外泌体源性miRNA在子痫前期发病机制中的研究进展

子痫前期作为一种妊娠期特有的高血压综合征,严重威胁母婴安全,但目前尚无准确的生物标志物作为诊断依据且其病因和发病机制尚未完全阐明。外泌体是广泛存在于体液中的细胞外囊泡,携带多种蛋白质、脂质和核酸等生物活性分子,具有多种生物学功能,其所携带的miRNA具有特异性、多元性、抗降解、能被稳定检出的特点。研究发现子痫前期患者胎盘组织分泌的外泌体中有多种差异表达的miRNA,表明其可能参与了子痫前期的发生发展,可能作为子痫前期预测的潜在靶点。本文对外泌体源性miRNA的结构、功能及其与子痫前期发病机制的研究进展进行综述,旨在为子痫前期早期预测及诊治提供新思路。     

重度子痫前期患者血管内皮生长因子及其受体-1水平变化及意义

目的检测早发型和晚发型重度子痫前期患者血清和胎盘中血管内皮生长因子（vascular endothelial growth factor,VEGF）及血管内皮生长因子受体-1（soluble vascular endothelial growth factor receptors-1,sFlt-1）水平,探讨其在重度子痫前期发病中的作用机制。方法 20例早发型重度子痫前期患者为早发重度组,20例晚发型重度子痫前期患者为晚发重度组,20例正常晚期妊娠者为正常妊娠组,采用ELISA法检测3组血清VEGF及sFlt-1水平,同时应用免疫组织化学SP法检测胎盘组织中VEGF及sFlt-1的阳性表达率。结果血清VEGF水平及胎盘VEGF阳性表达率早发重度组（（27.5±3.6）ng/L和15%）、晚发重度组（（36.2±3.2）ng/L和30%）均低于正常妊娠组（（45.5±3.7）ng/L和45%）（P〈0.05）,早发重度组低于晚发重度组（P〈0.05）;血清sFlt-1水平及胎盘sFlt-1阳性表达率早发重度组（（42.2±3.2）ng/L和60%）、晚发重度组（（30.2±2.5）ng/L和45%）均高于正常妊娠组（（14.7±2.7）ng/L和10%）（P〈0.05）,早发重度组高于晚发重度组（P〈0.05）。结论 VEGF水平低表达及sFlt-1水平高表达可能与重度子痫前期的严重程度相关,在重度子痫前期发生、发展中起重要作用。     

sFlt-1、PLGF及PAPP-A在子痫前期临床诊断及预后判断中的应用

目的 研究可溶性血管内皮生长因子受体-1(sFlt-1)、胎盘生长因子（PLGF）及妊娠相关血浆蛋白-A(PAPP-A)在子痫前期临床诊断及预后判断中的应用效果。方法 选取2021年6月至2023年1月北京市大兴区人民医院收治的子痫前期孕妇151例为观察组,另选取同期进行孕检且结果正常的149名孕妇为对照组。比较两组孕妇sFlt-1、PLGF、PAPP-A水平;根据子痫前期严重程度不同将孕妇分为子痫前期轻度组和子痫前期重度组,比较子痫前期不同严重程度孕妇的sFlt-1、PLGF、PAPP-A水平;对比观察组患者不同妊娠结局一般资料、血清sFlt-1、PLGF、PAPP-A变化;进一步纳入多因素二元回归Logistic回归方程,分析影响子痫前期预后的危险因素。结果 观察组血清PLGF与PAPP-A水平均低于对照组,sFlt-1水平高于对照组,差异均有统计学意义（P<0.05）;子痫前期轻度组95例、子痫前期重度组56例,子痫前期重度组PLGF与PAPP-A水平低于子痫前期轻度组,sFlt-1高于子痫前期轻度组,差异均有统计学意义（P<0.05）;正常妊娠组117例,不良妊娠组...     

