Figure 3.

TLR-4 mediates the mRNA expression levels of inflammatory cytokines in peritoneal macrophages and adhesion molecules in vascular endothelial cells from mice induced by anti-β2GPI–IgG. C3H/HeN mice (TLR-4 intact; n=8 in each group) and C3H/HeJ mice (TLR-4 defective; n=8 in each group) were treated by intraperitoneal injection of NR-IgG (100 µg antibody per injection), anti-β2GPI–IgG (100 µg antibody per injection) at time 0 and 48 h. The peritoneal macrophages and aortas were collected at 72 h after the first injection, and the positive control group was challenged with LPS (1 µg/g body weigh) 2 h before the experiments. Total RNA was extracted from peritoneal macrophages (2×10⁶) and mRNA expression levels of (A) TNF-α, (B) IL-1β and (C) IL-6 were detected by RT-qPCR. The total RNA was extracted from vascular tissue (aortas) and the mRNA levels of (D) VCAM-1, (E) ICAM-1 and (F) E-selectin were detected by RT-qPCR. The data are representative of three experiments. *P<0.05 vs. control NR-IgG; ^#P<0.05 vs. corresponding C3H/HeN stimulation group. TLR-4, Toll-like receptor-4; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; NR-IgG, isotype control antibody; Anti-β2GPI, anti-β2-glycoprotein I; LPS, lipopolysaccharide; TNF-α, tumor necrosis factor-α; IL, interleukin; ICAM-1, intercellular cell adhesion molecule-1; VCAM-1, vascular cell adhesion molecule-1.