Inherited thrombophilias and pre-eclampsia in Brazilian women

OBJECTIVE: The aim of the present study was to compare the distribution of G1691A, G20210A and C677T mutations in pre-eclamptic Brazilian women and in matched control women with an uncomplicated normal pregnancy. STUDY DESIGN: these mutations were investigated by PCR-RFLP in 83 normal pregnancies (control group) and in 30 pre-eclamptic pregnant women (severe form). RESULTS: G1691A mutation was detected neither in the control group nor in pre-eclamsia women. G20210A mutation was detected in heterozygosis in 3 (3.61%) control subjects, but not in pre-eclampsia group. C677T mutation was detected in homozygosis in 6 (7.23%) control subjects and 2 (6.67%) pre-eclamptic women and in heterozygosis in 31 (37.3%) control subjects and 12 (40%) pre-eclamptic women. Differences in the mutation frequencies detected in the two groups were not statistically significant. CONCLUSION: No correlation was observed between pre-eclampsia and presence of G1691A, G20210A and C677T mutations in Brazilian women.     

血清补体C1q肿瘤坏死因子相关蛋白3、亲环素A与子痫前期患者脂代谢异常的相关性研究

目的探讨血清补体C1q肿瘤坏死因子相关蛋白(CTRP)-3、亲环素A(CyPA)与子痫前期(PE)患者脂代谢异常的相关性。 方法选取2017年1月至2018年6月,青海大学附属医院产科收治的98例PE患者为研究对象。根据PE严重程度,将其分为重度PE组(n＝48)与轻度PE组(n＝50)。同时采用随机数字表法,选取同期于本院进行定期产前检查的50例健康孕妇作为对照,纳入对照组。采用酶联免疫吸附法,检测3组受试者的血清CTRP-3、CyPA水平。采用全自动生化分析仪,检测3组受试者的血清总胆固醇(TC)、甘油三酯(TG)、高密度脂蛋白胆固醇(HDL-C)与低密度脂蛋白胆固醇(LDL-C)水平。采用单因素方差分析,对3组受试者的人体质量指数(BMI)、收缩压、舒张压及血清TG、TC、LDL-C、HDL-C、CTRP-3、CyPA水平进行总体比较;然后,再采用最小显著性差异(LSD)法,进一步两两比较。PE患者血清CTRP-3、CyPA水平与其血清TG、TC、LDL-C、HDL-C水平的相关性,采用Pearson相关性分析。PE患者血清CTRP-3、CyPA水平与其脂代谢指标的多因素分析,采用多重线性回归法。本研究遵循的程序符合2013年修订的《世界医学协会赫尔辛基宣言》要求。3组受试者年龄、孕龄分别比较,差异均无统计学意义(P>0.05)。 结果①3组受试者BMI、收缩压、舒张压及血清TG、TC、LDL-C、HDL-C、CTRP-3、CyPA水平分别总体比较,差异均有统计学意义(F＝5.138、6.742、7.012、7.436、6.538、6.715、7.213、27.659、7.704,P<0.05)。对其进一步进行两两比较的结果显示,重度PE组和轻度PE组受试者BMI、收缩压、舒张压及血清TG、TC、LDL-C、CyPA水平,均分别显著高于对照组,并且差异均有统计学意义(重度PE组vs对照组：LSD-t＝7.156、13.864、8.664、14.264、6.837、13.875、28.185,P<0.001;轻度PE组vs对照组：LSD-t＝5.875、11.673、7.148、10.643、4.184、11.684、23.686,P<0.001);而血清HDL-C、CTRP-3水平,则均分别显著低于对照组,并且差异亦均有统计学意义(重度PE组vs对照组：LSD-t＝12.864、44.378,P<0.001;轻度PE组vs对照组：LSD-t＝9.547、31.684,P<0.001)。重度PE组受试者的BMI、收缩压、舒张压及血清TG、TC、LDL-C、CyPA水平,均显著高于轻度PE组,并且差异均有统计学意义(LSD-t＝4.738、P<0.001,LSD-t＝9.814、P<0.001,LSD-t＝4.864、P<0.001,LSD-t＝2.326、P＝0.042,LSD-t＝2.563、P＝0.028,LSD-t＝2.713、P＝0.022,LSD-t＝6.145、P<0.001);而血清HDL-C、CTRP-3水平,则均显著低于轻度PE组,差异亦均有统计学意义(LSD-t＝6.258、16.386,P<0.001)。②PE患者的血清CTRP-3水平与其血清TG、TC、LDL-C水平,均分别呈负相关关系(r＝－0.546、－0.573、－0.601,P<0.001),而其血清CTRP-3水平与血清HDL-C水平,则呈正相关关系(r＝0.531,P<0.001)。PE患者的血清CyPA水平与其血清TG、TC、LDL-C水平,均分别呈正相关关系(r＝0.612、0.637、0.552,P<0.001),而其血清CyPA水平与血清HDL-C水平,则呈负相关关系(r＝－0.514,P<0.001)。③血清TG、TC、LDL-C、HDL-C水平,均分别为PE患者血清CTRP-3、CyPA水平的独立影响因素(血清CTRP-3水平：t＝4.427、P<0.001,t＝3.675、P<0.001,t＝2.328、P＝0.020,t＝4.576、P<0.001;血清CyPA水平：t＝2.788、P＝0.005,t＝3.332、P＝0.001,t＝2.429、P＝0.015,t＝2.140、P＝0.032)。 结论PE患者血清CTRP-3水平较正常值降低、CyPA水平较正常值升高。CTRP-3、CyPA水平与其脂代谢异常具有相关关系,可能参与PE的发生、发展过程。     

