ENDOTHELIAL ACTIVATION BY CYTOKINES AND FATTY ACID

先兆子痫是一种浆液性产科综合征，其特征是 高血压、蛋白尿和全身性水肿，影响大约 7%的孕妇。尽管它自那以来得到了承认 古代，目前对这种疾病的治疗并不是基于 对其病理生理学的理解，但仍停留在经验性的角度： 胎儿和胎盘的分娩。母婴比例偏高 与子痫前期相关的发病率和死亡率由以下原因引起 受影响妊娠的医源性早产。这个 这种综合症背后的细胞生物学仍然是一个谜。 申请的项目0002中提出的研究扩展了我们的 母体血管内皮细胞功能障碍的持续研究 先兆子痫，但有具体的重点了解 参与异常脂肪酸和前列腺素的生化途径 新陈代谢。15年多来，人们已经认识到 前列腺素在母体和胎儿组织中的异常产生 患有先兆子痫。拟议的研究将涉及 该综合征中内皮细胞激活的可能机制。二 细胞因子，即肿瘤坏死因子-α和内皮素-1，已经被博士们发现。 Conrad(Project 0005)和Taylor在子痫前期妇女血浆中的表达 由Fisher博士和McLaughlin博士(项目0003和0004)使用 胎盘缺氧的体外模型。在具体目标1中，我们将 研究这些细胞因子激活Key的能力 参与调节酶(磷脂酶A2和环合酶-2) 在内皮细胞前列腺素的产生中。归纳法， 这些酶的修饰(即磷酸化)和活性 将在人脐和蜕膜内皮细胞中进行评估。 脂肪酸结合和输送到内皮细胞的调节 白蛋白异构体的毒性是在特定目标2中解决的。 预防活性(TxPA)，一种血浆白蛋白的PI-5.6亚型，将 对其进行纯化，研究其脂结合和调节能力 细胞内脂肪酸前体的可得性 前列腺素生产。在具体目标#3中，激活 过氧化物酶体增殖物激活受体(PPAR) 将讨论细胞内脂肪酸代谢物。内皮细胞 将鉴定细胞PPAR并将其导入重组过氧化酶体 增殖反应元件(PPRE)报告构建将是 用于筛选这些核受体的前列腺素激活剂。 拟议的研究将描述通过哪些途径 细胞因子和脂肪酸激活血管内皮细胞 先兆子痫。这些分子应该会为 治疗性拮抗剂的未来发展，最终可能 导致合理的治疗和可能的预防 先兆子痫。

Preeclampsia is a serous obstetrical syndrome characterized by hypertension, proteinuria and generalized edema, that affects about 7 percent of pregnant women. Despite its recognition since antiquity, the current treatment of this disorder is not based on an understanding of its pathophysiology but remains empirical: delivery of the fetus and placenta. The high maternal and neonatal morbidity and mortality associated with preeclampsia results from the iatrogenic preterm interruption of affected pregnancies. The cellular biology that underlies this syndrome remains a mystery. The studies proposed in Project 0002 of the application extend our ongoing investigation of maternal vascular endothelial dysfunction in preeclampsia, but with the specific focus of understanding the biochemical pathways involved in abnormal fatty acid and prostanoid metabolism. For more than 15 years it has been recognized that prostaglandin production is abnormal in maternal and fetal tissues affected by preeclampsia. The proposed studies will address possible mechanisms of endothelial activation in this syndrome. Two cytokines, TNF-alpha and endothelin-1, have been identified by Drs. Conrad (Project 0005) and Taylor in the plasma of preeclamptic women and by Drs. Fisher and McLaughlin (Projects 0003 and 0004) using in vitro models of placental hypoxia. In Specific Aim #1, we will investigate the abilities of these cytokines to activate key regulatory enzymes (phospholipases A2 and cycloozygenase-2) involved in endothelial cell prostaglandin production. The induction, modification (i.e., phosphorylation) and activity of these enzymes will be assessed in human umbilical and decidual endothelial cells. Regulation of fatty acid binding and delivery into endothelial cells by albumin isoforms is addressed in specific Aim #2. Toxicity preventing activity (TxPA), a pI-5.6 isoform of plasma albumin, will be purified to study its lipid binding and its ability to modulate the availability of intracellular fatty acid precursors of prostanoid production. In Specific Aim #3 the activation of peroxisome proliferator activated receptors (PPARs) by extra- or intracellular fatty acid metabolites will be addressed. Endothelial cell PPARs will be identified and transfected recombinant peroxisome proliferator response element (PPRE) reporter constructs will be used to screen for prostanoid activators of these nuclear receptors. The proposed studies will characterize the pathways by which cytokines and fatty acids activate endothelial cells in preeclampsia. These molecules should provide ideal targets for the future development of therapeutic antagonists which ultimately could lead to the rational treatment and possible prophylaxis of preeclampsia.