An in Vitro Analysis System Using a Fluorescence Protein Reporter for Evaluating Anti-Inflammatory Effects in Macrophages
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概要
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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 J2, 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.
著者
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TAKAHASHI Nobuyuki
Laboratory for Molecular Function of Food, Division of Food Science and Biotechnology, Graduate Scho
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Yamada Yukio
Mechanical Engineering Laboratory Material Science And Bioengineering Department Aist
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YAMAGUCHI Yuko
Laboratory of Physics,Faculty of Education,Chiba University
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Kawada Teruo
Laboratory For Molecular Function Of Food Division Of Food Science And Biotechnology Graduate School
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Goto Tsuyoshi
Laboratory For Molecular Function Of Food Division Of Food Science And Biotechnology Graduate School
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Yamada Yuko
Kyoto Univ. Kyoto Jpn
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SAKAMOTO Tomoya
Laboratory for Molecular Functions of Food, Division of Food Science and Biotechnology, Graduate Sch
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