nephrin、血管内皮生长因子及其受体表达与先兆子痫大鼠蛋白尿的关系

目的 研究nephrin、血管内皮生长因子（VEGF）及其受体（VEGFR）在先兆子痫大鼠肾组织的表达与蛋白尿的关系。 方法 采用一氧化氮合酶抑制剂（L-NAME）制备大鼠先兆子痫模型（n=8）,分别与正常雌性组（n=6）、正常妊娠组（n=8）、未孕L-NAME对照组（n=6）大鼠比较动脉收缩压（SBP）、尿蛋白量（24 h）。各组大鼠肾组织分别行光镜和电镜检查。Western印迹法、实时定量PCR检测nephrin在各组大鼠肾脏局部的表达;免疫荧光法检测各组大鼠肾小球内wilms肿瘤蛋白WT1的表达。Western印迹法检测VEGF及VEGFR（Flt-1、Flk-1）在各组大鼠肾脏局部的表达。 结果 先兆子痫模型组大鼠nephrin蛋白的表达（0.0726±0.0074）显著低于正常雌性组（0.3795±0.0509）、正常妊娠组（0.2361±0.0437）及未孕L-NAME对照组大鼠（0.7265±0.0503）（均P < 0.01）;而nephrin mRNA在各组大鼠间差异无统计学意义。各组大鼠足细胞数目差异无统计学意义。先兆子痫大鼠组VEGF的表达（1.5429±0.0898）显著高于正常雌性组（1.1870±0.1160）、正常妊娠组（1.3741±0.1165）、未孕L-NAME对照组大鼠（1.0155±0.0742）（均P < 0.01）;先兆子痫组大鼠VEGFR（Flt-1、Flk-1）的表达均显著高于其他各对照组大鼠（均P < 0.05）。 结论 先兆子痫大鼠中,nephrin蛋白水平表达明显降低,肾脏局部VEGF-VEGFR表达显著增强,可能参与了先兆子痫蛋白尿的生成,其具体机制有待进一步研究。     

基因微阵列技术检测子痫前期相关胎盘组织基因表达变化的研究

目的:应用基因微阵列技术检测子痫前期(pre-eclampsia,PE)患者和正常妊娠胎盘组织差异表达基因,寻找PE相关发病基因,为研究子痫前期病因提供分子水平理论基础。方法:从正常妊娠和子痫前期胎盘组织中抽提mRNA,反转录后,探针标记cDNA。以混合的6例正常胎盘总RNA为对照,应用cDNA微阵列人类全基因表达谱分析芯片检测6例子痫前期患者胎盘基因表达的改变。结果:在芯片杂交过程中,有91个基因表达差异具有统计学意义,与正常对照相比,子痫前期样品中有19个基因表达下调,72个基因表达上调,这些基因涉及信号传导、运输、转录调控、细胞增殖和凋亡、脂代谢、钙离子结合以及免疫等方面的功能。结论:子痫前期发病涉及了多方面细胞功能基因表达的变化,应用cDNA表达谱芯片可快捷、高通量地筛选出子痫前期胎盘可能的致病基因,为研究子痫前期的病因提供新的理论基础。     

