Endothelial Barrier Function in Preeclampsia

增加的血管通透性是内皮功能障碍的重要组成部分，并且是一个重要的病理改变。 先兆子痫（PE）的病理生理事件。在最初的申请研究提案中， “先兆子痫中的内皮屏障功能”，一项来自正常人的内皮细胞（EC）的研究 从孕妇和PE妇女，我们发现，紊乱的EC连接蛋白血管 内皮（VE）-钙粘蛋白和紧密连接蛋白occludin是内皮细胞增殖的细胞基础。 PE中的渗透性。我们进一步证明胎盘释放的因子有能力 破坏EC连接接触并增加内皮渗透性。为了确定候选人 我们发现，在PE过程中，胎盘释放的分子诱导EC的炎症表型， 胎盘来源的糜蛋白酶样蛋白酶（CLP/糜蛋白酶）对血管生成有深远的影响， 内皮细胞在我们的初步研究中，我们观察到CLP可以破坏内皮连接蛋白， 分布并影响胎盘可溶性VEGF受体-1（sFlt-1）的产生。在这场竞争性的续约补助中 应用，我们将进一步探索CLP调节的潜在细胞和分子机制， PE中EC屏障功能。我们的中心假设是胎盘来源的CLP介导了增加的 通过改变内皮连接组装和增加sFlt-1释放， 胎盘我们将通过3个具体目标下概述的实验来验证这一假设：1）确定 胎盘来源的CLP在调节内皮屏障功能中的作用; 2）阐明胎盘来源的CLP在多大程度上影响内皮屏障功能， 胎盘源性蛋白酶诱导的内皮细胞粘附/紧密连接的分解是由 蛋白酶激活受体（PAR）在PE;和3）探讨是否增强滋养层细胞（TC）CLP 活性有助于增加sFlt-1从PE中胎盘的释放，以及涉及的机制。 拟议的工作将利用来自正常和PE妊娠的TC和EC。PAR siRNA将被 用于检测EC，以研究胎盘来源的CLP诱导EC破坏的潜在机制 与PE相关的完整性。将研究CLP对胎盘sFlt-1的影响。获得的结果 从拟议的工作应该提高目前的知识的作用，胎盘和EC功能障碍， 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.