Establishment of Reporter Platforms Capable of Detecting NF-κB Mediated Immuno-Modulatory Activity
| Autor: | Ting-Jang Lu, Chung-Huang Wang, Shuchen Hsieh, Shih-Ting Chiou, Ju-Hwa Lin, Min-Hsiung Pan, Yi-An Chen, An-Na Chiang |
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| Rok vydání: | 2013 |
| Předmět: |
Citrus
biology Plant Extracts Chemistry Macrophages Anti-Inflammatory Agents NF-kappa B Nitric Oxide Synthase Type II NF-κB General Chemistry Molecular biology Nuclear factor kappa b Nitric oxide synthase Mice chemistry.chemical_compound Cyclooxygenase 2 Genes Reporter biology.protein Animals Biological Assay Luciferase General Agricultural and Biological Sciences |
| Zdroj: | Journal of Agricultural and Food Chemistry. 61:12582-12587 |
| ISSN: | 1520-5118 0021-8561 |
| DOI: | 10.1021/jf404887u |
| Popis: | Cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) are transcriptional targets of nuclear factor kappa B (NF-κB) 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-κB 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-α. Together we have established luciferase reporter systems sensitive to NF-κB-dependent iNOS and COX-2 activation, which provides an alternative screening method for identifying food components with immune-modulatory activities. |
| Databáze: | OpenAIRE |
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