Placental Function in Preeclampsia

描述（由申请方提供）：炎症反应加剧和氧化应激增加是先兆子痫（PE）女性内皮功能改变的两种主要病理生理学表现，先兆子痫是人类妊娠特有的多系统疾病。虽然PE的病因尚不清楚，但据信来自胎盘滋养层（TC）的“毒性”因子进入母体循环并诱导EC功能障碍。在该项目的初始资助期间，我们确定了来源于胎盘TC的糜蛋白酶样蛋白酶（糜酶）负责EC P-选择素和E-选择素表达的上调，这是PE中EC炎症反应增加的关键分子。在这个更新补助金申请，我们将进一步研究胎盘糜酶的作用和潜在的细胞和分子机制，这种蛋白酶在调节EC功能，占EC功能改变与PE。我们的中心假设是，由胎盘TC产生的糜酶不仅在EC中启动炎症表型，而且还诱导EC氧化应激和血管激活剂释放，其特征在于PE中的EC功能障碍。这一假设将胎盘滋养层细胞畸变和EC功能障碍直接联系起来，将通过4个具体目标下概述的实验进行检验：1）表征糜酶（糜蛋白酶样蛋白酶），2）确定胎盘糜蛋白酶是否负责激活EC并诱导PE中的炎性表型，3）解决胎盘来源的糜酶是否诱导PE中的内皮氧化应激，和4）探索糜酶是否诱导刺激PE中的血管收缩的EC血管收缩剂释放。拟议的工作将采用子宫肌血管EC（UtMVEC），脐带EC，TC和VSMC。用TC或TC条件培养基培养的EC将用于研究PE期间胎盘食糜酶诱导的EC功能改变的机制。将采用EC与VSMC的共培养来研究EC和循环因子对VSMC收缩性的影响，其模拟与PE相关的血管反应。本文还将对凝乳酶诱导EC功能障碍的蛋白质、基因和信号转导途径进行研究。从拟议的工作中获得的结果应该提高我们的知识，并建立一个直接的联系，胎盘TC活动和EC功能障碍的PE。

DESCRIPTION (provided by applicant): Exaggerated inflammatory response and increased oxidative stress are the two major pathophysiological manifestations of the altered endothelial function in women with preeclampsia (PE), a multi-system disorder unique to human pregnancy. Although the etiology of PE is not known, it is believed that 'toxic' factors derived from placental trophoblasts (TCs) enter the maternal circulation and induce EC dysfunction. In the initial funding period of this project, we identified that chymotrypsin-like protease (chymase) derived from placental TCs is responsible for upregulation of EC P-selectin and E-selectin expression, key molecules of increased EC inflammatory response in PE. In this renewal grant application, we will further investigate the role of placental chymase and the potential cellular and molecular mechanisms of this protease in regulation of EC function that account for the altered EC function associated with PE. Our central hypothesis is that chymase produced by placental TCs not only initiates an inflammatory phenotype in ECs but also induces EC oxidative stress and vasoactivator release that characterize the EC dysfunction in PE. This hypothesis directly links placental trophoblast aberration and EC dysfunction, which will be tested by experiments outlined under 4 specific aims: 1) to characterize the chymase (chymotrypsin-like protease) in placentas from normal and PE pregnancies, 2) to determine if placental chymase is responsible for activating ECs and inducing an inflammatory phenotype in PE, 3) to address whether placenta-derived chymase induces endothelial oxidative stress in PE, and 4) to explore if chymase-induced EC vasoconstrictor release that stimulates vasoconstriction in PE. The proposed work will employ uterine myometric vascular ECs (UtMVECs), umbilical cord ECs, TCs, and VSMCs. ECs cultured with TC or TC conditioned medium will be used to study mechanisms underlying the placental chymase-induced alterations in EC function during PE. Co-culture of ECs with VSMCs will be employed to study effects of ECs and circulating factors on VSMC contractility that mimic a vascular response of relevance to PE. The proteins, genes and signal transduction pathways involved in chymase-induced EC dysfunction will also be studied. Results obtained from the proposed work should enhance our knowledge and establish a direct link of placental TC activity and EC dysfunction in PE.