Overexpression of HMGB1 A-box reduced lipopolysaccharide-induced intestinal inflammation via HMGB1/TLR4 signaling in vitro

Figure 4 Expression of the HMGB1 protein and toll-like receptor 4 signaling pathways in lipopolysaccharide-stimulated SW480 cells. After pretreatment with EP (5 mmol/L, 1 h), the SW480 cells were treated with LPS (1 μg/mL, 24 h). The HMGB1 protein, TLR4, MYD88, and pNF-κB p65 mRNA and protein levels were determined by real-time PCR (A, C, E), densitometric quantification and representative western blotting with β-actin as the loading control (B, D, F, G, H). LPS increased the HMGB1, TLR4, MYD88, and pNF-κB p65 mRNA and protein levels in the LPS-stimulated SW480 cells compared with the LPS-untreated group, but EP downregulated these mRNA and protein levels in the LPS-stimulated SW480 cells. By contrast, HMGB1 A-box failed to downregulate the HMGB1, TLR4, MYD88, and pNF-κB p65 mRNA and protein levels compared with the mock-transfection group. The data are expressed as the mean ± SD and are derived from three independent experiments, which were each performed in duplicate. The means with letters above them are significantly different (^aP < 0.05, ^bP < 0.01 vs control).