子痫前期胎盘中生长阻滞和DNA损伤45α基因及p38丝裂原活化蛋白激酶的表达

目的 探讨子痫前期胎盘组织中生长阻滞和DNA损伤45α(growth arrest and DNA damage-inducible 45 alpha,Gadd45α)基因和p38丝裂原活化蛋白激酶(p38 mitogen-activated protein kinase,p38 MAPK)信号分子的表达,及其与血清中可溶性血管内皮生长因子受体-1(soluble vascular endothelial growth factor receptor-1,sFlt-1)和可溶性endoglin(soluble endoglin,sEng)的相关性分析.方法 选择2009年9月至2010年3月在重庆医科大学附属第一医院住院分娩的孕妇54例为研究对象,按病情分为子痫前期轻度组20例、子痫前期重度组16例,以足月择期剖宫产孕妇18例为对照组.采用免疫组织化学SP法检测Gadd45α及磷酸化p38 MAPK(phospho-p38 MAPK,p-p38 MAPK)蛋白在各组胎盘组织中的定位;实时荧光定量PCR检测各组胎盘组织中Gadd45α mRNA水平;Western印迹法检测各组胎盘组织中Gadd45α蛋白的表达水平、p38 MAPK及p-p38 MAPK的蛋白表达差异;双抗体夹心酶联免疫吸附法检测各组孕妇血清样本中sFlt-1及sEng的含量,并进行单因素方差及LSD-t检验分析.结果 (1)免疫组织化学检测Gadd45α与p-p38 MAPK蛋白均定位于胎盘滋养层细胞胞质及胞核、血管内皮细胞胞核及少量间质细胞胞核.(2)子痫前期重度及轻度组Gadd45α mRNA相对表达水平(3.33±0.13和2.10±0.11)较正常对照组(1.01±0.18)明显上调,且子痫前期重度组高于轻度组,两两比较差异均有统计学意义(P＜0.05).(3)Western 印迹法检测正常对照组、子痫前期轻度组及重度组患者胎盘组织中Gadd45α蛋白表达水平分别为0.22±0.11、0.65±0.15、1.34±0.17;p-p38 MAPK蛋白的表达水平分别为0.32±0.08、0.72±0.12、1.45±0.21,两两比较差异均有统计学意义(P＜0.05);p38 MAPK蛋白水平组间比较差异无统计学意义(P＞0.05).(4)子痫前期轻度组、重度组孕妇血清sFlt-1、sEng水平明显高于正常对照组,且子痫前期重度组高于轻度组,两两比较差异均有统计学意义(P＜0.05).(5)各组Gadd45α蛋白水平与孕妇血清中的sFlt-1、sEng含量呈正相关(r分别为0.88和0.87,P均＜0.05).结论 Gadd45α在子痫前期患者胎盘组织中的表达明显升高,其可能通过调控p38 MAPK信号转导通路,诱使循环中的sFlt-1、sEng释放增加,从而加重胎盘血管重铸障碍和抑制滋养细胞浸润,参与子痫前期的发病.

Abstract：

Objective To evaluate the expression of Gadd45α and p38 MAPK in placentas and the correlations of Gadd45α protein and serum soluble vascular endothelial growth factor receptor-1 (sFlt-1) and soluble endoglin (sEng) in preeclampsia(PE). Methods Fifty-four pregnant women who delivered from September 2009 to March 2010 in the First Affiliated Hospital of Chongqing Medical University were chosen as the subjects. They were classified into mild preeclampsia group (n=20),severe preeclampsia group (n=16) and the control group (normal pregnant women underwent elective cesarean sections at term without labor and perinatal complications, n=18). Western blot and immunohistochemistry were employed to determine the expression and localization of Gadd45α and p-p38 MAPK protein respectively. Gadd45α mRNA level was determined by quantitative real-time PCR. The levels of seum sFlt-1 and sEng were measured by enzyme-linked immunosorbent assay (ELISA). One-way ANOVA and LSD-t test were applied for statistical analysis. Results (1)Immunohistochemistry identified that the positive stained cells were mostly located in trophoblast cells in normotensive placentas, whereas in preeclamptic placentas Gadd45α protein and p-p38 MAPK protein were detected in trophoblast and endothelial cells, as well as a few stromal cells at increased levels.(2)The mRNA levels of Gadd45α was significantly elevated in mild and severe preeclampsia groups compared to the control group (2.10±0.11 and 3.33±0.13 vs 1.01±0.18, P＜0.05), and Gadd45α mRNA level in severe group was significantly higher than in mild group (P＜0.05).(3)The data of Western blot revealed that the Gadd45α protein levels in each group were 0.22±0.11, 0.65±0.15 and 1.34±0.17, respectively, with significant differences between each group(P＜0.05). The p-p38 MAPK protein levels in each group were 0.32±0.08, 0.72±0.12 and 1.45±0.21, respectively, with significant differences between each group (P＜0.05). p38 MAPK protein levels in the total groups showed no difference(P＞0.05).(4)Compared with the control group, sFlt-1 and sEng concentrations in maternal circulation were significantly increasing in mild and severe preeclampsia groups, and concentrations in severe group were significantly higher than those in mild group (P＜0.05).(5) There were positive correlations between Gadd45α protein levels and the concentrations of serum sFlt-1 and sEng in each group( r=0.88 and 0.87, respectively all P＜0.05). Conclusions Upregulation of Gadd45α in preeclampsia placentas may play an important role in the pathogenesis of preeclampsia. It may induce the increased maternal serum levels of sFlt-l and sEng by activating p38 MAPK signaling pathway, leading to deficient cytotrophoblastic invasion and abnormal placental vascular reconstruction during pregnancy.     

