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An in Vitro Analysis System Using a Fluorescence Protein Reporter for Evaluating Anti-Inflammatory Effects in Macrophages
Ist Teil von
Bioscience, biotechnology, and biochemistry, 2011-08, Vol.75 (8), p.1582-1587
Ort / Verlag
Tokyo: Japan Society for Bioscience, Biotechnology, and Agrochemistry
Erscheinungsjahr
2011
Link zum Volltext
Quelle
EZB Free E-Journals
Beschreibungen/Notizen
Monitoring of inflammation in adipose tissues, which causes insulin resistance, is valuable in evaluating insulin resistance. We developed an in vitro analysis system using a fluorescence protein (FP) as a reporter gene driven by pro-inflammatory cytokine promoters such as monocyte chemoattractant protein-1 (MCP-1) and tumor necrosis factor-α (TNFα). In the reporter-transfected RAW264 cells, the protein expression levels of green fluorescence protein (GFP) were increased by inflammatory stimulations such as lipopolysaccharide (LPS), conditioned medium prepared using hypertrophied 3T3-L1 adipocytes, and a co-culture system. The changes in fluorescence intensity were equivalent to those of the mRNA and protein expression levels for each cytokine. Moreover, the effects of 15-deoxy-12,14Δ-prostaglandine J
2
, a natural anti-inflammatory compound, were detectable in this system. These data indicate that the FP system developed here is an analysis system of low cost with simple procedures for evaluating inflammation, suggesting usability in the large-scale screening of anti-inflammatory compounds.