ETHANOL AND PLACENTAL TOXICITY SIGNAL CROSS TALK

乙醇和胎盘毒性信号的交互作用

• 批准号: 6168218
• 负责人: STANLEY E. FISHER
• 金额: $ 34.94万
• 依托单位: SUNY DOWNSTATE MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-02-01 至 2002-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6168218
• 关键词: G protein; adenylate cyclase; biological signal transduction; cyclic AMP; diacylglycerols; embryo /fetus toxicology; enzyme activity; ethanol; fetal alcohol syndrome; human tissue; ligands; phosphorylation; placental transfer; protein isoforms; protein kinase C; receptor binding; receptor expression; tissue /cell culture; trophoblast

The placenta is a multifunctional organ of fetal origin, which is critical to normal fetal growth and development. In addition to direct fetal toxicity, ethanol may be toxic to the placenta as well. Ethanol-induced placental toxicity could contribute to the pathophysiology of alcohol related fetal injury. During the course of the current grant, this laboratory has characterized human placental trophoblasts in culture and demonstrated several alterations in cellular physiology. The human trophoblast is the principal functional cell of the human placenta, but it also expresses physiologic properties which are found in other human cell types. The ethanol-treated, cultured trophoblast model system, developed in this laboratory, enables evaluation of molecular biochemistry in readily available, nontransformed human cells. Using this system, we propose to test the fundamental hypothesis: Ethanol alters intracellular signal transduction. In particular, we will concentrate on the effect of ethanol on signal "cross-talk" mediated by protein kinase C (PKC), an important regulatory factor in normal intracellular signal transduction. This laboratory has shown that chronic exposure to ethanol, within a dose range found in man, results in enhancement of ligand-stimulated cAMP production by cultured trophoblasts. Preliminary data indicate that this alteration is not due to quantitative changes in G protein expression. Rather, there is an effective increase in adenylyl cyclase (AC) activity. We propose that this may be due, at least in part, to ethanol-induced activation of PKC and subsequent interaction of PKC with components of the adenylyl cyclase system. This proposal will investigate the biochemical basis for ethanol-induced enhancement of ligand-stimulated cAMP production in the context of PKC mediated signal "cross talk." Using the general model system of the cultured human placental trophoblast, exposed to ethanol in vitro, we propose to: Investigate ethanol-induced changes in components of the AC signalling system which are modulated by PKC "cross- talk;" Use inhibition of PKC to evaluate the role of PKC in ethanol- induced changes in the AC system; Evaluate the effect of ethanol on PKC activity, as it relates to the several phospholipases and diacylglycerol production; Determine the role of phosphatidylethanol as an activator of PKC and its role in signal "cross-talk." The studies should provide new information on the cellular mechanisms by which ethanol alters placental function and, hence, advance our understanding of the pathophysiology of the Fetal Alcohol Syndrome. Additionally, these findings should contribute to the general understanding of the mechanisms by which ethanol affects the biology of many human tissues, including those of the fetus.

