RRC ID 18610
Author Sakamoto T, Yamaguchi Y, Goto T, Takahashi N, Kawada T.
Title An in vitro analysis system using a fluorescence protein reporter for evaluating anti-inflammatory effects in macrophages.
Journal Biosci. Biotechnol. Biochem.
Abstract 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.
Volume 75(8)
Pages 1582-7
Published 2011
DOI 10.1271/bbb.110278
PII JST.JSTAGE/bbb/110278
PMID 21821941
MeSH 3T3-L1 Cells Adipocytes / cytology Adipocytes / drug effects Adipocytes / metabolism* Animals Anti-Inflammatory Agents / analysis* Anti-Inflammatory Agents / pharmacology Anti-Inflammatory Agents / therapeutic use Biological Assay* Chemokine CCL2 / chemistry Chemokine CCL2 / genetics* Coculture Techniques Culture Media, Conditioned / pharmacology Fluorescence Gene Expression Genes, Reporter Green Fluorescent Proteins / analysis* Green Fluorescent Proteins / genetics Green Fluorescent Proteins / immunology Humans Inflammation / drug therapy Inflammation / immunology Inflammation / metabolism Inflammation / pathology Insulin Resistance / immunology Lipopolysaccharides / pharmacology Macrophages / cytology Macrophages / drug effects Macrophages / metabolism* Mice Obesity / drug therapy Obesity / immunology Obesity / metabolism Obesity / pathology Promoter Regions, Genetic RNA, Messenger / analysis RNA, Messenger / biosynthesis Recombinant Fusion Proteins / analysis* Recombinant Fusion Proteins / genetics Recombinant Fusion Proteins / immunology Tumor Necrosis Factor-alpha / chemistry Tumor Necrosis Factor-alpha / genetics*
IF 1.255
Times Cited 2
Human and Animal Cells RAW 264 (RCB0535)