雌激素受体α在子痫前期胎盘的表达及临床意义

目的:通过比较雌激素受体α(ERα)在子痫前期胎盘和正常胎盘中组织细胞的定位,探讨ERα与子痫前期疾病的相关性及其临床意义。方法:用免疫组化SP法检测28例正常足月妊娠胎盘组织和33例子痫前期胎盘组织中ERα的表达。结果:免疫组织化学结果显示两组的ERα均呈阳性表达,子痫前期组的ERα在胎盘细胞滋养细胞的细胞质内或细胞核内的阳性表达率高于正常孕妇组(P<0.05)。结论:免疫组化检查定位于胎盘绒毛细胞滋养细胞、绒毛毛细血管管壁细胞和绒毛间质纤维组织成纤维细胞的ERα作为实现雌激素功能的介导中枢,可能通过调控雌激素的作用参与了子痫前期的发生和发展过程。     

α7 nAChR和sFlt-1mRNA在重度子痫前期患者胎盘的表达

目的:检测α7烟碱乙酰胆碱受体(alpha 7 nicotini cacetylcholine receptor,α7nAChR)和可溶性血管内皮生长因子受体-1(soluble fms-like tyrosine kinase-1,sFlt-1)在重度子痫前期患者胎盘组织中mRNA的表达水平,探讨它们在子痫前期发生发展中的作用。方法:用半定量逆转录聚合酶链反应(RT-PCR)技术检测18例重度子痫前期患者和12例正常妊娠孕妇的胎盘。结果:(1)α7nAChR和sFlt-1mRNA在正常孕妇和重度子痫前期患者胎盘组织中均有表达。(2)重度子痫前期患者α7nAChR和sFlt-1mRNA的表达水平上调,且两者上升水平呈正相关。结论:胎盘组织中α7nAChR和sFlt-1mRNA的过度表达可能与子痫前期发病有关,参与了子痫前期的病理生理过程。     

The immunohistochemical evaluation of VEGF in placenta biopsies of pregnancies complicated by preeclampsia

Objective The study was designed to determine the protein levels of vascular endothelial growth factor (VEGF) in the placenta biopsies of patients with preeclampsia and compare with normal controls. Design Prospective cohort study. Methods The placental biopsies were obtained from ten patients with preeclampsia and ten patients of control group at the time of delivery. Avidin–biotin-peroxidase immunohistochemistry was then performed to identify levels of VEGF protein within the tissue and a semi-quantitative method was devised to score the amount of staining present in the sample. Two histopathologists who were blinded to the groups were asked to score each sample for the intensity of staining and the number of cells stained in a randomly selected per high-power fields of each sample. The resulting “H-score” was computed as a product of intensity and percent of cells stained. Results The VEGF expression was significantly higher in placenta biopsies of preeclamptic patients compared to that of controls (271.2 ± 22.65 vs. 201.9 ± 12.33, P = 0.000). Conclusion Immunostaining of VEGF is significantly higher in placenta biopsies of patients with preeclampsia.     

子痫前期孕妇血清与胎盘尿紧张素Ⅱ的表达及意义

目的:检测子痫前期孕妇外周血和新生儿脐血血清中尿紧张素Ⅱ(urotensinⅡ,UⅡ)水平和UⅡmRNA在胎盘组织的表达水平,探讨其在子痫前期发生发展中的作用。方法:(1)ELISA测定30例子痫前期患者及15例正常晚期妊娠孕妇(对照组)外周血和新生儿脐血血清UⅡ水平;(2)用RT-PCR法检测各组孕妇胎盘组织中UⅡmRNA的表达水平。结果:(1)重度子痫前期组孕妇外周血血清UⅡ水平明显高于轻度子痫前期组和对照组,差异均有显著性(P<0.05)。轻度和重度子痫前期组的脐静脉血清UⅡ浓度均明显高于对照组(P<0.05,P<0.001)。(2)胎盘组织UⅡmRNA表达水平在轻度和重度子痫前期组均明显高于对照组,差异有显著性(P<0.05)。结论:UⅡ可能参与子痫前期全身小血管痉挛的机制并在胎盘组织缺血缺氧和动脉粥样硬化的发生起重要作用。     

