可溶性Flt-1在子痫前期患者胎盘组织中的变化及意义

目的研究可溶性血管内皮生长因子受体1(soluble fms-like tyrosine kinase receptor 1, sFlt-1)mRNA在子痫前期患者胎盘组织中的表达与定位,探讨其与子痫前期的关系.方法通过半定量RT-PCR及原位杂交方法分别检测子痫前期患者和健康孕妇胎盘组织中sFlt-1 mRNA的表达及定位.结果正常胎盘组织和子痫前期胎盘组织中均存在sFlt-1 mRNA,子痫前期胎盘组织中sFlt-1 mRNA表达明显高于正常胎盘组织(P<0.01).sFlt-1 mRNA主要分布在胎盘绒毛滋养细胞及血管内皮细胞胞浆内.结论胎盘组织中sFlt-1的高表达可能参与了子痫前期的病理生理过程.     

哈萨克族和汉族子痫前期患者的血脂变化与胎盘组织中脂蛋白酯酶表达的相关研究

目的探究哈萨克族和汉族子痫前期产妇的血脂变化与脂蛋白酯酶(LPL)在胎盘中表达的相关性。方法选取2016年9月至2017年3月在新疆医科大学第一附属医院产科确诊入住的100例子痫前期产妇为研究对象,依据民族不同分为各50例产妇的哈萨克族子痫前期组和汉族子痫前期组,另随机抽取同期汉族正常产妇50例作为对照组。采用酶法检测血脂浓度,ELISA法检测血清LPL浓度,RT-PCR和WesternBlot法检测胎盘组织LPL基因及蛋白表达水平。结果哈萨克族及汉族子痫前期组产妇血脂中总胆固醇(TC)、甘油三酯(TG)及低密度脂蛋白(LDL)浓度均高于对照组(P0.05),高密度脂蛋白(HDL)浓度低于对照组(P0.05),两子痫前期组产妇血脂中LDL、HDL及TG浓度比较存在明显差异(P0.05);两子痫前期组产妇血清LPL浓度均低于对照组(P0.05),哈萨克族子痫前期组产妇的血清LPL浓度低于汉族子痫前期组(P0.05);哈萨克族和汉族子痫前期组产妇胎盘组织LPL基因及蛋白的表达水平均低于对照组(P0.05);两子痫前期组产妇的血清LPL浓度与HDL(r=0.766、0.661)、胎盘组织LPL蛋白(r=0.613、0.427)呈显著正相关(P0.05),与TG呈显著负相关(r=-0.732、-0.594,P0.05)。结论哈萨克族和汉族子痫前期产妇血脂变化受多种因素调控,且与产后胎盘组织中LPL的表达具有相关性。     

NO及VEGF、sFlt-1在早发型子痫前期胎盘中的表达及意义

目的检测VEGF、sFlt-1及NO在早发型子痫前期和晚发型子痫前期患者血清和胎盘中的表达。方法随机选取2013年5月至2014年6月规律产检并分娩的子痫前期孕妇75例,其中早发型子痫前期31例,晚发型子痫前期44例。选取同期健康晚期妊娠孕妇30例作为健康对照组,采用免疫组织化学SP法检测VEGF、sFlt-1的表达,用硝酸还原法测定胎盘中NO的表达,并进行相关性分析。结果早发型子痫前期患者胎盘组织中VEGF阳性表达率32.2%,晚发型组为61.4%,正常对照组为86.7%,3组差异有统计学意义(P0.05);早发型子痫前期患者胎盘组织中sFlt-1阳性表达率82.1%,晚发型组74.8%,正常对照组12.9%,3组差异有统计学意义(P0.05);NO的光密度结果在早发型子痫前期患者胎盘组织中是59.3±2.21,晚发型组为31.6±1.90,正常对照组89.9±2.17,3组差异有统计学意义(P0.05)。结论 sFlt-1在早发型子痫患者的胎盘组织中表达最高,抑制了其靶组织VEGF表达,影响了胎盘微血管形成;NO在早发型子痫患者的胎盘组织中表达最低,抑制了微血管的舒张。SFlt-1和NO与早发型子痫前期的发病密切相关。     

