Establishment and evaluation of biotechnological platform for screening health food with antiinflammation ability.
Chronic inflammation leads to a progressive inflammation in certain types of cells. Recent studies report that the activation of nuclear factor kappa B (NF-κB) increases the expression of inflammation-related protein such as inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2), which further enhance the chronic inflammation, thus conduct the development of disorders. The aim of the study is to develop an efficient method for screening food components with anti-inflammation function. Here we employed a reporter plasmid, which contains NF-κB response element followed by a minimal promoter for driving the down-stream luciferase reporter gene. After transfection of this plasmid to a mouse cell line RAW264.7, we obtained stable clones by using Hygromycin selection. Our results reveal that the luciferase activity of the cell based platform can be induced by the inflammation inducing reagent LPS and can be further suppressed by the administration of CAPE, an anti-inflammation chemical. The results estimated by our platform present good correlation to that analyzed by RT-Q-PCR. Additionally, the known anti-inflammation factors such as resveratrol, significantly counteracted the effect of LPS on our platform. Furthermore, the screening result of various mushroom extract showed that some fractions revealed NF-κB activating effects. Therefore, we conclude that the platform is effective in large scale screening for inflammatory regulating compounds.
Department of Biochemical Science and Technology, National Taiwan University, Taipei, Taiwan.
Wang-Ju
WJ
Hsieh
Institute of Food Science and Technology, National Taiwan University, Taipei, Taiwan.
Shih-Ting
ST
Chiou
Department of Seafood Science, National Kaoshuiung Marine University, Kaoshiung, Taiwan.
Min-Hsiung
MH
Pan
Institute of Food Science and Technology, National Taiwan University, Taipei, Taiwan.
Shu-Chen
SC
Hsieh
Cell platform; Inflammation; Luciferase assay; NF-κB