Benefits of nicotinamide in placental development and in preeclamsia

摘要/摘要 子痫前期(PE)发病的分子机制目前尚不清楚，有效的预防措施 治疗策略仍然难以捉摸。PE是一种妊娠高血压疾病，并伴有 在美国，大约每20个怀孕中就有一个是与怀孕相关的孕产妇死亡的主要原因 以及全世界的新生儿发病率/死亡率。内皮素-1(ET-1)是一种由21个残基组成的缩血管多肽， 而编码ET-1前体的EDN1基因的单核苷酸多态(SNPs)与PE相关。 Edn1H/Edn1表达上调至正常3倍的小鼠，尽管血压升高，但血压正常 循环中的ET-1。然而，Edn1H/DAMs在怀孕后期发育出全谱的PE样表型。 此外，来自Edn1H/DAMs的胚胎，无论其Edn1基因类型如何，在 着床早期，外胎盘锥体方向混乱。我们报告了烟酰胺(酰胺形式的 ADP核糖环化酶下游的ET-1抑制剂维生素B3(NAM)改善脑缺血再灌注损伤的PE样表型 两种不同的PE小鼠模型，我们的初步数据显示，NAM可降低尿白蛋白 排泄，并增加存活的胎儿数量时，在整个治疗期间 怀孕了。这些观察结果使我们假设Edn1H/Dams中的PE起源于异常 植入早期母体ET-1高表达所致胎盘形成，NAM可纠正 这会破坏和保护大坝，使其不受后期PE开发的影响。因此，特定目标1将测试这一点 分离NAM对Edn1H/大坝PE的早期影响和后期影响的假说 维生素从不同的妊娠阶段开始，持续不同的时间。妊娠结局，包括血液 在性交后18.5天(DPC)测定血压、尿白蛋白、胎儿数量和体重。在……里面 此外，植入阶段ET-1系统各组成部分的表达以及NAM对它们的影响将 接受检查。特异性靶点2将探讨ET-1和NAM对分化的影响机制 二维和三维培养从人滋养层干细胞分化为指定滋养层细胞 系统。药理剂量的ET-1单独，或ET-1加NAM将添加到特定的条件 5~6成熟。将通过细胞的形态、运动性和不同类型的标记的表达来检查细胞 滋养层细胞。特异靶3将探讨ET-1和NAM对受损子宫的作用机制 蜕膜化和血管生成。子宫内膜造口分化的标志物--血液结构 胚胎着床期子宫血管及血管内皮生长因子的表达 将对怀孕进行检查。原代培养的子宫内膜间质细胞将被药物处理 单独应用ET-1，或ET-1加NAM，分化标志物和血管内皮生长因子的表达 下定决心。拟议的研究将扩大和加深我们对 母体遗传因子ET-1对PE的影响，并确定潜在的PE干预策略。

Summary/Abstract The molecular mechanism of pathogenesis of preeclampsia (PE) is largely unknown, and effective prevention and treatment strategies remain elusive. PE is a pregnancy-associated hypertensive condition and complicates approximately one in 20 pregnancies in the US and is a leading cause of pregnancy-related maternal mortality and neonatal morbidity/mortality worldwide. Endothelin-1 (ET-1) is a vasoconstrictive peptide of 21 residues, and single nucleotide polymorphisms (SNPs) in EDN1, coding for a precursor for ET-1, are associated with PE. Edn1H/+ mice in which Edn1 expression is elevated to 3X normal have normal blood pressure, despite elevated circulating ET-1. However, Edn1H/+ dams develop full spectrum of PE-like phenotypes in their late pregnancy. In addition, the embryos from Edn1H/+ dams, regardless of their Edn1 genotypes, lag in development during early implantation stage with disoriented ectoplacental cones. We reported that nicotinamide (amide form of vitamin B3, Nam), inhibitor of ET-1 downstream of ADP ribosylcyclase, ameliorates the PE-like phenotypes in two separate mouse models of PE, and our preliminary data show that Nam decreases urinary albumin excretion and increases the number of survival fetuses when Edn1H/+ dams were treated during the entire pregnancy. These observations have led us to hypothesize that PE in Edn1H/+ dams originates from abnormal placentation caused by a high maternal ET-1 expression at early implantation stage, and that Nam can correct this damage and protect dams from later PE development. Accordingly, Specific Aim 1 will test this hypothesis by dissociating early effects from later effects of Nam on PE of Edn1H/+dams by treatments with this vitamin starting at different gestational stages and for different durations. Pregnancy outcomes including blood pressure, urinary albumin and fetal number and weight will be determined at 18.5 day post coitus (dpc). In addition, the expression of components of ET-1 system and Nam’s effects on them at implantation stage will be examined. Specific Aim 2 will investigate the mechanism of effects of ET-1 and Nam on differentiation from human trophoblast stem cells into designated trophoblast cells by using 2- and 3- dimension culture system. Pharmacological dose of ET-1 alone, or ET-1 plus Nam will be added to the specific conditioned medium. Cells will be examined by their morphology, motility, and expression of markers of different types of trophoblasts. Specific Aim 3 will investigate the mechanism of effects of ET-1 and Nam on impaired uterine decidualization and angiogenesis. The markers of endometrial stomal differentiation, the structure of blood vessels and the expression of vascular endothelial growth factor (VEGF) in uteri at the implantation stage of pregnancy will be examined. Primary cultured endometrial stromal cells will be treated with pharmacological dose of ET-1 alone, or ET-1 plus Nam, and the markers of differentiation and the expression of VEGF will be determined. The proposed research will broaden and deepen our understanding regarding the role of the maternal genetic factor ET-1 on PE and identify a potential intervention strategy for PE.