HtrA4和VE-cadherin在子痫前期中的表达及意义

目的 探讨HtrA丝氨酸肽酶4(HtrA4)和血管内皮钙黏素(VE-cadherin)在子痫前期(PE)中的水平变化情况.方法 分析该院收治的PE孕妇(根据孕周分为PE早发组和PE晚发组)及健康妊娠晚期孕妇的临床资料,收集母血和胎盘组织,采用酶联免疫吸附测定(ELISA)检测母血和胎盘组织中HtrA4和VE-cad-h...     

血清可溶性血管内皮生长因子受体1/胎盘生长因子比值作为预测子痫前期患病风险的价值

目的检测正常妊娠和子痫前期孕妇在中期妊娠时血清可溶性血管内皮生长因子受体1（sFlt-1）、胎盘生长因子（PLGF）水平,探讨母血sFlt-1/PLGF比值是否可作为预测孕妇发生子痫前期的标志物。方法用酶联免疫吸附试验（ELISA）检测200例正常妊娠妇女、57例子痫前期孕妇妊娠中期血清PLGF和sFlt-1水平,计算sFlt-1/PLGF比值。结果子痫前期组血清sFlt-1浓度[（1 385.0±178.8）ng/L]明显高于正常妊娠组[（1 087.3±122.7）ng/L]（P〈0.01）,血清PLGF浓度[（501.9±86.3）ng/L]明显低于正常妊娠组[（712.7±103.6）ng/L],（P〈0.01）;子痫前期组sFlt-1和PLGF水平呈负相关（r=-0.629,P〈0.01）,正常妊娠组sFlt-1和PLGF水平呈正相关（r=0.682,P〈0.01）。在妊娠中期,sFlt-1/PLGF比值〉1.9时,预测子痫前期的敏感性为89.5%,特异性为97.5%。结论妊娠中期血清sFlt-1水平增高伴有PLGF水平降低与子痫前期的发病密切相关,sFlt-1/PLGF比值可以作为预测子痫前期的标志物。     

滋养细胞碎片在子痫前期发病机制中的作用

子痫前期为妊娠特有疾病。目前其发病机制尚不明确,但胎盘因素参与子痫前期发病机制学说得到认可。脱落滋养细胞碎片是导致子痫前期发病的胎盘因素之一。滋养细胞碎片在1893年由Schmorl发现,随后的研究表明子痫前期患者的滋养细胞碎片的数量显著增高、性质发生改变,形成坏死滋养脱落细胞(NTD)。内皮细胞吞噬NTD后导致内皮细胞激活导致内皮功能障碍。抗磷脂体及一些免疫炎症因子IL-6、TGF-β1等在脱落滋养细胞碎片的发生发展过程中起到重要的作用。本文就近年来在子痫前期发病机制中对胎盘滋养脱落细胞的研究进展予以综述。     

血清TGF-β1、HIF-1α、sFlt-1联合检测在子痫前期患者中的临床意义

目的探讨血清血清转化生长因子β1(TGF-β1)、低氧诱导因子1α(HIF-1α)、可溶性血管内皮生长因子受体1(sFlt-1)联合检测在子痫前期患者中的临床意义。方法选择2014年3月至2015年3月该院接诊的重度子痫前期患者50例、轻度子痫前期患者50例及正常妊娠孕妇50例,比较3组孕妇血清TGF-β1、HIF-1α、sFlt-1水平及胎盘组织HIF-1α相对吸光度比值、sFlt-1mRNA和蛋白水平,同时比较轻度、重度子痫前期患者不良妊娠结局发生情况。结果重度子痫前期患者血清TGF-β1、HIF-1α、sFlt-1水平高于轻度子痫前期患者和正常妊娠孕妇(P0.05)。重度子痫前期患者HIF-1α相对吸光度比值、sFlt-1mRNA和sFlt-1蛋白水平高于轻度子痫前期患者和正常妊娠孕妇(P0.05);重度子痫前期患者不良妊娠结局总发生率高于轻度子痫前期患者(P0.05)。结论 TGF-β1、HIF-1α、sFlt-1参与了子痫前期的发生、发展,3种血清标志物联合检测可有效反映患者病情进展程度,可为判断患者预后提供可靠的依据。     

