP-3)的表达及意义

目的:研究子痫前期患者胎盘组织中降钙素受体刺激肽-3(CRSP-3)的表达与正常妊娠者之间的差异及其临床意义。方法:采用免疫组织化学方法检测住院分娩的50例子痫前期患者(子痫前期组:轻度子痫前期25例,重度子痫前期25例)和30例正常晚期妊娠孕妇(对照组)胎盘组织中CRSP-3的表达定位,Real-time PCR及Western blotting方法检测胎盘组织中CRSP-3mRNA和蛋白的表达。结果:子痫前期和正常妊娠者胎盘组织细胞滋养细胞和合体滋养细胞中均表达CRSP-3。子痫前期患者胎盘组织中CRSP-3mRNA及其蛋白表达水平均高于正常妊娠组(P均0.05)。其中重度子痫前期患者胎盘组织中CRSP-3mRNA和蛋白表达水平显著高于正常对照组(P均0.01),虽重度子痫前期胎盘组织中CRSP-3表达略高于轻度子痫前期,但组间无统计学差异(P均0.05)。结论:CRSP-3上调可能参与子痫前期的发病,但是与疾病的进展及严重程度无关。     

Comparison of Serum Levels and the Placental Expression of Resistin Between Patients with Preeclampsia and Normal Pregnant Women

Objective. The aim of this study was to evaluate serum resistin levels in women with preeclampsia compared to those in normal pregnant women and normal non-pregnant women, and to examine placental resistin expression. Methods. Serum resistin levels were measured by enzyme-linked immunosorbent assay and placental resistin expression was determined by immunohistochemistry. Results. Serum resistin levels were significantly elevated in women with preeclampsia compared to normal pregnant women and non-pregnant women. There was no significant difference in placental resistin expression. Conclusion. The placenta may not be the origin of the resistin that contributes to increased serum levels in women with preeclampsia.     

Placental Mitochondrial DNA and Respiratory Chain Enzymes in the Etiology of Preeclampsia

Objective: To evaluate the occurrence of the most common mutations and deletions in mitochondrial DNA and deficiencies in the enzyme complexes of the mitochondrial respiratory chain in placentas from preeclamptic women.Methods: Mitochondria were isolated from the placentas of 17 preeclamptic or 25 control women, and the activities of mitochondrial respiratory chain complexes were measured. Deletions and three common point mutations of mitochondrial DNA were searched for by the Southern blot and polymerase chain reaction (PCR) methods from the same placentas.Results: Mean (± standard deviation) mitochondrial respiratory chain enzyme complex activities in placentas on protein basis (nmol/min/mg of protein) were similar in preeclamptics and controls (nicotinamide adenine dinucleotide, reduced form–ubiquinone oxidoreductase 25.84 ± 9.29 versus 31.02 ± 7.52; nicotinamide adenine dinucleotide, reduced form–cytochrome-c oxidoreductase 77.88 ± 42.24 versus 104.06 ± 56.73; succinate–cytochrome-c oxidoreductase 57.90 ± 13.83 versus 64.44 ± 20.16; cytochrome-c oxidase 106.43 ± 35.46 versus 128.37 ± 48.64, respectively) and they were similar also when referenced to the mitochondrial marker enzyme citrate synthase. The sample sizes in both patient and control groups were found to be large enough by post hoc test. Large-scale deletions or the common 5-kb and 7.4-kb deletions were not detected, even at the sensitivity level of PCR. The three most common point mutations were not found in either control or preeclamptic placental samples.Conclusion: Common mitochondrial DNA mutations seem to play no major role in the universal etiology of preeclampsia, as assessed by analysis of the mitochondrial genome and respiratory chain enzyme activities in vitro. This does not exclude possible alterations in the energy state of the preeclamptic placenta.     

胎盘法尼醇X受体的表达及其与妊娠期肝内胆汁淤积症关系的研究

目的:探讨法尼醇X受体(FXR)在正常晚孕胎盘和妊娠期肝内胆汁淤积症(ICP)胎盘的表达,及其与母血、脐血总胆汁酸(TBA)水平的关系,分析胎盘FXR在ICP病理机制中的作用.方法:收集ICP患者(ICP组)及正常晚孕妇女(对照组)胎盘组织及母血、脐血各33例,并将ICP患者根据母血血清TBA是否≥40 μmoL/L将ICP组分为轻度ICP组和重度ICP组.并测定母血、脐血TBA水平和胎盘组织中FXR mRNA的相对表达量.结果:①羊水胎粪污染的发生率:ICP组羊水污染发生率高于对照组(χ~2:7.543,P=0.013);重度ICP组高于轻度ICP组(χ~2=7.637,P=0.013);②胎盘组织中FXR mRNA的表达情况:ICP组胎盘FXR mRNA表达量高于对照组(z=-2.391,P=0.017);重度ICP组胎盘FXR mRNA表达量高于轻度ICP组(z=-2.391,P=0.017);③ICP组胎盘FXR mRNA表达量与母血、脐血TBA呈正相关(r_s=0.348,P=0.047;r_s=0.284,P=0.027);对照组胎盘FXR mRNA表达量与母血、脐静脉血TBA无相关性(r_s=-0.068,P=0.716;r_s=0.010,P=0.959).结论:ICP时增高的胆汁酸水平上调胎盘FXRmRNA的表达,胎盘FXR表达增加可能为ICP时胎盘的一种对抗胎儿胆汁淤积的保护性机制.     