KiSS-1在子痫前期患者胎盘组织中的表达及意义

目的探讨肿瘤转移抑制基因KiSS-1是否是子痫前期的发病机制。方法采用免疫组化SP法检测25例正常足月妊娠妇女(正常妊娠组)与40例子痫前期患者(子痫前期组,其中轻度子痫前期15例、重度子痫前期25例)胎盘组织中KiSS-1的表达。结果KiSS-1主要位于胎盘绒毛小叶的合体滋养细胞。子痫前期组胎盘组织中KiSS-1的平均光密度为(0.137±0.010),其中轻度子痫前期为(0.132±0.004),重度子痫前期为(0.140±0.012);均明显高于正常妊娠组的(0.124±0.010),P值分别为P<0.01、P<0.05(P=0.019)、P<0.01。结论胎盘组织中KiSS-1的表达水平随病情程度的加重而增高,可能与子痫前期的发病有关。     

MVD、VEGF、PLGF、sFlt-1在早发型子痫前期胎盘中的表达及意义

目的探讨VEGF、s Flt-1、PLGF、MVD在早发型子痫前期患者胎盘中的表达与胎盘血管病变的关系,并与晚发型子痫前期做比较。方法随机选取2013年5月至2014年6月规律产检并分娩的子痫前期孕妇105例,其中早发型子痫前期46例,晚发型子痫前期59例。选取同期健康孕妇50例作为健康对照组,采用免疫组织化学SP法检测MVD、VEGF、PLGF、s Flt-1的表达,CD34双标免疫组化法标记血管检测胎盘中的微血管密度值,并进行相关性分析。结果早发型子痫前期患者胎盘组织中VEGF阳性表达率30.4%,晚发型组为61.0%,正常对照组为90%,3组差异有统计学意义(P0.05);早发型子痫前期患者胎盘组织中s Flt-1阳性表达率80.4%,晚发型组76.3%,正常对照组14%,3组差异有统计学意义(P0.05);早发型子痫前期患者胎盘组织中PLGF阳性表达率21.7%,晚发型组为64.0%,正常对照组88%,3组差异有统计学意义(P0.05);早发型子痫前期患者胎盘组织中微血管密度(37.12±2.16),晚发型组(59.35±2.78),正常对照组(90.10±4.18),三组差异有统计学意义(P0.05)。结论 s Flt-1在早发型子痫患者的胎盘组织中高表达,抑制了其靶组织VEGF及PLGF表达,影响了胎盘微血管形成,引发了子痫前期的发生,s Flt-1出现越早,子痫前期发生越早。     

内皮型一氧化氮合酶运输介导物在子痫前期患者胎盘组织中的定位及定量研究

目的探讨内皮型一氧化氮合酶运输介导物(endothelial n itric oxide synthase traffic inducer,NOSTR IN)在子痫前期(pre-ec lampsia,PE)患者胎盘血管内皮细胞中表达的变化及其在子痫前期发病过程中的作用。方法HE染色后镜下观察胎盘组织及血管的病理变化,免疫组织化学方法及W estern b lot检测子痫前期患者胎盘组织中NOSTR IN的表达。结果HE染色显示子痫前期患者胎盘绒毛血管变细,数目减少,血管合体膜增厚,纤维素样坏死明显多于正常妊娠;免疫组织化学显示正常妊娠和子痫前期患者胎盘血管内皮细胞中都有NOSTR IN的表达,但子痫前期患者胎盘血管内皮细胞胞浆染色较正常妊娠明显增强;W estern b lot显示子痫前期患者胎盘组织中NOSTR IN的表达显著高于正常妊娠(P<0.01)。结论胎盘组织中NOSTR IN表达增加可能是子痫前期发病机制的重要环节之一。     

