An in vitro analysis system using a fluorescence protein reporter for evaluating anti-inflammatory effects in macrophages.
Biosci Biotechnol Biochem
; 75(8): 1582-7, 2011.
Article
em En
| MEDLINE
| ID: mdl-21821941
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.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Bioensaio
/
Proteínas Recombinantes de Fusão
/
Fator de Necrose Tumoral alfa
/
Adipócitos
/
Quimiocina CCL2
/
Proteínas de Fluorescência Verde
/
Macrófagos
/
Anti-Inflamatórios
Idioma:
En
Revista:
Biosci Biotechnol Biochem
Assunto da revista:
BIOQUIMICA
/
BIOTECNOLOGIA
Ano de publicação:
2011
Tipo de documento:
Article
País de afiliação:
Japão