子痫前期孕妇外周血、胎盘组织中信号肽-CUB-表皮生长因子样结构域蛋白2的表达及意义

目的 探讨信号肽-CUB-表皮生长因子样结构域蛋白2(Signal peptide-CUB-EGF-like domaincontaining protein,SCUBE2)在子痫前期孕妇外周血及胎盘组织中的表达及临床意义。方法 选取2021年6月至2022年6月住院分娩的子痫前期患者58例,其中早发型子痫前期30例（发病妊娠周数<34周）,晚发型子痫前期组28例（发病妊娠周数≥34周）;另选取30例足月正常分娩孕妇作为对照组。收集入院治疗前孕妇的血清样本以及分娩时新鲜胎盘组织标本。ELISA检测三组孕妇血清SCUBE2、VEGFR2表达水平,免疫组化检测三组胎盘组织中SCUBE2的定位及表达,Western blot实验及实时荧光定量PCR(RT-qPCR)实验检测三组胎盘组织中SCUBE2、VEGFR2的表达水平。结果 免疫组化法检测显示,SCUBE2主要表达于胎盘合体滋养细胞及胎盘血管内皮细胞中,子痫前期组表达水平低于对照组,早发型子痫前期组表达低于晚发型子痫前期组;Western blot及RT-qPCR实验检测结果显示,三组孕妇胎盘中SCUBE2和VEGFR2蛋白及mR...     

子痫前期发病机制的研究进展

子痫前期(peeclampsia,PE)是妊娠期特有的一种疾病,是导致孕产妇和围产儿发病和死亡的主要原因之一。PE受多种因素影响,涉及多个发病环节,引起多个器官、系统受累,但其发病机制尚不清楚。研究表明,胎盘在PE的发病中占重要地位,胎盘化不足、释放大量胎盘因子是导致母体和胎儿症状的主要原因。该文主要从胎盘界面免疫失衡、低氧、血栓炎症、氧化应激、抗血管紧张素Ⅱ1型受体抗体、血管生成失衡、过度的全身性炎症反应和内皮细胞功能紊乱在PE中的发病机制进行综述。     

子痫前期患者胎盘组织及外周血中胎盘生长因子及可溶性血管内皮生长因子受体1的表达及意义

目的 研究子痫前期患者胎盘生长因子(placental growth factor, PlGF)、可溶性血管内皮生长因子受体1(soluble vascular endothelial growth factor receptor 1, sFlt-1)的mRNA及蛋白质在胎盘组织中的表达及外周血中的浓度水平, 探讨其与子痫前期的关系。 方法 选取57例子痫前期患者（轻度子痫前期31例、重度子痫前期26例）和60例健康孕妇,用半定量RT-PCR及western blot分别检测胎盘组织中sFlt-1、PlGF的mRNA及蛋白质的表达水平, ELISA法检测血清中PlGF和sFlt-1水平。 结果 轻、重度子痫前期患者组胎盘组织PlGF mRNA、蛋白质表达水平以及血清PlGF浓度均显著低于健康孕妇组（t分别为14.22、21.80、12.10、15.17、7.14、10.18,P均<0.01）;而胎盘组织sFlt-1 mRNA、蛋白质表达水平以及血清sFlt-1浓度显著高于健康孕妇组（t分别为12.43、17.06、13.70、18.84、13.55、15.19,P均<0.01）;轻、重度子痫前期组外周血PlGF和sFlt-1之间呈显著负相关（r=-0.49, r=-0.53,P<0.05）。 结论 子痫前期胎盘组织中PlGF水平降低伴sFlt1水平升高与子痫前期的发病密切相关。     

Changes in mRNA and protein levels of human HtrA1, HtrA2 and HtrA3 in ovarian cancer