血清可溶性Fas与胎盘组织胎盘生长因子的表达在早发型重度子痫前期发病中的作用

目的:探讨血清可溶性Fas与胎盘组织胎盘生长因子在早发型重度子痫前期发病中的作用。方法:采用ELISA和免疫组化法分别检测20例早发重度子痫前期患者、35例晚发重度子痫前期患者和40例正常晚期妊娠患者血清可溶性Fas(Soluble Fas,sFas)和胎盘组织胎盘生长因子(placental growth factor,PlGF)的表达。结果:正常妊娠组、早发重度组、晚发重度组血清sFas水平分别为(1.17±0.53)、(6.68±0.97)和(3.17±0.74)mg/L,早发重度组、晚发重度组均显著高于正常对照组,差异有显著性(P<0.05)。早发重度组又明显高于晚发重度组,差异有显著性。正常妊娠组胎盘组织PlGF表达的平均灰度值显著高于早发型及晚发型重度子痫前期组,其中早发型子痫前期重度组表达最低。早发重度组中血清sFas与PlGF呈负相关,相关系数-0.748。结论:重度子痫前期患者血清sFas表达升高,且早发重度组表达显著高于晚发重度组,并与PlGF呈负相关,说明其异常表达可能是早发重度子痫前期发病的主要原因。     

IL-21、IL-22在子痫前期患者胎盘组织中的表达水平及意义

目的通过研究子痫前期患者与正常孕妇胎盘组织中白介素-21(IL-21)和白介素-22(IL-22)的水平,探讨IL-21和IL-22在子痫前期发生机制中的作用。方法采用酶联免疫吸附法(ELISA)分别测定轻度、重度子痫前期患者和正常孕妇胎盘组织中IL-21和IL-22的水平,比较3组间细胞因子的表达水平。结果通过比较发现,轻度、重度子痫前期组IL-21、IL-22在胎盘组织中的表达与对照组比较,差异有统计学意义(P0.05)。但是,虽然重度子痫前期组IL-21、IL-22在胎盘组织中的表达均高于轻度子痫前期组,但是差异无统计学意义(P0.05)。结论子痫前期组患者胎盘组织中IL-21、IL-22的表达增强可能与子痫前期的发生、发展有关。     

子痫前期患者胎盘组织中尾加压素Ⅱ及一氧化氮的表达及意义

目的研究尾加压素Ⅱ（urotensin Ⅱ,UⅡ）及一氧化氮在子痫前期患者胎盘组织的表达情况及其与子痫前期发病的关系。方法采用RT-PCR方法检测45例子痫前期患者（轻度22例,重度23例）及20例正常晚期妊娠孕妇胎盘组织中UIImRNA的表达情况;同时用硝酸还原酶法检测子痫前期组孕妇与正常组孕妇胎盘组织中NO浓度。结果重度子痫前期患者胎盘组织UIImRNA（0.85±0.40）明显高于正常妊娠组（0.38±0.30）（P〈0.05）;轻度子痫前期患者胎盘组织UIImRNA（0.64±0.31）,与正常妊娠组比较差异无统计学意义（P〉0.05）。轻、重度子痫前期患者胎盘组织NO浓度分别为114.42±6.52 u/mg、79.22±3.31 u/mg,均显著低于正常妊娠组241.36±13.24u/mg（P〈0.05）。结论子痫前期胎盘组织中尾加压素Ⅱ水平升高,NO浓度下降,可能在子痫前期的发病中起一定作用。     

TGFβ1和明胶酶B在子痫前期胎盘中的表达及相关性研究

目的通过检测子痫前期患者胎盘组织中TGFβ1和MMP-9(明胶酶B)的表达,探讨其在子痫前期发病机制中的作用及两者之间的相关性。方法用SP免疫组化染色法观察正常妊娠(15例),子痫前期(24例)剖宫产后胎盘组织中TGFβ1和MMP-9的分布与定位,通过HMIAS-2000型全自动医学彩色图像分析系统对其进行定量分析。结果在重度子痫前期组胎盘组织中TGFβ1表达明显高于正常对照组和轻度组(P<0.05),轻度组与正常组比较差异无显著性(P=0.087);MMP-9在轻、重子痫前期组胎盘组织中表达均明显低于正常对照组(P<0.05);胎盘组织中TGFβ1的表达水平和MMP-9的表达水平呈负相关(P<0.05)。结论胎盘组织中TGFβ1、MMP-9表达异常引起滋养细胞浸润不足,提示两者可能并共同参与子痫前期的发生和发展。     

