IN VITRO STUDIES OF INFECTION-MEDIATED PRETERM LABOR

感染介导的早产的体外研究

• 批准号: 2201026
• 负责人: DOUGLAS A KNISS
• 金额: $ 8.94万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-02-01 至 1999-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2201026
• 关键词: amnion; arachidonate; biological signal transduction; cytokine; enzyme activity; epidermal growth factor; growth factor receptors; human pregnant subject; interleukin 1; medical complication; messenger RNA; northern blottings; nucleic acid hybridization; nucleic acid probes; phospholipase A2; pregnancy infection; premature labor; prostaglandin endoperoxide synthase; receptor expression; tissue /cell culture; tumor necrosis factor alpha; tumor necrosis factor beta; western blottings; women's health

Clinical studies suggest that intrauterine infection is a major medical factor association with preterm labor (PTL) in women. We hypothesize that PTL results from the synergistic interplay between signals (e.g. EGF) and inflammatory cytokines (e.g. IL-I, TNF-alpha, TGF-beta), elaborated into the decidua and/or amniotic cavity following a maternal immune response to bacterial colonization. We postulate that cytokines modulate the action of EGF by sensitizing amnion cells and, thereby, cause a robust prostaglandin synthetic response leading to premature uterine contractility and cervical dilatation. This research proposal will test the hypothesis that cytokines modulate EGF action in amnion cells using the amnion-derived cell line WISH as an in vitro model system. The EGF signal transduction cascade (i.e. receptor expression, phospholipase A2-mediated arachidonic acid release, and cyclooxygenase activation/induction) will be examined to determine whether cytokine treatment alters EGF action at one or more of these key regulatory points. Aim 1 will address whether IL-1beta, TNF- alpha and TGF-beta, which are associated with intrauterine infection- mediated PTL can alter EGF receptor expression by (i) inducing rapid EGF receptor internalization and recycling to the cell surface and (ii) increasing the expression of EGF receptor mRNA in amnion cells. Aim 2 will examine whether IL-1beta, TNF-alpha and TGF-beta can modulate phospholipase A2 activity and/or induction either independently or in combination with EGF in amnion cells. This question will be addressed by measuring the specific activity, PLA2 protein expression and PLA2 mRNA expression in cytokine and EGF treated cells. Aim 3 will examine whether IL-1beta, TNF-alpha and TGF-beta can modulate cyclooxygenase activity and/or induction either independently or in combination with EGF in amnion cells. The specific activity for cyclooxygenase, and cyclooxygenase protein and mRNA expression will be measured to address this aim. Elucidating the potential control points for cytokine-mediated alterations in EGF action in amnion cells may provide a readily testable model system for understanding the biochemical details of premature activation of the prostaglandin cascade in relation to PTL.

临床研究表明宫内感染是主要的医学 妇女中与早产（PTL）的因素关联。我们假设这一点 PTL来自信号之间的协同相互作用（例如EGF）和 炎症细胞因子（例如IL-I，TNF-Alpha，TGF-Beta），详细阐述 母体免疫反应后的deciDUA和/或羊水腔 细菌定植。我们假设细胞因子调节 例如，通过使羊膜细胞敏化，从而引起强大的前列腺素 合成反应导致子宫收缩性过早和宫颈 扩张。该研究建议将检验细胞因子的假设 使用羊膜衍生的细胞系在羊膜细胞中调节EGF作用 希望作为体外模型系统。 EGF信号转导级联 （即受体表达，磷脂酶A2介导的花生四烯酸 将检查释放和环氧合酶激活/诱导） 确定细胞因子治疗是否改变了一个或多个的EGF作用 这些关键的监管点。 AIM 1将解决IL-1Beta，tnf-是否 α和TGF-β，与宫内感染有关 介导的PTL可以通过（i）诱导快速EGF改变EGF受体表达 受体内在化并回收到细胞表面，（ii） 增加羊膜细胞中EGF受体mRNA的表达。 AIM 2意志 检查IL-1BETA，TNF-ALPHA和TGF-BETA是否可以调节 磷脂酶A2活性和/或诱导独立或诱导 与羊膜细胞中的EGF结合。这个问题将由 测量特定活性，PLA2蛋白表达和PLA2 mRNA 在细胞因子和EGF处理的细胞中的表达。 AIM 3将检查是否 IL-1BETA，TNF-ALPHA和TGF-BETA可以调节环氧合酶活性 和/或独立诱导或与羊膜中的EGF结合 细胞。环氧酶和环氧合酶的特定活性 将测量蛋白质和mRNA表达来解决这一目标。 阐明细胞因子介导的改变的潜在控制点 在羊膜细胞中的EGF作用中，可能会提供一个易于测试的模型系统 了解过早激活的生化细节 与PTL有关的前列腺素级联。