ObjectivesExpression of human HtrA1, HtrA2, HtrA3 and TGF-β1 genes was examined in ovarian tissue specimens including 19 normal ovaries, 20 benign tumors, 7 borderline tumors, 44 cancers and 8 Krukenberg tumors.Design and methodsmRNA and protein levels were evaluated by semi-quantitative RT-PCR and Western-blotting methods, respectively.ResultsA statistically significant decrease of HtrA1 and HtrA3 expression in ovarian tumors comparing to normal tissues was observed. A dramatic decrease of HtrA3 mRNA and protein levels in all tumor tissue groups, and a loss of HtrA3 protein in 30% malignant tumors were found. A significant decrease of HtrA1 mRNA, and of HtrA3 mRNA and protein in malignant tumors compared to benign tumors was revealed. HtrA2 expression in tumor tissues was slightly decreased. Expression of TGF-β1 in tumor tissues was not significantly different compared to control tissues.ConclusionsOur results show downregulation of HtrA1 and HtrA3 genes' expression in different types of ovarian tumors and give additional evidence that these genes may function as tumor suppressors.     

急性髓系白血病中HtrA2和WT1基因的表达

本研究通过检测HtrA2和WT1基因在急性髓系白血病(AML)患者骨髓单个核细胞及细胞系中的表达,探讨其表达水平与临床变量的关系及二者之间的相关性。应用RQ-PCR方法检测104例初诊AML患者的HtrA2和WT1基因表达水平;比较其表达量与正常对照的差别,并分析与年龄、性别、白细胞计数、诊断分型、预后分型及治疗疗效的关系;比较治疗前后表达量的变化,分析HtrA2与WT1二者相关性。结果表明:AML患者骨髓细胞中HtrA2表达量显著低于正常对照组(P<0.01),而WT1则显著高于正常对照组(P<0.01);二者表达量与患者年龄、性别、白细胞计数无关;HtrA2在不同的NCCN预后分组中表达量无显著差异,而WT1在预后良好组表达量明显低于预后中等组(P=0.003);无论完全缓解(CR)组还是非完全缓解(non-CR)组HtrA2表达量在治疗后均显著高于治疗前(P<0.05),而WT1表达量只有在CR组治疗后才低于治疗前(P<0.01),non-CR组治疗后虽然也低于治疗前,但差异无统计学意义;HtrA2与WT1的表达水平呈弱负相关(r=-0.249,P=0.011)。结论:HtrA2和WT1表达与白血病的发生、发展密切相关,二者表达水平呈负相关,上调HtrA2基因的表达和干扰WT1基因的表达有望成为白血病治疗的靶点。     

初治淋巴瘤患者血浆中VEGF-C、VEGF-D、VEGFR-2及VEGFR-3的表达水平及临床意义

本研究检测初治淋巴瘤患者血浆中血管生成相关因子VEGF-C、VEGF-D、VEGFR-2、VEGFR-3的表达水平,探讨其与淋巴瘤临床特点及预后的相关性,寻找指导淋巴瘤抗血管靶向治疗及判断预后的有价值的指标和因素。应用酶联免疫吸附测定的方法检测86例初治淋巴瘤患者血浆中VEGF-C、VEGF-D、VEGFR-2、VEGFR-3的表达水平。结果表明,多因素分析结果显示,VEGF-C在非霍奇金淋巴瘤患者中呈低表达水平,而在霍奇金淋巴瘤患者中表达水平较高;VEGFR-2在年龄>60岁淋巴瘤患者中表达较高;VEGF-D在IPI>2分的患者组中表达较低。单因素分析显示,VEGF-D在IPI>2分患者组中呈低水平表达;VEGF-D、VEGF-C在无B症状组高水平表达。因子间的关联性分析表明,VEGF-D与VEGFR-2、VEGFR-3的表达水平呈正相关。结论:VEGF-C、VEGF-D、VEGFR-2、VEGFR-3在淋巴瘤的发病中具有重要作用,可作为细化指导淋巴瘤抗血管靶向治疗、了解病情、判断预后的指标。     