Expression profile of trophoblast invasion-associated genes in the pre-eclamptic placenta

The primary pathology of pre-eclampsia is thought be a defect in placentation due to failure of trophoblast invasion. Here, we aim to identify the expression profile of invasion-associated genes in the pre-eclamptic placenta. Messenger RNA (mRNA) expression levels of extracellular matrix molecule-related genes in five pre-eclamptic placentas and in five strictly matched normal placentas were assayed using complementary DNA (cDNA) microarrays representing over 220 human cytokine-associated or hormone-associated genes. Results demonstrated greater than two-fold higher expression of 18 extracellular matrix molecule genes, including cadherin, collagen, integrin and selectin, in the pre-eclamptic placenta. Extracellular matrix molecule degradation-related genes, including matrix metalloproteinase (MMP)-10, MMP-13, MMP-15, tissue inhibitor of metalloproteinase (TIMP)-2, TIMP-3, plasminogen and plasminogen activator, were also highly expressed in the pre-eclamptic placenta, compared to the normal placenta. Results suggest that the abnormal expression profiles of extracellular matrix molecules and degrading proteinases might be associated with the pathogenesis of pre-eclampsia.     

The etiological role of allogeneic fetal rejection in pre-eclampsia

PROBLEM: It has been demonstrated that allogeneic fetal rejection in normal pregnancy is prevented by placental indoleamine 2,3-dioxygenase (IDO). Further, an immunological etiology has been implicated in pre-eclampsia. METHOD OF STUDY: We examined the differences in placental IDO activity between normal and pre-eclamptic pregnancies. RESULTS: IDO mRNA expression and enzyme activity levels in the placenta were low in patients with severe pre-eclampsia. The enzyme activity also inversely correlates with the blood pressure of the patients. In the placentas from severe pre-eclampsia, IDO immunoreactivity was low, whereas regional T-cell infiltration was observed reciprocally proportional to the IDO activity. CONCLUSION: Our findings implicate a potential role for IDO activity and a maternal immunological reaction against an allogeneic fetus in the etiology of pre-eclampsia.     

Immunolocalization of alphaV,alpha3 and beta1 integrins in the human placenta with pre-eclampsia

Signs of pre-eclampsia are considered to be caused by maternal endothelial dysfunction due to circulating factors of placental origin. Integrins are a large family of cell surface, proteins that serve as receptors involved in cell-cell and cell-matrix interactions during placentation. Therefore, low expression of integrins or the lack of it may be encountered during pre-eclampsia. In the present study, we investigated the immunolocalisation of integrins alphaV, alpha3 and beta1 in placentas of normal and pre-eclamptic women. Thirty-two placentas from pre-eclamptic (n = 14) and normotensive (n = 18) women were used. Immunohistochemical staining was performed on formalin-fixed, paraffin-embedded tissue specimens, using anti-alphaV, anti-alpha3 and anti-beta1 antibodies and the indirect immunoperoxidase technique. A semi-quantitative grading system (HSCORE) was used to compare immunohistochemical staining intensities. Distribution patterns of alphaV, alpha3 and beta1 integrins were detected in cytotrophoblasts and Hofbauer cells in normal and pre-eclamptic placentas. Immunostaining of alphaV and beta1 integrins was slightly decreased in pre-eclamptic samples but alpha3 integrin immunostaining was similar in pre-eclamptic and normal placentas. Decreased immunostaining of integrins in the cytotrophoblasts may considered to be a structural basis for decreased placental perfusion in pre-eclampsia.     