细胞外基质金属蛋白酶诱导因子在子痫前期及子痫患者胎盘组织中表达的研究

目的探讨细胞外基质金属蛋白酶诱导因子(EMMPRIN)在子痫前期及子痫患者胎盘组织中的表达及其与子痫前期及子痫发病的关系。方法采用免疫组化链霉菌抗生物素蛋白-过氧化酶连接法和RT-PCR技术检测不同孕周的44例子痫前期(重度)孕妇(子痫前期组)、38例子痫孕妇(子痫组)和49例正常孕妇(正常妊娠组)胎盘组织中EMMPRIN蛋白及mRNA的表达。结果(1)EMMPRIN蛋白阳性表达：子痫前期组EMMPRIN蛋白表达中度阳性率为18%(8/44),强阳性率为9%(4/44);子痫组EMMPRIN蛋白中度阳性率为21%(8/38),强阳性率为13%(5/38),两组分别比较,差异均无统计学意义(P＞0．05)。正常妊娠组EMMPRIN蛋白表达中度阳性率为12%(6/49),强阳性率为82%(40/49),与子痫前期组及子痫组比较,差异均有统计学意义(P＜0．001)。(2) EMMPRIN mRNA表达：子痫前期组孕晚期(37～40周)EMMPRIN mRNA为0．342±0．002,子痫组为0．344±0．023,两组比较,差异均无统计学意义(P＞0．05)。正常妊娠组孕晚期(37～40周) EMMPRIN mRNA为0．872±0．094,分别与子痫前期组及子痫组比较,差异均有统计学意义(P＜0．001)。结论子痫前期及子痫患者胎盘组织中EMMPRIN表达水平下降是子痫前期及子痫发病的重要原因;EMMPRIN可作为子痫前期及子痫发病的一个预测指标。     

人类白细胞相关抗原-G及其各亚型mRNA在正常妊娠及重度子前期患者胎盘组织中的表达

目的研究人类白细胞相关抗原-G(HLA-G)及其各个亚型mRNA在正常妊娠及重度子前期患者胎盘组织中的表达。方法选取2005-01-2005-06第四军医大学唐都医院10例重度子前期患者(研究组)及10例正常妊娠胎盘组织(对照组),应用荧光定量RT-PCR比较两组间HLA-G及其各个亚型mRNA(HLA-G1、G2、G3、G4、G5、G6)的表达。结果与对照组相比,重度子前期患者胎盘组织中总HLA-GmRNA显著降低,以HLA-G1、HLA-G3降低为主,差异有统计学意义(P<0.05)。结论HLA-G1与HLA-G3在胎盘组织中的低表达可能与妊娠期高血压疾病的发病和病理生理过程有关。     

膜型基质金属蛋白酶表达与妊娠疾病的关系

目的 :研究葡萄胎和先兆子痫等与滋养层细胞密切相关的妊娠疾病在病理发生机制上与膜型基质金属蛋白酶表达的关系。方法 :采用逆转录多聚酶链反应 (RT PCR)检测方法 ,比较葡萄胎组织、早孕绒毛、先兆子痫足月胎盘和正常足月胎盘 (自然分娩和剖宫产 )组织中膜型基质金属蛋白酶 (MT MMP)的表达情况。结果 :5种胎盘绒毛组织均表达MT2 MMP和MT5 MMP。在相对表达量上 ,MT2 MMP及MT3 MMP在正常早孕绒毛组织中的表达强于葡萄胎组织 ,正常足月胎盘组织中MT2 MMP和MT5 MMP的表达也强于先兆子痫患者胎盘组织中 (P <0 .0 5 )。而自然分娩和剖宫产足月胎盘绒毛组织中MT2 MMP、MT3 MMP和MT5 MMP的表达差异均无显著性 (P >0 .0 5 )。结论 :MT2 MMP、MT3 MMP和MT5 MMP等膜型MMP在胎盘绒毛组织中的表达差异 ,与葡萄胎、先兆子痫等和滋养层密切相关的妊娠疾病的病理发生有关     

人类白细胞相关抗原-G及其各亚型mRNA在正常妊娠及重度子痫前期患者胎盘组织中的表达

目的 研究人类白细胞相关抗原-G（HLA-G）及其各个亚型mRNA在正常妊娠及重度子痫前期患者胎盘组织中的表达.方法 选取2005-01-2005-06第四军医大学唐都医院10例重度子痫前期患者（研究组）及10例正常妊娠胎盘组织（对照组）,应用荧光定量RT-PCR比较两组间HLA-G及其各个亚型mRNA（HLA-G1、G2、G3、G4、G5、G6）的表达.结果 与对照组相比,重度子痫前期患者胎盘组织中总HLA-G mRNA显著降低,以HLA-G1、HLA-G3降低为主,差异有统计学意义（P＜0.05）.结论 HLA-G1与HLA-G3 在胎盘组织中的低表达可能与妊娠期高血压疾病的发病和病理生理过程有关.     

