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
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
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DOI：
10.1038/ijo.2008.153
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发表时间：
2008-11-01
影响因子：
4.9
通讯作者：
Nishimura, F.
中科院分区：
文献类型：
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作者：
Yamashita, A.;Soga, Y.;Nishimura, F.
最近的研究表明，巨噬细胞整合到脂肪组织中，与脂肪细胞相互作用，从而加剧炎症反应。此外，脂肪细胞和巨噬细胞似乎都表达 Toll 样受体 4 (TLR-4)，游离脂肪酸可能通过 TLR-4 刺激细胞。在此，我们分析了在 TLR-4 配体（细菌脂多糖 (LPS)）存在下与巨噬细胞共培养时脂肪细胞中差异表达的基因。使用 Transwell 系统共培养小鼠巨噬细胞系 RAW264.7 和分化的 3T3-L1 脂肪细胞。用1 ng ml(-1)大肠杆菌LPS刺激4、8、12和24 h后，通过DNA微阵列法分析脂肪细胞中差异表达的基因。随机选择的高表达基因通过基因和蛋白质水平的定量方法进行确认。巨噬细胞和脂肪细胞与低 LPS 浓度 (1 ng ml(-1)) 的共培养显着上调脂肪细胞中与炎症和/或血管生成相关的基因表达，例如白细胞介素 6 (IL-6)、MCP-1、RANTES 和 CXCL1/KC。此外，与胰岛素抵抗相关的几个基因也存在差异表达。通过定量方法证实了编码 MCP-1、RANTES 和 CXC/KC 的基因的上调。这些结果表明，TLR-4 的配体刺激脂肪细胞和巨噬细胞上调许多与炎症和/或血管生成相关的基因的表达。
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.