ERBB4/HER4在子痫前期胎盘中的表达

目的探讨子痫前期患者胎盘滋养细胞人表皮生长因子受体4(ERBB4/HER4)的表达及意义。方法采用RT-PCR法检测35例子痫前期患者(子痫前期组)胎盘ERBB4的表达及其与胎盘重量及新生儿出生体重的关系,并与20例正常妊娠妇女(正常妊娠组)比较。结果在子痫前期患者胎盘中,ERBB4 mRNA表达明显高于正常妊娠胎盘,两者有显著性差异(P<0.05)。结论子痫前期患者胎盘ERBB4表达显著增高,与子痫前期的发病密切相关。     

Increased cystatin C expression in the pre-eclamptic placenta

Trophoblast invasion is regulated by proteinases and their inhibitors. Cystatin C inhibits cysteine proteinases. The serum concentration of cystatin C is increased in late pregnancy and pre-eclampsia. We aimed to investigate whether the expression of cystatin C is increased in the pre-eclamptic placenta and to investigate the expression pattern of cystatin C mRNA and protein in placental tissue. Tissue samples from the central part of the placenta from 13 normal and 22 pre-eclamptic pregnancies were included. We used real-time polymerase chain reaction (RT-PCR) and in situ hybridization for mRNA expression analysis and immunohistochemistry and Western blotting for protein expression analysis. RT-PCR showed a significantly higher expression of cystatin C mRNA in pre-eclampsia than in normal pregnancy, with the highest expression in cases with severe pre-eclampsia. In situ hybridization revealed a distinct pattern of high expression in the extravillous trophoblast cells of the basal plate and low expression in the syncytiotrophoblast covering villi. The cystatin C protein distribution matched the mRNA expression pattern. Western blot analysis revealed an increased protein expression in cases with severe pre-eclampsia and confirmed the presence of cystatin C in amniotic fluid samples. The high expression of cystatin C mRNA in the extravillous trophoblast cells of the basal plate suggests a role for cystatin C in the regulation of proteases in placentation. Placental expression and secretion of cystatin C could contribute to the elevated maternal plasma levels seen in pre-eclampsia.     

Necrotic feature of the trophoblasts lacking HLA-G expression in normal and pre-eclamptic placentas

PROBLEM: Human leukocyte antigen (HLA)-G is thought to be expressed in all placental extravillous trophoblasts (EXTs). In pre-eclamptic placentas, a lack of HLA-G expression on EXTs had been found, and deduced as a possible cause of pre-eclampsia. However, a subset of EXTs lacking expression of HLA-G can also be found in normal placenta. Therefore, we sought to compare these cells in normal and pre-eclamptic placentas. METHODS OF STUDY: Frozen sections of normal and pre-eclamptic placentas were examined by immunohistochemical staining using HLA-G monoclonal antibody 87G, histochemical enzymatic analysis of succinate dehydrogenase (SDH) and ultrastructural analysis. RESULTS: A subset of EXTs lacking HLA-G expression was found in both normal and pre-eclamptic placentas. These cells showed necrotic features such as the swelling of cells, eosin-achromatophilia, the loss of SDH activity and swelling mitochondria. Cells from both tissues were identical with regard to these features. CONCLUSION: The features of the EXTs lacking HLA-G expression indicated they had undergone necrosis and thus could not express HLA-G protein. Therefore, an alternative interpretation to the lack of HLA-G expression in pre-eclamptic placentas is that it is the result of cell death and not the cause.     

Immunohistochemical adrenomedullin expression is decreased in the placenta from pregnancies with pre-eclampsia

The purpose of the present study was to investigate whether placental immunohistochemical adrenomedullin expression in normal normotensive pregnancies is different from that in pregnancies with pre-eclampsia. Placental tissues were obtained from seven normal normotensive pregnancies and 12 pregnancies with pre-eclampsia. The intensity of adrenomedullin staining in syncytiotrophoblasts was evaluated by means of immunohistochemistry and the ratio of the number of intact tertiary villi to that of total tertiary villi (intact/total villi ratio) was determined. The intensity of adrenomedullin expression in the placenta obtained from pregnancies with pre-eclampsia was significantly decreased compared with expression in placentas from uncomplicated normotensive pregnancies (P < 0.005). The intact/total villi ratio in placentas obtained from pregnancies with pre-eclampsia was significantly lower than that in placentas from normal normotensive pregnancies (P < 0.0001). In the amnion and extravillous trophoblast cells in both groups, no difference for the intensity of adrenomedullin expression was noted. These results suggest that adrenomedullin synthesis in the villous syncytiotrophoblasts is decreased in pregnancies with pre-eclampsia.