Endothelial Barrier Function in Preeclampsia

血管通透性增加是内皮功能障碍的一个重要组成部分，也是一个重要的 子痫前期(PE)的病理生理事件。在最初应用的研究建议中 《子痫前期患者内皮屏障功能》--一项对正常来源内皮细胞的研究 从孕妇和PE妇女中，我们发现了无序的EC连接蛋白血管 内皮细胞(VE)-钙粘附素和紧密连接蛋白阻断素是内皮细胞增加的细胞基础 PE中的渗透性。我们进一步证明，从胎盘释放的因子有能力 破坏EC连接接触，增加内皮通透性。为了努力确定候选人 我们发现，在PE过程中，胎盘释放的分子可以诱导内皮细胞的炎症表型 胎盘来源的类糜蛋白酶(CLP/Chymase)对血管产生深远的影响 内皮细胞。在我们的初步研究中，我们观察到CLP可以破坏内皮细胞连接蛋白 胎盘组织中可溶性血管内皮生长因子受体-1(sFlt-1)的分布和影响。在这项相互竞争的续期拨款中 应用中，我们将进一步探索CLP调控的潜在细胞和分子机制 EC屏障在PE中的作用。我们的中心假设是胎盘来源的CLP介导了 血管内皮细胞连接组装改变和sFlt-1释放增加对PE血管通透性的影响 胎盘。我们将通过以下三个具体目标下的实验来检验这一假设：1)确定 胎盘来源的CLP在内皮屏障功能调节中的作用；2)阐明 胎盘来源的蛋白水解酶诱导内皮细胞黏附/紧密连接的解体是由 蛋白水解酶激活受体(PAR)在PE中的表达；3)探讨增强滋养层细胞(TC)CLP 活性促进胎盘sFlt-1的释放，并探讨其机制。 这项拟议的工作将利用来自正常和PE妊娠的TCS和ECs。PAR siRNA将是 用于转染内皮细胞以研究胎盘来源的CLP诱导内皮细胞破坏的机制 与体育相关的正直。我们将研究CLP对胎盘sFlt-1的影响。取得的成果 从拟议的工作中应加强目前对胎盘和EC功能障碍在 PE的发病机制。

Increased vascular permeability is an important component of endothelial dysfunction and a significant pathophysiological event in preeclampsia (PE). In the original application of the research proposal "Endothelial Barrier Function in Preeclampsia", a study of endothelial cells (ECs) derived from normal pregnant women and from women with PE, we have found that disorganized EC junction protein vascular endothelial (VE)-cadherin and tight junction protein occludin are the cellular basis of increased endothelial permeability in PE. We further demonstrated that factors released from the placenta have the ability to disrupt EC junction contacts and increase endothelial permeability. In an effort to identify candidate molecules released from the placenta that induce an inflammatory phenotype in ECs during PE, we found that placenta-derived chymotrypsin-like protease (CLP/chymase) exerts profound effects on vascular endothelium. In our preliminary studies, we observed that CLP could disorganize endothelial junction protein distribution and affect placenta soluble VEGF receptor-1 (sFlt-1) production. In this competing renewal grant application, we will further explore the potential cellular and molecular mechanisms by which CLP regulates EC barrier function in PE. Our central hypothesis is that placenta-derived CLPs mediate the increased vascular permeability in PE by altering endothelial junction assembly and by increasing sFlt-1 release from the placenta. We will test this hypothesis by experiments outlined under 3 specific aims: 1) to determine the role of placental derived CLP in regulation of endothelial barrier function; 2) to elucidate to what extent the placenta-derived protease-induced disassembly of endothelial adhesion/tight junctions is mediated by proteinase-activated receptor (PAR) in PE; and 3) to explore whether enhanced trophoblast (TC) CLP activity contributes to increased sFlt-1 release from the placenta in PE and what mechanisms are involved. The proposed work will utilize TCs and ECs derived from normal and PE pregnancies. PAR siRNA will be used to transfect ECs to study mechanisms underlying the placenta-derived CLP-induced disruption of EC integrity that are relevant to PE. The influence of CLP on placental sFlt-1 will be studied. Results obtained from the proposed work should enhance current knowledge of the role of the placenta and EC dysfunction in the pathogenesis of PE.