Elevation of granulocyte-macrophage colony-stimulating factor in the placenta and blood in preeclampsia

OBJECTIVE: This study investigated whether granulocyte-macrophage colony-stimulating factor (GM-CSF) levels in the placenta and blood in preeclampsia differed from those in normal pregnancies. Macrophage colony-stimulating factor (M-CSF) levels in the placenta were also measured. STUDY DESIGN: The subjects were 44 pregnant women carrying single fetuses, of whom 22 were women with normal pregnancies and 22 were women with preeclampsia. Their average gestational age at entry was 37 to 38 weeks of gestation. Peripheral blood was collected before the onset of labor. Separated serum was obtained after centrifugation and stored at -20 degrees C. A tissue segment of the placenta was cut immediately after delivery. The frozen placental tissue was placed into a plastic tube containing phosphate-buffered saline solution. The tissue was fully homogenized and then centrifuged. Separated supernatant was frozen at -80 degrees C for subsequent determination. GM-CSF levels in separated serum were measured, and GM-CSF, M-CSF, and total protein (TP) levels in separated supernatant were also measured. RESULTS: Not only GM-CSF levels in blood but also GM-CSF/TP levels in the placenta were significantly higher (P<.05) in preeclampsia than in normal pregnancies. Similar results were obtained for M-CSF/TP levels in the placenta. CONCLUSIONS: We demonstrated a significant increase in placenta levels of GM-CSF/TP in preeclampsia. Elevated GM-CSF in the placenta may be related to immunologic abnormalities contributing to the etiology of preeclampsia.     

Altered mRNA expression of ecNOS and iNOS in myometrium and placenta from women with preeclampsia

Objective: Preeclampsia is a syndrome involving dysfunction of vascular endothelium and imbalance between endothelium derived constricting and relaxing factors. Recent evidence suggests that endothelium-derived nitric oxide (NO) plays a role in the regulation of vascular resistance during normal pregnancy and preeclampsia. NO is a potent vasodilator and is generated by the catalytic action of nitric oxide synthases ecNOS and iNOS in myometrium and placenta. Methods: In this study mRNA expressions of ecNOS and iNOS were compared in myometrium and placenta. Biopsies were collected from women with preeclampsia (n=8) and normal pregnancies (n=12). ecNOS and iNOS mRNA levels were determined using RT-PCR and expressed as arbitrary units after correction for control GAPDH gene mRNA levels. Results: The mRNA expression of ecNOS was significantly higher in both myometrium (p<0.05) and placenta (p<0.05) from women with preeclampsia compared to that in normal pregnancies, while the iNOS mRNA level was not altered in myometrium and lower in placenta (p<0.05) from women with preeclampsia. Conclusions: The higher ecNOS mRNA expression might be a compensatory response to an impaired vasodilatation in the uteroplacental circulation during preeclampsia. Whether the similar and reduced levels of iNOS mRNA expression in myometrium and placenta, respectively, in women with preeclampsia is of importance remains to be further evaluated. Received: 30 October 2000 / Accepted: 30 October 2000     

Decreased expression of complement 3a receptor (C3aR) in human placentas from severe preeclamptic pregnancies

Objectives

The aim of this study was to determine the expression of the anaphylatoxin receptors complement C3a receptor (C3aR) and C5a receptor (C5aR) in the placentas of pregnancies complicated by severe early onset preeclampsia.

Study design

We recruited women with pregnancies complicated by severe early-onset preeclampsia (n = 19, 11 of which were further complicated with IUGR) and women with preterm pregnancies not affected by preeclampsia (n = 8). Gene and protein expression of C3aR and C5aR was analysed by quantitative RT-PCR and Western blotting, respectively.

