title: Real-time monitoring of inflammation status in 3T3-L1 adipocytes possessing a secretory Gaussia luciferase gene under the control of nuclear factor-kappa B response element.
abstract: We have established 3T3-L1 cells possessing a secretory Gaussia luciferase (GLuc) gene under the control of nuclear factor-kappa B (NF-κB) response element. The 3T3-L1 cells named 3T3-L1-NF-κB-RE-GLuc could differentiate into adipocyte as comparably as parental 3T3-L1 cells. Inflammatory cytokines such as tumor necrosis factor (TNF)-α and interleukin (IL)-1β induced GLuc secretion of 3T3-L1-NF-κB-RE-GLuc adipocytes in a concentration- and time-dependent manner. GLuc secretion of 3T3-L1-NF-κB-RE-GLuc adipocytes was also induced when cultured with RAW264.7 macrophages and was dramatically enhanced by lipopolysaccharide (LPS)-activated macrophages. An NF-κB activation inhibitor BAY-11-7085 and an antioxidant N-acetyl cysteine significantly suppressed GLuc secretion induced by macrophages. Finally, we found that rosemary-derived carnosic acid strongly suppressed GLuc secretion induced by macrophages and on the contrary up-regulated adiponectin secretion. Collectively, by using 3T3-L1-NF-κB-RE-GLuc adipocytes, inflammation status can be monitored in real time and inflammation-attenuating compounds can be screened more conveniently.
authors.affiliation: Department of Life Sciences, Graduate School of Bioresources, Mie University, Tsu 514-8507, Japan.
authors.forename: Haruka
authors.initials: H
authors.lastname: Nagasaki
authors.forename: Takeshi
authors.initials: T
authors.lastname: Yoshimura
authors.forename: Naohito
authors.initials: N
authors.lastname: Aoki
mesh_terms: D041721:3T3-L1 Cells; D017667:Adipocytes / Q000187:drug effects / Q000378:metabolism*; D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D016207:Cytokines / Q000235:genetics*; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D007249:Inflammation / Q000235:genetics*; D008156:Luciferases / Q000235:genetics*; D051379:Mice; D008991:Monitoring, Physiologic / Q000379:methods*; D016328:NF-kappa B / Q000378:metabolism; D020218:Response Elements; D013997:Time Factors
chemical_list: D000975:Antioxidants; D016207:Cytokines; D016328:NF-kappa B; D008156:Luciferases