内脂素与子痫前期

内脂素(visfatin)是新近发现的一种主要由内脏脂肪细胞分泌的脂肪因子,结构复杂,存在基因多态性,具有调节糖脂代谢作用.内脂索增加胰岛素敏感性、促进血管平滑肌细胞成熟、参与炎症反应及血管生成,与动脉粥样硬化形成及血管内皮细胞受损和功能紊乱密切相关.子痫前期是一种病因不明的病理妊娠,严重危害母要生命,血管内皮细胞损伤...     

内脂素/前B细胞克隆增强因子与妊娠

内脂素/前B细胞克隆增强因子(visfatin/PBEF)作为新发现的脂肪因子,主要在内脏脂肪和大网膜中表达.具有类胰岛素作用、参与炎症反应和免疫调节等功能.妊娠期妇女处于特殊的生理状态,能量代谢发生特征性改变.最新观点认为妊娠合并症,如妊娠期糖尿病和妊娠期高血压疾病均存在胰岛素抵抗,是妊娠妇女体内代谢功能紊乱所致.近...     

Antiphospholipid antibodies internalised by human syncytiotrophoblast cause aberrant cell death and the release of necrotic trophoblast debris

Antiphospholipid antibodies (aPL) are the strongest maternal risk factor for pre-eclampsia, a hypertensive disease of human pregnancy. Pre-eclampsia is triggered by a toxic factor released from the placenta that activates the maternal endothelium. Antiphospholipid antibodies cause the release of necrotic trophoblast debris from the placental syncytiotrophoblast and this debris can activate endothelial cells. In this study, we investigated how aPL affects syncytiotrophoblast death and production of necrotic trophoblast debris by examining the interaction between aPL and human first trimester placental explants. Human polyclonal and murine monoclonal aPL, but not control antibodies, were rapidly internalised by the syncytiotrophoblast. Inhibitors of endocytosis or the low-density lipoprotein receptor (LDLR) family, but not toll-like receptors, decreased the internalisation of aPL and prevented the release of necrotic trophoblast debris from the syncytiotrophoblast. Once internalised, aPL increased inner mitochondrial membrane leak and Cytochrome c release while depressing oxidative flux through Complex IV of the electron transport system in syncytiotrophoblast mitochondria. These data suggest that the human syncytiotrophoblast internalises aPL by antigen-dependent endocytosis involving LDLR family members. Once internalised by the syncytiotrophoblast, aPL affects the death-regulating mitochondria, causing extrusion of necrotic trophoblast debris which can activate maternal endothelial cells thereby contributing to the pathogenesis of pre-eclampsia.     

Does contemporary ART lead to pre-eclampsia? A cohort study and meta-analysis

PurposeRecent publications suggested that the risk for pre-eclampsia (PE) is higher with frozen-thawed embryo transfers (FETs) compared to fresh transfers (IVF-ETs). These studies were based on old data that reflects outdated practices. In this paper, we wanted to assess the incidence of PE in current assisted reproductive technology (ART) practice.MethodsIn this cohort study, we present the incidence of PE in all births in the province of Ontario, Canada, for the years 2013–2017 for FET, IVF-ET, and natural conceptions (NC). We also compare our findings to previous studies in a meta-analysis that includes over 4 million births.ResultsThe results of our study show that contemporary practice of ART results in comparable risk for PE between FET and IVF-ET; however, the risk is higher than NC.ConclusionCurrent ART practice is associated with a lower risk for PE in frozen embryo transfer; this RR can be further attenuated by using ovulatory endometrial preparation for FETs.Supplementary InformationThe online version contains supplementary material available at 10.1007/s10815-021-02061-z.     