孕妇桡动脉血流图对预测子痫前期的临床观察

目的探讨孕妇桡动脉血流图的监测在预测子痫前期中的应用价值,以准确识别有高危风险的孕妇,早期预测子痫前期的发生,减少母婴伤害。方法对产前检查无高血压、糖尿病、心、肝、肾等并发症,孕周为16～36周的10 433例孕妇分为预测组(5 223例)和对照组(5 210例)。预测组应用桡动脉血流图进行监测,每4周重复1次,对预测阳性者每2周重复测定并给予干预措施;对照组直到发现子痫前期症状时才给予干预。随访两组患者至分娩结束,对两组患者子痫前期的发生率、发病程度及新生儿体质量进行比较。结果预测组中子痫前期及子痫的发病率分别为11.2%和0.11%,对照组中子痫前期及子痫的发病率分别为13.5%和0.29%(P<0.05);新生儿生长受限率在预测组和对照组中分别为5.11%和7.06%(χ2=15.38,P<0.01);对照组中子痫前期的病情程度明显较预测组重(P<0.05);妊娠期高血压疾病监测系统在不同孕周预测子痫前期的敏感度不同,预测子痫前期的阳性符合率随孕周增加而增加。结论应用妊娠期高血压疾病监测系统及动态桡动脉血流动力学指标的改变早期预测子痫前期,对预测阳性者给予临床指导,可降低其发病率并防止病情发展,改善孕产妇和围产儿的结局。     

子痫前期血清IFN-γ、IL-6的水平与患者病情变化的相关性研究

目的探讨子痫前期血清IFN-γ、IL-6的水平与患者病情变化的相关性。方法 Elisa法检测血清IFN-γ、IL-6在30例轻度子痫和28例重度子痫前期及35正常孕妇中的水平。结果轻、重度组子痫前期患者血清IFN-γ水平显著高于正常对照组(P<0.05),重度组血清IFN-γ高于轻度组,差异具有显著性(P<0.05);血清IL-6的水平随着患者病情加重而呈降低趋势,但轻、重度组与正常对照组比较无统计学差异(P>0.05).血清IFN-γ/IL-6比值与患者病情呈正相关(P<0.05)。结论 IL-6和IFN-γ在妊娠期高血压疾病的发生、发展中发挥重要作用。     

子痫前期及子痫期患者联合检测血小板膜糖蛋白及凝血酶原的临床意义

目的研究子痫前期及子痫期患者血管内皮损伤和血小板活化状态及凝血系统活跃程度。方法采用全血流式细胞术分别检测32例重度子痫前期／子痫患者、16例轻度子痫前期患者、22例孕晚期妇女剖宫产术前及剖宫产术后72h内血小板膜糖蛋白Ⅰbα（GPⅠbot）、Ⅱb（GPⅡb）水平变化,采用血凝仪同期检测血浆凝血酶原时间（PT）和活化部分凝血活酶时间（APTT）。结果剖宫产术前,与轻度子痫前期组、正常孕晚期组比较,重度子痫前期及子痫组GPⅠbα和PT明显降低（P〈0．01）,术后两者72h内恢复至正常孕晚期水平,与术前比较差异显著（P〈0．01）;各组间剖宫产术前或术后GPⅡb变化无显著性差异（P〉0．05）;正常孕晚期组、轻度子痫前期组及重度子痫前期子痫组APTT递次延长但无统计学差异（P〉0．05）;术后上述三组APTT递次降低,但无显著性差异（P〉0．05）。GPⅠbα与PT呈正相关（r=0．371,P〈0．01）。结论孕晚期子痫前期及子痫患者存在血管内皮细胞损伤及血小板活化状态,引起外源性凝血系统活跃,而内源性凝血系统由于肝脏产生的凝血因子相对减少而活跃性相对降低。     

Pre-eclampsia but not pregnancy-induced hypertension is a risk factor for diabetic nephropathy in type 1 diabetic women

Aims/hypothesis Our aim was to study whether pre-eclampsia and pregnancy-induced hypertension are predictors of diabetic nephropathy in type 1 diabetic women. Materials and methods A total of 203 type 1 diabetic women, who were pregnant between 1988 and 1996 and followed at the Department of Obstetrics and Gynaecology in Helsinki, were re-assessed after an average of 11 years within the nationwide, multi-centre Finnish Diabetic Nephropathy Study. Diabetic nephropathy was defined as microalbuminuria, macroalbuminuria or end-stage renal disease. Results Patients with prior pre-eclampsia had diabetic nephropathy more often than patients with a normotensive pregnancy (diabetic nephropathy vs normal albumin excretion rate: 41.9% vs 8.9%; p<0.001), whereas patients with a history of pregnancy-induced hypertension did not (10.3% vs 8.9%; p=0.81). CHD was more prevalent in patients with a history of pre-eclampsia than in patients with a normotensive pregnancy (12.2% vs. 2.2%; p=0.03). Pre-eclampsia (odds ratio [OR] 7.7, 95% CI 1.6-36.1; p=0.01) and HbA_1c (OR 2.0, 95% CI 1.1-3.8; p<0.05) were associated with incident diabetic nephropathy even when adjusted for follow-up time, BMI, smoking, diabetes duration and age. Conclusions/interpretation These data suggest that a history of pre-eclamptic pregnancy but not pregnancy-induced hypertension is associated with an elevated risk of diabetic nephropathy. Electronic supplementary material Supplementary material is available in the online version of this article at and is accessible to authorised users.     

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...