Luteolin as a therapeutic for hypertension in pregnancy

项目概要/摘要 子痫前期（PE）是一种常见的、危及生命的妊娠期高血压疾病。这种疾病影响 几乎占所有怀孕的7%，是孕产妇和胎儿发病率和死亡率的主要原因。有 除了早期分娩外，目前没有治疗PE的疗法。我们实验室和其他机构的数据表明， PE患者存在血管生成失衡（过量可溶性fms样酪氨酸激酶1 [sFlt 1]），炎症证据 （过量的肿瘤坏死因子TNF-α、白细胞介素如IFN-γ诱导蛋白10-IP-10和IL-6）和 过量的活性氧（ROS）。这些可溶性因子从胎盘释放到 血流干扰促血管生成因子如胎盘生长因子（PlGF）的供应， 一氧化氮（NO）减少，内皮素-1（ET-1）增加，最终导致内皮功能障碍， 高血压PE缺乏治疗选择可能是由于其复杂的病因和药物的局限性 怀孕期间使用。为这种情况确定一种安全的治疗方法是对健康的一个主要的未满足的需求。 孕妇化学文库和高通量筛选方法的可用性使得 研究人员发现新的分子，干扰信号通路参与疾病生物学。我们 筛选了一个天然化合物的化学库，并确定了一种有效的抑制剂，即木犀草素 sFlt 1在人胎盘细胞系和胎盘外植体中的表达。此外，我们发现毛地黄黄酮抑制了 多种促炎细胞因子（如IL-1、IL-27、IL-6和IP-10），并干扰不同的细胞因子， 抑制PE发病机制中涉及的几种关键蛋白质的途径。具体来说，我们发现毛地黄黄酮是 低氧诱导因子1α（HIF-1α，sFlt 1的主要调节因子）的有效抑制剂。此外，毛地黄黄酮减少 TNF- α在体外诱导ET-1产生，降低血压，舒张子宫动脉，改善胎盘 胎盘缺血大鼠PE模型灌流。我们令人兴奋和新颖的初步数据使我们提出了 中心假设毛地黄黄酮改善子宫灌注并降低总外周阻力， 通过减少胎盘sFlt 1和HIF-1α的产生以及抑制 细胞因子如IP-10和TNF α。此外，我们提出，毛地黄黄酮降低血压， 抑制TNF-α和sFlt 1诱导的内皮细胞产生ET-1。为了验证这个假设，我们将使用 人胎盘外植体、细胞滋养层细胞系、培养的内皮细胞、分离的血管，以及 作为新型PE动物模型中的分子、药理学和生理学技术， 子宫灌注压（RUPP）模型和sFlt 1模型。我们的具体目标和中心假设是 有着强大的初步和可行性数据支持。拟议的实验将确定机制， 毛地黄黄酮干扰胎盘缺血与母体心血管疾病的病理生理通路 功能障碍，同时产生临床前数据，以支持木樨草素作为治疗PE的发展。

PROJECT SUMMARY/ABSTRACT Preeclampsia (PE) is a common and life-threatening hypertensive disorder of pregnancy. This disease affects nearly 7% of all pregnancies and is a leading cause of maternal and fetal morbidity and mortality. There are currently no therapies that treat PE except for early delivery. Data from our lab and others suggest that women with PE have angiogenic imbalance (excess soluble fms-like tyrosine kinase 1 [sFlt1]), evidence of inflammation (excess tumor necrosis factors TNF-α, interleukins such as IFN-𝛾-inducible protein 10-IP-10 and IL-6) and excess reactive oxygen species (ROS). The release of these soluble factors from the placenta into the bloodstream interferes with the supply of pro-angiogenic factors, such as placental growth factor (PlGF), resulting in reduced nitric oxide (NO) and increased endothelin-1 (ET-1), and ultimately endothelial dysfunction and hypertension. The lack of treatment options for PE likely results from its complex etiology and limitations on drug use during pregnancy. Identifying a safe therapeutic for this condition is a major unmet need for the health of pregnant women. The availability of chemical libraries and high-throughput screening methodologies has allowed investigators to discover novel molecules that interfere with signaling pathways involved in disease biology. We screened a chemical library of natural compounds and identified the bioflavonoid, luteolin, as a potent inhibitor of sFlt1 in human placental cell lines and placental explants. In addition, we discovered that luteolin inhibits a variety of pro-inflammatory cytokines (such as IL-1, IL-27, IL-6 and IP-10) and interrupts distinct cellular pathways that inhibit several key proteins implicated in pathogenesis of PE. Specifically, we found the luteolin is a potent inhibitor of hypoxia-inducible factor 1α (HIF-1α, a major regulator of sFlt1). In addition, luteolin reduces TNF- α induced ET-1 production in vitro, reduces blood pressure, relaxes uterine arteries, improves placental perfusion in placental ischemia rat models of PE. Our exciting and novel preliminary data led us to propose the central hypothesis that luteolin improves uterine perfusion and reduces total peripheral resistance and blood pressure in models of PE by reducing placental production of sFlt1 and HIF-1α and inhibiting cytokines such as IP-10 and TNF α. In addition, we propose that luteolin decreases blood pressure by inhibiting TNF-α and sFlt1-induced endothelial production of ET-1. To test this hypothesis, we will use human placental explants, cytotrophoblast cell lines, cultured endothelial cells, isolated blood vessels, as well as molecular, pharmacological and physiological techniques in novel animal models of PE, the Reduction in Uterine Perfusion Pressure (RUPP) model and the sFlt1 model. Our specific aims and central hypothesis are backed by strong preliminary and feasibility data. The proposed experiments will identify mechanisms by which luteolin interferes with the pathophysiological pathways linking placental ischemia with maternal cardiovascular dysfunction, while generating preclinical data to support the development of luteolin as a therapy for PE.