Bioassay Core/ Judy L. Bolton

新的生物检测核心(核心C)将为所有项目提供生物检测支持， 健壮、高效的生物检测方法，这是以前在项目2中建立的。 目的是：1.评价植物提取物的化学预防性能。我们将分析 抗氧化活性，NAD(P)H：苯醌氧化还原酶1(NQ01)的诱导特性，抗炎 提取物/化合物的活性和芳香酶抑制性能。什么时候 提取物/化合物将在NQ01检测中发挥作用，我们将分析这些样品中的抗氧化剂 反应元件荧光素酶测定和谷胱甘肽-S-转移酶诱导试验。的归纳 几种I相代谢酶受异种生物反应元件(XRE)的调节。诱导者 I相酶干扰其他药物和内源性雌激素的新陈代谢；因此， 提取物将在XRE-荧光素酶分析中进行测试。2.评估植物提取物/化合物的荷尔蒙 活性(功效)。Ishikawa细胞碱性磷酸酶检测将作为主要检测方法 测试植物类雌激素和抗雌激素活性。活性部分将被分析雌激素 ERA和ERB在乳腺癌细胞中的选择性和ERB-ERE-荧光素酶分析。如果 有希望的样本已经找到，我们将检测ER配体在时代和ERB的竞争性放射性 结合试验。由于孕激素活性对子宫内膜的保护很重要，所以孕酮 将进行反应元件荧光素酶和孕激素受体竞争结合试验。 最后，将采用选择性5-羟色胺再摄取抑制试验(SSRI)，因为SSRI已经 被描述为缓解更年期症状。3.评估植物性药物的分布概况、安全性和 体内药效。当活性提取物/化合物被确定后，去卵巢的大鼠模型 分析药物的分布和代谢特征(项目3)，进行安全性和有效性分析。使用 这些化验结果。核心C通过建立有毒、干扰和主动的机制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。

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.