Bioassay Core/ Judy L. Bolton

生物测定核心/ Judy L. Bolton

• 批准号: 8007079
• 负责人: NORMAN R FARNSWORTH
• 金额: $ 18.09万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-30 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8007079
• 关键词: Affinity; Alkaline Phosphatase; Anti-Inflammatory Agents; Anti-inflammatory; Antioxidants; Aromatase; Aromatase Inhibition; Autopsy; Binding; Biological; Biological Assay; Biological Markers; Biological Process; Blood; Botanicals; Breast Cancer Cell; Cancer cell line; Cell Line; Cells; Chemoprevention; Chemopreventive Agent; Clinical; Collaborations; Competitive Binding; Cyclic AMP; Cytochrome P450; Data; Development; Drug Metabolic Detoxication; Endometrial; Endometrium; Enzymes; Estrogen Antagonists; Estrogen Metabolism; Estrogen Receptors; Estrogen receptor negative; Estrogens; Evaluation; Fractionation; Future; Glutathione S-Transferase; Goals; Grant; Hormonal; Hot flushes; Lead; Ligands; Liver; Luciferases; Mammary gland; Menopausal Symptom; Metabolism; Modeling; Modification; NAD(P)H dehydrogenase (quinone) 1, human; Organ; Pharmaceutical Preparations; Phase; Plant Extracts; Poison; Postmenopause; Process; Progesterone; Progesterone Receptors; Proliferating; Property; Radioactive; Rattus; Recommendation; Research; Response Elements; Role; Safety; Sampling; Screening procedure; Selective Serotonin Reuptake Inhibitor; Serotonin; Standardization; T47D; Testing; Tissues; Toxic effect; Ultrafiltration; Women' s Health; Work; Xenobiotics; base; carcinogenesis; chemical standardization; cost effective; cytotoxic; cytotoxicity; dietary supplements; enzyme activity; high throughput screening; improved; in vivo; receptor binding; research study; reuptake

The new Bioassay Core (Core C) will provide bioassay support for all Projects with an array of validated, robust, and efficient bioassays, which have been previously established in Project 2. The following specific aims are proposed: 1. Evaluate botanical extracts for their chemopreventive properties. We will analyze the antioxidative activity, the NAD(P)H:quinone oxidoreductase 1 (NQ01) inducing properties, the antiinflammatory activity, and aromatase enzyme inhibiting properties of the extracts/compounds. When extracts/compounds will be active in the NQ01 assay, we will analyze these samples in the antioxidative response element luciferase assay and in the glutathione-S-transferase induction assay. The induction of several Phase I metabolism enzymes is regulated by the xenobiotic response element (XRE). Inducer of Phase I enzymes interfere with the metabolism of other drugs and endogenous estrogen; therefore, the extracts will be tested in the XRE-luciferase assay. 2. Evaluate botanical extracts/compounds for hormonal activity (efficacy). The alkaline phosphatase assay in Ishikawa cells will be employed as the major assay to test the botanicals for estrogenic and antiestrogenic activities. Active fractions will be analyzed for estrogen receptor (ER)a and ERB selectivity in an ERa and ERB-ERE-luciferase assay in breast cancer cells. If promising samples have been found, we will test for ER ligands in the ERa and ERB competitive radioactive binding assay. As progestogenic activity is important for endometrium protection, the progesterone response element luciferase and progesterone receptor competitive binding assay will be conducted. Finally, the selective serotonin reuptake inhibition assay (SSRI) will be employed, since SSRIs have been described to alleviate menopausal symptoms. 3. Evaluate botanicals for their distribution profile, safety, and efficacy in vivo. When active extracts/compounds have been determined, the ovariectomized rat model to analyze the distribution and metabolism profile (Project 3), the safety, and efficacy will be performed. With these assays. Core C directly supports the whole Center efforts by establishing toxic, interfering, and active compounds, thus providing bioactive marker compounds for the standardization of new botanicals.

新的生物测定核心（核心 C）将为所有项目提供生物测定支持，并提供一系列经过验证的、 稳健、高效的生物测定，这些已在项目 2 中建立。以下具体内容 提出的目标是： 1. 评估植物提取物的化学预防特性。我们将分析 抗氧化活性、NAD(P)H:醌氧化还原酶 1 (NQ01) 诱导特性、抗炎 提取物/化合物的活性和芳香酶抑制特性。什么时候 提取物/化合物将在 NQ01 测定中具有活性，我们将在抗氧化中分析这些样品 反应元件荧光素酶测定和谷胱甘肽-S-转移酶诱导测定。的归纳 多种 I 期代谢酶受异生素反应元件 (XRE) 调节。诱导剂 I相酶干扰其他药物和内源性雌激素的代谢；因此， 提取物将在 XRE 荧光素酶测定中进行测试。 2. 评估植物提取物/化合物的激素水平 活性（功效）。石川细胞中的碱性磷酸酶测定将作为主要测定方法 测试植物药的雌激素和抗雌激素活性。将分析活性部分的雌激素 乳腺癌细胞中 ERa 和 ERB-ERE-荧光素酶测定中的受体 (ER)a 和 ERB 选择性。如果 已经发现有希望的样品，我们将测试ERa和ERB竞争性放射性中的ER配体 结合测定。由于孕激素活性对于子宫内膜保护很重要，因此孕激素 将进行反应元件荧光素酶与孕激素受体竞争性结合测定。 最后，将采用选择性血清素再摄取抑制测定（SSRI），因为 SSRI 已被 据说可以缓解更年期症状。 3. 评估植物药的分布情况、安全性和 体内功效。当确定活性提取物/化合物后，切除卵巢的大鼠模型 分析分布和代谢概况（项目 3）、安全性和有效性。和 这些测定。核心 C 通过建立有毒、干扰和活性物质直接支持整个中心的工作 化合物，从而为新植物药的标准化提供生物活性标记化合物。