https://scholars.lib.ntu.edu.tw/handle/123456789/414032
標題: | Establishment of reporter platforms capable of detecting NF-£eB mediated immuno-modulatory activity | 作者: | Wang C.-H. Lin J.-H. Lu T.-J. Chiang A.-N. Chiou S.-T. Chen Y.-A. Pan M.-H. SHU-CHEN HSIEH MIN-HSIUNG PAN |
關鍵字: | cell platform;COX-2;iNOS;luciferase assay;RAW264.7 | 公開日期: | 2013 | 卷: | 61 | 期: | 51 | 起(迄)頁: | 12582-12587 | 來源出版物: | Journal of Agricultural and Food Chemistry | 摘要: | Cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) are transcriptional targets of nuclear factor kappa B (NF-£eB) that are involved in inflammatory responses. The aim of this study is to develop a method for efficiently detecting inflammation modulatory activities. Here we established RAW264.7 macrophage cells stably expressing a luciferase reporter gene directed by iNOS or COX-2 promoter. Lipopolysaccharide (LPS) treatment stimulated the luciferase activity which paralleled with increased iNOS and COX-2 mRNA levels determined by RT-q-PCR. The LPS-stimulated luciferase activity was blocked by NF-£eB inhibitor CAPE and by nobiletin, an anti-inflammatory natural product from citrus peels. We have applied the platforms to screen various mushroom species; analysis by scatter plot revealed a strong correlation to the results obtained by ELISA-based detection of TNF-£\. Together we have established luciferase reporter systems sensitive to NF-£eB-dependent iNOS and COX-2 activation, which provides an alternative screening method for identifying food components with immune-modulatory activities. ? 2013 American Chemical Society. |
URI: | https://scholars.lib.ntu.edu.tw/handle/123456789/414032 | ISSN: | 00218561 | DOI: | 10.1021/jf404887u |
顯示於: | 食品科技研究所 |
在 IR 系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。