Assessing the activity of cediranib, a VEGFR-2/3 tyrosine kinase inhibitor, against VEGFR-1 and members of the structurally related PDGFR family

Cediranib is a potent inhibitor of the VEGF receptor (VEGFR)-2 and VEGFR-3 tyrosine kinases. This study assessed the activity of cediranib against the VEGFR-1 tyrosine kinase and the platelet-derived growth factor receptor (PDGFR)-associated kinases c-Kit, PDGFR-α, and PDGFR-β. Cediranib inhibited VEGF-A-stimulated VEGFR-1 activation in AG1-G1-Flt1 cells (IC(50) = 1.2 nmol/L). VEGF-A induced greatest phosphorylation of VEGFR-1 at tyrosine residues Y1048 and Y1053; this was reversed by cediranib. Potency against VEGFR-1 was comparable with that previously observed versus VEGFR-2 and VEGFR-3. Cediranib also showed significant activity against wild-type c-Kit in cellular phosphorylation assays (IC(50) = 1-3 nmol/L) and in a stem cell factor-induced proliferation assay (IC(50) = 13 nmol/L). Furthermore, phosphorylation of wild-type c-Kit in NCI-H526 tumor xenografts was reduced markedly following oral administration of cediranib (≥1.5 mg/kg/d) to tumor-bearing nude mice. The activity of cediranib against PDGFR-β and PDGFR-α was studied in tumor cell lines, vascular smooth muscle cells (VSMC), and a fibroblast line using PDGF-AA and PDGF-BB ligands. Both receptor phosphorylation (IC(50) = 12-32 nmol/L) and PDGF-BB-stimulated cellular proliferation (IC(50) = 32 nmol/L in human VSMCs; 64 nmol/L in osteosarcoma cells) were inhibited. In vivo, ligand-induced PDGFR-β phosphorylation in murine lung tissue was inhibited by 55% following treatment with cediranib at 6 mg/kg but not at 3 mg/kg or less. In contrast, in C6 rat glial tumor xenografts in mice, ligand-induced phosphorylation of both PDGFR-α and PDGFR-β was reduced by 46% to 61% with 0.75 mg/kg cediranib. Additional selectivity was showed versus Flt-3, CSF-1R, EGFR, FGFR1, and FGFR4. Collectively, these data indicate that cediranib is a potent pan-VEGFR kinase inhibitor with similar activity against c-Kit but is significantly less potent than PDGFR-α and PDGFR-β.     

An antibody targeted to VEGFR-2 Ig domains 4-7 inhibits VEGFR-2 activation and VEGFR-2-dependent angiogenesis without affecting ligand binding

Inhibition of VEGFR-2 signaling reduces angiogenesis and retards tumor growth. Current biotherapeutics that inhibit VEGFR-2 signaling by either sequestering VEGF ligand or inhibiting VEGF binding to VEGFR-2 may be compromised by high VEGF concentrations. Here we describe a biotherapeutic that targets VEGFR-2 signaling by binding to Ig domains 4-7 of VEGFR-2 and therefore has the potential to work independently of ligand concentration. 33C3, a fully human VEGFR-2 antibody, was generated using XenoMouse technology. To elucidate the mechanism of action of 33C3, we have used a number of competition and binding assays. We show that 33C3 binds VEGFR-2 Ig domains 4-7, has no impact on VEGF-A binding to VEGFR-2, and does not compete with an antibody that interacts at the ligand binding site. 33C3 has a high affinity for VEGFR-2 (K(D) < 1 nmol/L) and inhibits VEGF-A induced phosphorylation of VEGFR-2 with an IC(50) of 99 ± 3 ng/mL. In vitro, in a 2D angiogenesis assay, 33C3 potently inhibits both tube length and number of branch points, and endothelial tubule formation in a 3D assay. In vivo, 33C3 is a very effective inhibitor of angiogenesis in both a human endothelial angiogenesis assay and in a human skin chimera model. These data show targeting VEGFR-2 outside of the ligand binding domain results in potent inhibition of VEGFR-2 signaling and inhibition of angiogenesis in vitro and in vivo.     

