PMID- 21821941
OWN - NLM
STAT- MEDLINE
DCOM- 20120117
LR  - 20110826
IS  - 1347-6947 (Electronic)
IS  - 0916-8451 (Linking)
VI  - 75
IP  - 8
DP  - 2011
TI  - An in vitro analysis system using a fluorescence protein reporter for evaluating 
      anti-inflammatory effects in macrophages.
PG  - 1582-7
AB  - 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-alpha (TNFalpha). 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-12,14Delta-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.
FAU - Sakamoto, Tomoya
AU  - Sakamoto T
AD  - Laboratory for Molecular Functions of Food, Division of Food Science and 
      Biotechnology, Graduate School of Agriculture, Kyoto University, Uji, Kyoto, 
      Japan.
FAU - Yamaguchi, Yuko
AU  - Yamaguchi Y
FAU - Goto, Tsuyoshi
AU  - Goto T
FAU - Takahashi, Nobuyuki
AU  - Takahashi N
FAU - Kawada, Teruo
AU  - Kawada T
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20110807
PL  - England
TA  - Biosci Biotechnol Biochem
JT  - Bioscience, biotechnology, and biochemistry
JID - 9205717
RN  - 0 (Anti-Inflammatory Agents)
RN  - 0 (Chemokine CCL2)
RN  - 0 (Culture Media, Conditioned)
RN  - 0 (Lipopolysaccharides)
RN  - 0 (RNA, Messenger)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 0 (Tumor Necrosis Factor-alpha)
RN  - 147336-22-9 (Green Fluorescent Proteins)
SB  - IM
MH  - 3T3-L1 Cells
MH  - Adipocytes/cytology/drug effects/*metabolism
MH  - Animals
MH  - Anti-Inflammatory Agents/*analysis/pharmacology/therapeutic use
MH  - *Biological Assay
MH  - Chemokine CCL2/chemistry/*genetics
MH  - Coculture Techniques
MH  - Culture Media, Conditioned/pharmacology
MH  - Fluorescence
MH  - Gene Expression
MH  - Genes, Reporter
MH  - Green Fluorescent Proteins/*analysis/genetics/immunology
MH  - Humans
MH  - Inflammation/drug therapy/immunology/metabolism/pathology
MH  - Insulin Resistance/immunology
MH  - Lipopolysaccharides/pharmacology
MH  - Macrophages/cytology/drug effects/*metabolism
MH  - Mice
MH  - Obesity/drug therapy/immunology/metabolism/pathology
MH  - Promoter Regions, Genetic
MH  - RNA, Messenger/analysis/biosynthesis
MH  - Recombinant Fusion Proteins/*analysis/genetics/immunology
MH  - Tumor Necrosis Factor-alpha/chemistry/*genetics
EDAT- 2011/08/09 06:00
MHDA- 2012/01/18 06:00
CRDT- 2011/08/09 06:00
PHST- 2011/08/09 06:00 [entrez]
PHST- 2011/08/09 06:00 [pubmed]
PHST- 2012/01/18 06:00 [medline]
AID - JST.JSTAGE/bbb/110278 [pii]
AID - 10.1271/bbb.110278 [doi]
PST - ppublish
SO  - Biosci Biotechnol Biochem. 2011;75(8):1582-7. doi: 10.1271/bbb.110278. Epub 2011 
      Aug 7.