MECHANISMS OF ACTION IN MENOPAUSE

更年期的作用机制

• 批准号: 6954972
• 负责人: Judy L Bolton
• 金额: $ 20.45万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2010-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6954972
• 关键词: alternative medicine; antioxidants; bioassay; biological models; chemoprevention; chromatography; depression; dietary supplements; drug screening /evaluation; emotions; female; hormone regulation /control mechanism; hot flash; laboratory rat; medicinal plants; menopause; nutrition related tag; pharmacokinetics; phytoestrogens; plant extracts; serotonin; serotonin receptor; women' s health

The major objective of Project by Bolton continues to be to provide in vitro bioassay support for the discovery of the active components for the major dietary supplements for the relief of menopausal symptoms. To achieve this objective several in vitro cell based and animal assays have been developed in order to investigate the mechanism of action of these dietary supplements. Specifically, we are interested in botanicals with estrogenic activity such as hops (Humulus lupulus), kudzu (Pueraria lobata), and kwao keur (Pueraria mirifica). In addition, we have discovered an alternative serotonergic mechanism for the relief of menopausal symptoms which could be important for black cohosh (Cimicifuga racemosa), dong quai (Angelica sinensis), and valerian (Valeriana officinalis). The hypothesis to be addressed is whether select plant extracts contain potent estrogenic and/or serotonergic compounds useful for the relief of menopausal symptoms as well as other beneficial properties of estrogens such as antioxidant/chemopreventive effects. The following specific aims are proposed: 1. To evaluate the hormonal activity of plant extracts and active compounds. We have already screened a number of plant extracts and pure compounds for estrogenic effects and investigated their mechanism of action using a variety of in vitro assays including competitive ER binding assays, induction of estrogen sensitive genes, and ERE-luciferase assays. In addition, we have tested several of these extracts for hormonal activity in vivo using the ovariectomized rat model. We have also incorporated two new screening assays including the estrogen receptor coactivator assay and an assay to measure ERE and AP-1 DNA binding capacity. 2. To evaluate the serotonergic activity of plant extracts and active compounds. We plan to further fractionate black cohosh (aim 3) in order to determine the active compounds as well as test other promising plants using established serotonergic assays (competitive binding to serotonin receptors, elevation in cyclic AMP activity, and SSRI assays), addition, we plan to further test the hypothesis that the 5-HT7 receptor is involved in thermoregulation (i.e. hot flashes) whereas the 5-HT1A and 5-HT5A receptors are likely responsible for depression and mood. Animal models will be incorporated including the hot flash model and the forced swim test (mood). 3. To evaluate the chemoprevention/antioxidant properties of plant extracts and active compounds. During the previous granting period, we have developed a variety of antioxidant/chemoprevention assays including scavenging of stable free radicals, inhibition of DNA damage induced by the quinone menadione using the comet assay, and induction of quinone reductase. In this aim, extracts of the above mentioned botanical dietary supplements will be fractionated and activity will be determined using the antioxidant/chemoprevention assays. The completion of these aims will determine which specific botanicals have health benefits for relief of menopausal symptoms and determine their mechanism of action.

博尔顿项目的主要目标仍然是为发现缓解更年期症状的主要膳食补充剂的活性成分提供体外生物测定支持。为了实现这一目标，已经发展了几种基于细胞和动物的体外试验，以研究这些饮食的作用机制。 补充剂。具体地说，我们对具有雌激素活性的植物感兴趣，如啤酒花(Humulus Lupulus)、葛根(Grouaria Lobata)和葛根(Kwao Keur)。此外，我们还发现了一种替代的5-羟色胺能机制来缓解更年期症状，这对黑升麻(Cimicifuga Racemosa)、冬瓜(Angelica Sinensis)和缬草(Valeriana Office Inalis)可能是重要的。要解决的假设是，选定的植物提取物是否含有有效的雌激素和/或5-羟色胺能化合物，有助于缓解更年期症状，以及雌激素的其他有益特性，如抗氧化/化学预防作用。具体目标如下：1.评价植物提取物和活性化合物的激素活性。我们已经放映了一部 对植物提取物和纯化合物的雌激素样作用进行了研究，并利用多种体外试验，包括竞争性ER结合试验、雌激素敏感基因的诱导和Ere-荧光素酶试验，研究了它们的作用机制。此外，我们使用去卵巢的大鼠模型，在体内测试了其中几种提取物的激素活性。我们还加入了两种新的筛选方法，包括雌激素受体共激活物方法和一种测定ERE和AP-1 DNA结合能力的方法。2.评价植物提取物和活性化合物的5-羟色胺能活性。我们计划进一步分离黑升麻(目标3)，以确定活性化合物，并使用已建立的5-羟色胺能分析法(与5-羟色胺受体的竞争结合、环磷酸腺苷活性升高和SSRI分析)测试其他有希望的植物，此外，我们计划进一步测试 假设5-HT7受体参与体温调节(即潮热)，而5-HT1A和5-HT5A受体可能与抑郁和情绪有关。动物模型将被纳入，包括潮热模型和强迫游泳测试(情绪)。3.评价植物提取物和活性化合物的化学预防/抗氧化性能。在之前的授予期间，我们开发了各种抗氧化剂/化学预防方法，包括清除稳定的自由基，使用彗星试验抑制苯二酮诱导的DNA损伤，以及诱导苯醌还原酶。为此，上述植物膳食补充剂的提取物将被分级，并将使用抗氧化剂/化学预防试验来确定活性。这些目标的完成将决定哪些特定的植物具有健康 有助于缓解更年期症状，并确定其作用机制。