MTHFR C677T Polymorphism is Not Associated with Placental Abruption or Preeclampsia in Finnish Women

Objective: The aim of our study was to examine genetic variability in the gene encoding methylenetetrahydrofolate reductase (MTHFR) and individual susceptibility to the placental abruption or preeclampsia. Methods: 362 women (133 with preeclampsia, 117 with placental abruption, and 112 healthy controls) were genotyped for C677T polymorphism in the MTHFR gene. Results: Similar genotype distributions were observed in the frequencies of C/C homozygotes (58.6%, 64.1%, and 57.1% for the three groups, respectively) and mutant homozygotes T/T (9.0%, 5.1% and 5.4%). No significant differences were detected in T allele frequencies (25.2%, 20.5%, and 24.1% for the three groups, respectively). Conclusions: MTHFR C677T polymorphism does not have a major role in the development of preeclampsia or placental abruption in the Finnish population.     

Tolerance of Human Placental Tissue to Severe Hypoxia and Its Relevance for Dual Ex Vivo Perfusion

In the dual ex vivo perfusion of an isolated human placental cotyledon it takes on average 20–30 min to set up stable perfusion circuits for the maternal and fetal vascular compartments. In vivo placental tissue of all species maintains a highly active metabolism and it continues to puzzle investigators how this tissue can survive 30 min of ischemia with more or less complete anoxia following expulsion of the organ from the uterus and do so without severe damage.There seem to be parallels between “depressed metabolism” seen in the fetus and the immature neonate in the peripartum period and survival strategies described in mammals with increased tolerance of severe hypoxia like hibernators in the state of torpor or deep sea diving turtles. Increased tolerance of hypoxia in both is explained by “partial metabolic arrest” in the sense of a temporary suspension of Kleiber's rule. Furthermore the fetus can react to major changes in surrounding oxygen tension by decreasing or increasing the rate of specific basal metabolism, providing protection against severe hypoxia as well as oxidative stress.There is some evidence that adaptive mechanisms allowing increased tolerance of severe hypoxia in the fetus or immature neonate can also be found in placental tissue, of which at least the villous portion is of fetal origin.A better understanding of the molecular details of reprogramming of fetal and placental tissues in late pregnancy may be of clinical relevance for an improved risk assessment of the individual fetus during the critical transition from intrauterine life to the outside and for the development of potential prophylactic measures against severe ante- or intrapartum hypoxia. Responses of the tissue to reperfusion deserve intensive study, since they may provide a rational basis for preventive measures against reperfusion injury and related oxidative stress. Modification of the handling of placental tissue during postpartum ischemia, and adaptation of the artificial reperfusion, may lead to an improvement of the ex vivo perfusion technique.     

Maternal Serum Human Placental Growth Hormone (hPGH) at 11 to 13 Weeks of Gestation in Preeclampsia

Objective. Human placental growth hormone (hPGH) is produced by human placenta and plays a central role in the maternal metabolic adjustments to pregnancy. The objective of this study was to investigate the maternal serum concentration of hPGH at 11–13 weeks of gestation in pregnancies that subsequently developed preeclampsia (PE), and to examine the possible association with uterine artery pulsatility index (PI) and maternal serum pregnancy-associated plasma protein-A (PAPP-A). Methods. The maternal serum concentration of hPGH at 11–13 weeks was measured in a case–control study from 60 cases that developed PE and 120 unaffected controls. The measured hPGH concentration was converted into a multiple of the expected median (MoM) in unaffected pregnancies. Regression analysis was used to determine the significance of association between hPGH MoM with uterine artery PI MoM and PAPP-A MoM. Results. In the pregnancies that subsequently developed PE the median serum hPGH concentration was not significantly different from that in the unaffected group (0.92 versus 1.00 MoM), whereas uterine artery PI was increased (1.31 versus 1.01 MoM) and serum PAPP-A was decreased (0.76 versus 1.01 MoM). In the group that developed PE there was no significant association between serum hPGH MoM and gestational age at delivery, uterine artery PI MoM, or serum PAPP-A MoM. Conclusion. The finding that in the PE group serum hPGH level during the first trimester is normal suggests that it is unlikely that this hormone plays a role in the pathogenesis of PE.     

子痫前期胎盘组织差异表达基因ABCG_2的克隆和鉴定

目的:克隆和鉴定子痫前期胎盘组织差异表达基因,确定与子痫前期相关的基因。方法:应用荧光mRNA差异显示技术寻找并克隆子痫前期胎盘组织差异表达的基因,对其中的ATP结合盒转运子G_2(ATP binding cassette transporter G_2,ABCG_2)基因用半定量RT-PCR技术验证其表达。结果:ABCG_2基因在子痫前期胎盘组织中高表达,半定量RT-PCR结果与mRNA差异显示的结果相符合,ABCG_2基因在子痫前期胎盘组织中的表达高于正常胎盘组织,差异有显著性(P<0.05)。结论:荧光mRNA差异显示技术可用于筛选子痫前期胎盘组织差异表达基因,并应用此技术发现了ABCG_2基因在子痫前期患者胎盘组织表达水平增高。