子痫前期患者脉压与蛋白尿相关性分析

目的：探讨子痫前期患者脉压与蛋白尿的相关性。方法：按脉压高低将子痫前期患者分为脉压≤40mmHg、41～80mmHg、〉80mmHg三组,比较各组蛋白尿的发生率。结果：三组间的收缩压、舒张压、蛋白尿发生率,24h尿蛋白浓度差异有统计学意义（P〈0.05）;SBP、DBP、平均动脉压和脉压与24h尿蛋白量的β值分别为0.425、0.331、0.357和0.486（P〈0.05）。结论：脉压对尿蛋白的发生率有明显影响,临床上在治疗子痫前期患者时,不但要降低收缩压和舒张压,还要降低脉压,只有这样才能使肾脏的损伤程度减轻。     

Managing lupus patients during pregnancy

Systemic lupus erythematosus (SLE) is an auto-immune disease, primarily affecting young females. Pregnancy in a woman with SLE remains a high-risk situation with higher maternal and foetal mortality and morbidity. Although live births are achieved in majority of the pregnancies, active disease and major organ involvement can negatively affect the outcomes. A higher risk of foetal loss, pre-term birth, intra-uterine growth restriction (IUGR) and neonatal lupus syndromes (NLSs) are major foetal issues. Mothers are faced with disease flares, pre-eclampsia and other complications. Disease flares during SLE pregnancy pose the unique issue of recognition and differentiation between physiologic changes and disease state. Similarly, pre-eclampsia and lupus nephritis may lead to diagnostic confusion. Treatment choices during pregnancy are limited to a few safe drugs, further restricting the options. Refractory pregnancy loss associated with anti-phospholipid antibodies (aPLs) and complete heart block associated with anti-Ro antibodies remain unresolved issues. A multidisciplinary approach, with close monitoring, is essential for optimal outcomes.     

妊娠期高血压疾病患者胎盘组织中血管活性肽Apelin的表达及其意义

目的 探讨妊娠期高血压疾病患者胎盘组织中血管活性肽Apelin的表达及其意义。方法 选择妊娠期高血压疾病患者36例（妊娠期高血压疾病组），其中妊娠期高血压10例，轻度子痫前期11例，重度子痫前期15例；选择同期正常产妇15例作为正常妊娠组。采用免疫组化染色法及实时荧光定量RT-PCR分别检测两组产妇胎盘组织中Apelin-36及ApelinmRNA的表达。结果 正常妊娠组Apelin-36及ApelinmRNA分别为0．27±0．04及0．82±0．25，妊娠期高血压疾病组分别为0．18±0．05及0．31±0．21，两组比较，差异有统计学意义（P〈0．01）；其中，妊娠期高血压患者分别为0．24±0．02及0．59±0．16，轻度子痫前期患者分别为0．16±0．03及0．25±0．07，重度子痫前期患者分别为0．14±0．02及0．17±0．09，轻度、重度子痫前期患者的Apelin-36及ApelinmRNA表达水平均低于妊娠期高血压患者（P〈0．01，P〈0．01），轻度子痫前期患者与重度子痫前期患者比较，差异也有统计学意义（P〈0．05，P〈0．05）。结论 血管活性肽Apelin在胎盘组织中表达降低，可能与妊娠期高血压疾病的发生、发展有关。     

同型半胱氨酸对滋养细胞基质金属蛋白酶2、9表达的影响

目的探讨同型半胱氨酸（Hcy）对早孕期滋养细胞基质金属蛋白酶（MMP）2、9表达的影响及其在子病前期发病过程中的作用。方法分离培养早孕期滋养细胞，用Hcy刺激早孕期滋养细胞48h，对照组Hcy刺激浓度为0mmol／L，实验组的刺激浓度为1mmol／L；以实时荧光定量RT-PCR检测早孕期滋养细胞MMP-2、MMP-9mRNA的表达变化；明胶酶谱分析法测定早孕期滋养细胞MMP-2、MMP-9酶活性变化，以积分吸光度值表示。结果实验组与对照组比较，早孕期滋养细胞MMP-2mRNA表达降低21％，MMP-9mRNA表达降低11％；实验组与对照组比较，MMP-2蛋白表达降低14％，MMP-9蛋白表达降低52％。结论Hey能够抑制早孕期滋养细胞MMP02、MMP-9的表达，从而影响滋养细胞侵袭性。