ENDOCRINE & PARACRINE RELATIONSHIPS IN PARTURITION

这项研究计划的目标是澄清 内分泌、旁分泌和细胞事件之间的关系 在灵长类动物中达到分娩的顶峰。怀孕期间获得的数据 恒河猴将提供对正常和 人类早产。促肾上腺皮质激素的生理学研究 促肾上腺皮质激素(ACTH)、促皮质激素释放激素(CRH)、雄激素、 糖皮质激素和白介素8输注联合阻断 雄激素、雌激素和黄体酮的作用将在 长期插管、活动、怀孕的恒河猴。 补充子宫内组织的体外研究将解决 以下问题：(1)恒河猴的过早激活 胎儿(或母亲)下丘脑-垂体-肾上腺(HPA)轴 复制自发的细胞和旁分泌事件 分娩；会导致早产和分娩吗？(2) 雄激素、雌激素对胎儿血浆ACTH升高的影响 或作用于羊膜/绒毛或蜕膜的糖皮质激素？(3)有吗？ 宫内“孕酮(P)撤退”发生在子宫肌层或 蜕膜P受体亚型(PR-A和PR-A)的妊娠变化 PR-B)或羊膜、免疫细胞或子宫肌细胞中的非基因组 机制(即，膜效应)？(4)直接羊膜腔内 输注纯化的IL-8复制趋化/旁分泌 自然分娩或ACTH诱导分娩的表现？子宫 将持续监测、量化和系列化收缩能力 将对羊水、母体和胎儿血液样本进行检测 对于类固醇激素，促肾上腺皮质激素、促肾上腺皮质激素释放激素、催产素、催乳素、二十烷醇， 细胞因子和热休克蛋白。宫内组织标本 将在分娩或剖腹产时获得组织病理学 研究；细胞因子、性类固醇和 糖皮质激素受体和前列腺素脱氢酶； 信使核糖核酸的定量(细胞因子、性激素受体，3 -羟基类固醇脱氢酶和环氧合酶亚型)和 基质金属蛋白酶的酶谱检测。羊水中的白细胞 液体将通过流式细胞术进行评估；其潜在的胎儿来源 通过扩增Y染色体特异DNA。隔离状态下的钙离子通量 羊膜、免疫细胞和子宫肌细胞将通过 荧光成像。对于在线数据，将使用计算机方法 采集和统计分析波形、生物节律和 荷尔蒙趋势。通过比较和整合生理学， 试验组和对照组的激素和细胞变化，我们 将获得对建立因果关系至关重要的证据 负责灵长类动物的足月和早产。

The objectives of this research plan are to clarify the relationships among endocrine, paracrine and cellular events which culminate in parturition in primates. Data obtained in pregnant rhesus monkeys will provide insights into the mechanisms of normal and preterm human labor. Physiological studies of adrenocorticotropin (ACTH), corticotropin releasing hormone (CRH), androgen, glucocorticoid and interleukin (IL)-8 infusions together with blockade of androgen, estrogen and progesterone action will be carried out in chronically-catheterized, mobile, pregnant rhesus monkeys. Complementary in vitro studies of intrauterine tissues will address the following questions (1) Does premature activation of the rhesus fetal (or maternal) hypothalamic-pituitary-adrenal (HPA) axis replicate the cellular and paracrine events seen in spontaneous parturition; does it lead to preterm labor and delivery? (2) Are the effects of rising fetal plasma ACTH mediated by androgens, estrogens or glucocorticoids acting upon amnion/chorion or decidua? (3) Does intrauterine "progesterone (P) withdrawal" occur in myometrium or decidua by a gestational change in the P receptor isoforms (PR-A and PR-B) or in amnion, immune cells or uterine myocytes by nongenomic mechanisms (i.e., membrane effects)? (4) Does direct intraamniotic infusion of purified IL-8 replicate the chemotactic/paracrine manifestations of spontaneous or ACTH-induced parturition? Uterine contractility will be continuously monitored and quantified and serial samples of amniotic fluid, maternal and fetal blood will be assayed for steroid hormones, ACTH, CRH, oxytocin, prolactin, eicosanoids, cytokines, and heat shock proteins. Samples of intrauterine tissues will be obtained at delivery or cesarean section for histopathologic studies; immunohistochemistry for cytokines, sex steroid and glucocorticoid receptors and prostaglandin dehydrogenase (PGDH); quantification of mRNA (for cytokines, sex steroid receptors, 3 -hydroxysteroid dehydrogenase and cyclooxygenase isoforms) and zymography for matrix metalloproteinases. Leukocytes in amniotic fluid will be assessed by flow cytometry; their potential fetal source by amplification of Y-chromosome specific DNA. Ca2+ flux in isolated amnion, immune cells and uterine myocytes will be measured by fluorescent imaging. Computer methods will be used for on-line data acquisition and statistical analyses of waveforms, biorhythms, and hormonal trends. By comparing and integrating the physiologic, hormonal and cellular changes in experimental groups and controls, we will obtain evidence which is critical to establish the causal links responsible for term and preterm labor in a primate.