Effects of magnesium sulphate on placental expression of endothelin 1 and its receptors in preeclampsia

OBJECTIVES: To investigate the effects of magnesium sulphate (MgSO(4)) on placental expression of endothelin 1 (ET-1) and its receptors in preeclampsia (PE). DESIGN AND METHODS: Placentas were obtained from 10 normotensive (NT group) and 18 moderate preeclamptic (PE group) women. Among the PE group, 10 patients were treated with 0.9% NaCl solution (PES) and 8 women received MgSO(4) (PEMgSO(4)). Placental mRNAs of ET-1, ET-1(A) receptor (ET-1(A)R) and ET-1(B) receptor (ET-1(B)R) were evaluated by Northern blot and quantified using densitometry. RESULTS: Placental ET-1(B)R expression was lower (P<0.05) in the PES group without significant changes in the mRNAs of ET-1 and ET-1(A)R when compared with the NT group. MgSO(4) treatment was associated with decreased ET-1 and increased ET-1(B)R (P<0.05) expression, without significant changes in ET-1(A)R. CONCLUSIONS: The results of the present study showed that moderate PE is associated with low placental expression of ET-1(B)R, and MgSO(4) treatment resulted in placental expression changes of the ET-1/receptors system.     

缺氧、VEGF及其受体表达在高血压肾损伤中的作用

目的:探讨缺氧、VEGF及其受体表达在高血压肾损伤中的作用。方法:将大鼠分为3组:①假手术组;②两肾一夹高血压未治疗组;③两肾一夹高血压降压治疗组。每组8只,观察10周后处死,对未钳夹的右肾进行常规病理、RT—PCR及免疫组化检测。用缺氧探针显示肾组织缺氧情况。结果:高血压组血清肌酐与假手术组比差异无统计意义(P＞0．05),但血压、24h尿蛋白定量、肾小球损伤指数、微动脉壁/腔比均较假手术组有上升(P＜0．01)。假手术组大鼠未钳夹肾髓质有轻度缺氧,但皮质无明显缺氧表现。高血压组髓质和皮质均有明显缺氧(P＜0．01),主要阳性部位在肾小管和病变较严重的肾小球。皮质VEGF、VEGFR-2表达均较假手术组明显升高(P＜0．01),阳性部位与缺氧探针阳性部位分布一致。与未治疗组比较,降压治疗组的血压、24h尿蛋白定量、肾小球损伤指数、肾皮质缺氧状况均有所改善,VEGF、VEGFR-2表达显著下调(P＜0．05)。结论:两肾一夹高血压大鼠模型可作为慢性肾脏缺氧的动物模型。高血压大鼠的肾皮质VEGF及其受体表达上调,并与缺氧部位分布一致,这可能是对组织缺氧的代偿反应,并参与了慢性高血压肾损伤的发生。     

Ta1722, an anti-angiogenesis inhibitor targeted on VEGFR-2 against human hepatoma

In order to investigate the anti-angiogenesis potential and related mechanisms of Ta1722 (a novel taspine derivative compound), a series of experiments in vivo and in vitro were carried out. The proliferation on human cell lines of SMMC-7721, A549, MCF-7, Lovo, and ECV304 was examined by MTT. Angiogenesis inhibition was examined by chick embryo chorioallantoic membrane (CAM) angiogenesis and tube formation assays. Related angiogenesis proteins and their mRNA expression were determined by western blotting and RT-PCR. In addition, the SMMC-7721 nude mouse xenotransplant model was used to evaluate the inhibition of tumor growth. The results showed that Ta1722 inhibited cell proliferation, angiogenesis of CAM and tube formation, and downregulated related positive angiogenesis proteins. The above indicated Ta1722 could serve as a promising candidate of angiogenesis inhibitors by interrupting the VEGF/VEGFR-2 pathway.