胎盘是胎儿起源的多功能器官， 正常的胎儿生长发育 除了直接胎儿 毒性，乙醇也可能对胎盘有毒。 乙醇诱导 胎盘毒性可能有助于酒精的病理生理学 相关的胎儿损伤 在目前的拨款过程中，这 一个实验室已经在培养中表征了人胎盘滋养层细胞， 在细胞生理学上表现出了一些变化。 人类 滋养层是人类胎盘的主要功能细胞，但它 也表达在其他人类细胞中发现的生理特性 类型 乙醇处理，培养的滋养层模型系统，开发 在这个实验室，使分子生物化学的评价， 容易获得的未转化的人类细胞。 利用这个系统，我们 我建议测试基本假设：乙醇改变细胞内 信号转导 特别是，我们将集中讨论 乙醇对蛋白激酶C（PKC）介导的信号“串扰”的影响， 在正常细胞内信号转导中重要调节因子。 这个实验室已经证明，长期接触乙醇，在一定剂量内， 在人类中发现的范围，导致配体刺激的cAMP增强 由培养的滋养层细胞产生。 初步数据显示， 改变不是由于G蛋白表达的定量变化。 相反，腺苷酸环化酶（AC）活性有效增加。 我们认为，这可能是由于，至少部分是由于乙醇诱导的 PKC的激活以及随后PKC与 腺苷酸环化酶系统。 这项提案将研究生物化学 乙醇诱导的配体刺激的cAMP产生增强的基础 在PKC介导的信号“串扰”的背景下。“利用一般 培养的人胎盘滋养层细胞的模型系统，暴露于 乙醇在体外，我们建议：研究乙醇诱导的变化， AC信号系统的组分，其由PKC“交叉- 讨论;”使用PKC的抑制来评估PKC在乙醇中的作用- AC系统的诱导变化;评价乙醇对PKC的影响 活性，因为它涉及几种磷脂酶和甘油二酯 生产;确定磷脂酰乙醇作为活化剂的作用， PKC及其在信号“串扰”中的作用。“这些研究应该提供新的 乙醇改变胎盘细胞机制的信息 功能，因此，推进我们的病理生理学的理解， 胎儿酒精综合症 此外，这些发现应 有助于对乙醇的作用机制的普遍理解， 影响许多人体组织的生物学，包括胎儿的组织。

项目成果

Ethanol enhancement of ligand-stimulated cAMP production by cultured human placental trophoblasts.

乙醇增强培养的人胎盘滋养层配体刺激的 cAMP 产生。

• DOI: 10.1016/0006-2952(94)90575-4
• 发表时间: 1994
• 期刊: Biochemical pharmacology
• 影响因子: 5.8
• 作者: Karl,PI;Divald,A;Fisher,SE
• 通讯作者: Fisher,SE

Altered cyclic AMP-dependent human chorionic gonadotropin production in cultured human placental trophoblasts exposed to ethanol.

暴露于乙醇的培养人胎盘滋养层中环磷酸腺苷依赖性人绒毛膜促性腺激素的产生发生改变。

• DOI: 10.1016/s0006-2952(97)00404-8
• 发表时间: 1998
• 期刊: Biochemical pharmacology
• 影响因子: 5.8
• 作者: Karl,PI;Divald,A;Diehl,AM;Fisher,SE
• 通讯作者: Fisher,SE

Chronic ethanol exposure inhibits insulin and IGF-1 stimulated amino acid uptake in cultured human placental trophoblasts.

慢性乙醇暴露会抑制培养的人胎盘滋养层中胰岛素和 IGF-1 刺激的氨基酸摄取。

• DOI: 10.1111/j.1530-0277.1994.tb00063.x
• 发表时间: 1994
• 期刊: Alcoholism, clinical and experimental research
• 作者: Karl,PI;Fisher,SE
• 通讯作者: Fisher,SE

Biotin transport in microvillous membrane vesicles, cultured trophoblasts, and isolated perfused human placenta.

微绒毛膜囊泡、培养的滋养层细胞和分离的灌注人胎盘中的生物素转运。

• DOI: 10.1152/ajpcell.1992.262.2.c302
• 发表时间: 1992
• 期刊: The American journal of physiology
• 作者: Karl,PI;Fisher,SE
• 通讯作者: Fisher,SE

Na(+)-dependent amino acid uptake by human placental microvillous membrane vesicles: importance of storage conditions and preservation of cytoskeletal elements.

人胎盘微绒毛膜囊泡对Na()依赖性氨基酸的摄取：储存条件和细胞骨架元素保存的重要性。

• DOI: 10.1016/0143-4004(91)90005-z
• 发表时间: 1991
• 期刊: Placenta
• 影响因子: 3.8
• 作者: Karl,PI;Teichberg,S;Fisher,SE
• 通讯作者: Fisher,SE