Results

C3aR was detected in the Hofbauer cells in the villous stroma of the placenta. C5aR staining was detected in the syncytiotrophoblast and endothelial cells. We found significantly decreased expression of C3aR mRNA and protein expression in placentas with preeclampsia compared to controls. However, C5aR expression was not significantly different between preeclamptic and control placentas at either the mRNA or protein level.

Conclusions

Decreased C3aR expression indicates a dysregulation of the complement system in the placentas of preeclamptic women. Further studies would elucidate the exact mechanisms that complement has in preeclampsia.     

长链脂肪酸氧化酶mRNA在正常妊娠及重度子痫前期胎盘中的表达

目的研究长链羟酰基辅酶A脱氢酶(long-chain3-hydroxyacyl-CoAdehydrogenase,LCHAD)在正常妊娠不同孕期绒毛或胎盘组织的表达情况以及在伴有肝脏损害和不伴有肝脏损害重度子痫前期胎盘的表达差异。方法应用原位杂交和RT-PCR方法对早孕绒毛组织(10例)、妊娠中期胎盘组织(10例)、正常妊娠晚期胎盘组织(10例)及32例重度子痫前期胎盘组织进行LCHAD基因的定位表达及半定量测定。结果原位杂交实验显示正常妊娠早、中、晚期绒毛或胎盘组织及重度子痫前期胎盘组织滋养细胞中存在LCHAD阳性表达。RT-PCR实验显示①妊娠早期绒毛LCHAD表达与妊娠中期比较,P=0.844;妊娠早期绒毛LCHAD表达高于晚期胎盘,P=0.020;妊娠中期胎盘LCHAD表达也高于晚期胎盘P=0.026;②发病孕周≤34周早发型重度子痫前期伴肝损害胎盘组织中LCHAD表达均值为(0.449±0.038),不伴肝损害LCHAD表达均值为(0.482±0.042),伴肝损害较不伴肝损害者表达有减弱,但两组比较,P=0.084;发病孕周≤34周早发型重度子痫前期伴肝损害LCHAD表达量与正常晚期比较,P=0.05,而不伴肝损害重度子痫前期LCHAD表达量与正常晚期比较,P=0.775。结论本研究显示在妊娠的早中晚期滋养细胞中均存在长链脂肪酸氧化代谢,妊娠早中期LCHAD的mRNA表达高于妊娠晚期;早发型重度子痫前期伴肝损害胎盘组织中LCHAD表达均值与不伴有肝损害者比较虽无统计学差异,但是有明显降低趋势。提示长链脂肪酸氧化代谢对子痫前期伴发肝脏损害的影响还有待酶活性和蛋白水平以及代谢调节方面的深入研究。     

Activation of the cholinergic anti-inflammatory pathway by nicotine ameliorates lipopolysaccharide-induced preeclampsia-like symptoms in pregnant rats

IntroductionPreeclampsia (PE) exerts a more intense systemic inflammatory response than normal pregnancy. Recently, the role of the cholinergic anti-inflammatory pathway (CAP) in regulating inflammation has been extensively studied. The aim of this study was to investigate the effect of nicotine, a selective cholinergic agonist, on lipopolysaccharide (LPS)-induced preeclampsia-like symptoms in pregnant rats and to determine the molecular mechanism underlying it.MethodsRats were administered LPS (1.0 μg/kg) via tail vein injection on gestational day 14 to induce preeclampsia-like symptoms. Nicotine (1.0 mg/kg/d) and α-bungarotoxin (1.0 μg/kg/d) were injected subcutaneously into the rats from gestational day 14–19. Clinical symptoms were recorded. Serum and placentas were collected to determine cytokine levels using Luminex. The mRNA and protein expression levels of α7 nicotinic acetylcholine receptor (α7nAChR) were determined using Real time-PCR and Western blot analysis. Immunohistochemistry was performed to determine the level of activation of nuclear factor-κB (NF-κB) in placentas.ResultsNicotine significantly ameliorated LPS-induced preeclampsia-like symptoms in pregnant rats (P < 0.05). Nicotine treatment decreased the levels of LPS-induced pro-inflammatory cytokines in the serum (P < 0.05) and placenta (P < 0.05). Nicotine significantly increased the expression of α7nAChR (P < 0.01) and attenuated the activation of NF-κB p65 in the placenta in LPS-induced preeclampsia (P < 0.01). Meanwhile, these protective effects of nicotine were abolished by the administration of the cholinergic antagonist α-bungarotoxin in preeclampsia rats.DiscussionOur findings suggest that the activation of α7nAChR by nicotine attenuates preeclampsia-like symptoms, and this protective effect is likely the result of the inhibition of inflammation via the NF-κB p65 pathway.     

