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. 2012 Jul 2;7(7):e40056. doi: 10.1371/journal.pone.0040056

Figure 5. Effect of 11β-HSD1 on the inflammation gene expression in PA or LPS-activated J774A.1 macrophages in vitro.

Figure 5

A, J774.1 macrophages were treated with palmitic acid (PA) (50−200 µmol/L) or LPS (100 ng/ml) for 24 h. B–C, J774.1 macrophages were activated by PA (200 µmol/L) or LPS (100 ng/ml) and co-treated with 11β-HSD1 inhibitor BVT.2733 (25−100 µmol/L) for 24 h. D–G, J774.1 macrophages were transfected with either sh-RNA for mouse 11β-HSD1 (sh-HSD1) or a negative control (sh-con) by Lentivirus. After 72 h incubation, cells were treated with PA (200 µmol/L) or LPS (100 ng/ml) for 24 h. Efficiency of 11β-HSD1 knockdown on mRNA level (D) and protein level (E). Effects of knockdown of 11β-HSD1 on MCP-1, IL-6, and TNF-α expression in PA (F) or LPS (G) treated macrophages. H–K, J774.1 macrophages were transfected with the expression vector for 11β-HSD1 (HSD1) or a corresponding empty vector (emp) using Lentivirus. After 72 h incubation, cells were treated with PA (100 µmol/L) or LPS (50 ng/ml) and co-treated with 11β-HSD1 inhibitor BVT.2733 for 24 h. Efficiency of 11β-HSD1 overexpression on mRNA level (H) and protein level (I). Effects of overexpression of 11β-HSD1 on MCP-1, IL-6, and TNF-α expression in PA (J) or LPS (K) treated macrophages. mRNA for IL-6, MCP-1 and TNF-α were determined by real-time PCR, protein of 11β-HSD1 were determined by Western blot. The results are shown as the means ± SEM of three individual experiments. *P<0.05; **P<0.01 vs con (B, C, F, G) or sh-con (D) or emp (H) or emp+PA (J) or emp+LPS (K). # P<0.05; ## P<0.01 vs PA (B) or LPS (C) or sh-con+PA (F) or sh-con+LPS (G) or HSD1+PA (J) or HSD1+LPS (K).