DNA microarray analyses of genes expressed differentially in 3T3-L1 adipocytes co-cultured with murine macrophage cell line RAW264.7 in the presence of the toll-like receptor 4 ligand bacterial endotoxin.
Recent studies have suggested that macrophages were integrated into adipose tissues to interact with adipocytes, thereby exacerbating inflammatory responses. Furthermore, both adipocytes and macrophages appear to express toll-like receptor-4 (TLR-4), and free fatty acids may stimulate cells through TLR-4. Herein, we analyzed genes differentially expressed in adipocytes when co-cultured with macrophages in the presence of a ligand for TLR-4, bacterial lipopolysaccharide (LPS). RAW264.7, a murine macrophage cell line and differentiated 3T3-L1 adipocytes were co-cultured using a transwell system. Genes differentially expressed in adipocytes were analyzed by the DNA microarray method following 4, 8, 12 and 24 h stimulation with 1 ng ml(-1) of Escherichia coli LPS. Randomly selected genes with high expressions were confirmed by quantitative methods at both the gene and the protein level. Co-culture of macrophages and adipocytes with a low LPS concentration (1 ng ml(-1)) markedly upregulated gene expressions associated with inflammation and/or angiogenesis, such as those of interleukin-6 (IL-6), MCP-1, RANTES and CXCL1/KC, in adipocytes. Furthermore, several genes associated with insulin resistance were differentially expressed. Upregulations of genes encoding MCP-1, RANTES and CXC/KC were confirmed by quantitative methods. These results suggest that ligands for TLR-4 stimulate both adipocytes and macrophages to upregulate the expressions of many genes associated with inflammation and/or angiogenesis.
Yamashita
A
A
Department of Dental Science for Health Promotion, Hiroshima University Graduate School of Biomedical Sciences, Hiroshima, Japan.
Soga
Y
Y
Iwamoto
Y
Y
Asano
T
T
Li
Y
Y
Abiko
Y
Y
Nishimura
F
F
D041721:3T3-L1 Cells / Q000378:metabolism; D017667:Adipocytes / Q000276:immunology*; D000818:Animals; D002460:Cell Line / Q000378:metabolism; D004731:Endotoxins / Q000235:genetics / Q000276:immunology*; D015870:Gene Expression; D008264:Macrophages / Q000276:immunology*; D051379:Mice; D046228:Microarray Analysis; D051197:Toll-Like Receptor 4 / Q000235:genetics / Q000276:immunology*; D015854:Up-Regulation
D004731:Endotoxins; C495345:TLR4 protein, human; D051197:Toll-Like Receptor 4