子(癎)前期患者胎盘组织中Toll样受体2和Toll样受体4的表达及意义

目的研究子痫前期（PE）患者胎盘组织中Toll样受体2（TLR2）和Toll样受体4（TLR4）的表达及其与胎盘中肿瘤坏死因子α（TNF-α）、白介素6（IL-6）表达的关系,探讨PE胎盘中Toll样受体变化的意义。方法2005年1月至2006年12月在哈尔滨医科大学第一临床医学院采用免疫组化和RT-PCR法检测PE患者和正常晚期妊娠妇女胎盘组织中TLR2和TLR4的定位及表达,同时用脂多糖（LPS）体外刺激胎盘组织中的TLR4,检测其上清液中TNF-α、IL-6的变化。其中PE患者44例（PE组）,同期正常晚期妊娠妇女22例（对照组）。结果PE组和对照组胎盘组织中均有TLR2和TLR4的表达,其中PE组TLR4表达明显比对照组增强,组间差异有显著性意义（P〈0.05）;LPS体外刺激PE组胎盘组织,导致TNF-α、IL-6分泌量增加,刺激后2hPE组TNF-α、IL-6分泌量与对照组比较差异无显著意义（P〉0.05）,刺激后6hPE组TNF-α、IL-6的分泌量升高明显,与对照组比较,差异有显著性意义（P〈0.01）。结论PE患者胎盘中TLR4表达增加与TNF-α、IL-6改变密切相关,是造成局部细胞因子微环境异常的主要因素,其增加可能是PE发病机制中的关键环节之一。     

Placental mRNA expression of angiopoietins (Ang)-1, Ang-2 and their receptor Tie-2 is altered in pregnancies complicated by preeclampsia

Objective

To investigate the placental expression of angiopoietin (Ang)-1, Ang-2 and their receptor, Tie-2, in preeclampsia (PE) with or without intrauterine growth restriction (IUGR).

Methods

Case-control study including placentas from 28 PE pregnancies, 30 PE-IUGR pregnancies and 40 controls. The expression status of the genes was evaluated by quantitative real-time PCR.

Results

In both PE and PE-IUGR groups, compared to the control group, there was significantly higher expression of Ang-2 (p < 0.001) and Tie-2 (p = 0.008) and lower expression of Ang-1 (p = 0.001). The magnitude of the difference was similar for Ang-1 for both groups, whereas the magnitude of the differences was higher for Ang-2 and Tie-2 in PE-IUGR group compared to controls. Ang-2 and Tie-2 were correlated in both PE (r = 0.8602, p < 0.001) and PE-IUGR (r = 0.6342, p < 0.001) groups. In PE-IUGR group, Ang-1 was associated to Ang-2 (r = 0.3458, p = 0.0452) and Tie-2 (r = 0.4448, p = 0.0084). Log₁₀Ang-1 but not Ang-2 was gestational age dependent (R2 = 0.40, p < 0.001). After conversion in Multiples of the Median (MoM) log₁₀ MoM Ang-1 was reduced in the PE group (mean = −0.8181, p < 0.001) and the PE-IUGR group (mean = −1.2583, p < 0.001) compared to control group (mean = −0.0924).

Discussion

We have demonstrated increased placental expression of Ang-2 and Tie-2 along with lower expression levels of Ang-1 in pregnancies with PE and PE-IUGR.

Conclusion

The angiopoietin axis seems to be disrupted in PE pregnancies. Whether the results of this study represent the angiogenic imbalance observed in PE pregnancies or they are part of the pathophysiology of this condition has to be further investigated.     

miR-223-3p靶向NLRP3调控子痫前期胎盘中炎性反应的机制研究

目的:探讨miR-223-3 p能否通过靶向NLRP3参与调节子痫前期(PE)胎盘中的炎性反应.方法:选取2018年12月至2020年01月在江苏省苏北人民医院住院行剖宫产分娩的60例孕妇,其中PE组30例,正常对照组30例.采用实时定量聚合酶链式反应(RT-qPCR)、蛋白免疫印迹、免疫组化法对胎盘组织中NLRP3和...