Figure 8

Effect of 67-kDa laminin receptor neutralization on (−)-epigallocatechin gallate-mediated inhibition of lipopolysaccharide actions. (A,B) Effect of 67-kDa laminin receptor (67LR) antibody on (−)-epigallocatechin gallate (EGCG)-mediated suppression of the induction of TNF-α and IL-1β by lipopolysaccharide (LPS). Human cerebral microvascular endothelial cells/D3 (hCMEC/D3) were treated with antibody against 67LR (5 μg/ml) or control IgM for 1 h prior to LPS and/or EGCG treatment. RNA extracted was subjected to real time PCR for TNF-α and IL-1β expression. (C) Effect of 67LR antibody on EGCG-mediated inhibition of the induction of NF-κB activity by LPS. Cells were transfected with pNF-κB-Luc plasmid and then treated with antibody against 67LR (5 μg/ml) or control IgM for 1 h prior to LPS and/or EGCG treatment for an additional 6 h. Luciferase activity was measured in the cell lysates. (D) Effect of 67LR antibody on EGCG-mediated suppression of transmembrane diffusion by LPS. The cells were treated with 67LR antibody, EGCG and LPS as described above. The apical apartment was then loaded with fluorescin sodium salt and the fluorescence in the basal apartment was measured within 1 h. Data are expressed as mean ± SD of three independent experiments. (*P < 0.05, **P < 0.01, compared with LPS treatment only).