Abstract
Anti‑phospholipid syndrome (APS) is a systematic autoimmune disease that is associated with presence of antiphospholipid antibodies (aPL), recurrent thrombosis, and fetal morbidity in pregnancy. Toll‑like receptor‑4 (TLR‑4), a member of TLR family, is known to have a fundamental role in pathogen recognition and activation of innate immunity. The β2‑glycoprotein I (β2GPI), a protein circulating in the blood at a high concentration, is able of scavenging lipopolysaccharide (LPS) and clear unwanted anionic cellular remnants, such as microparticles, from the circulation. Our previous study demonstrated that TLR‑4 and its signaling pathways contribute to the upregulation of pro‑coagulant factors and pro‑inflammatory cytokines in monocytes induced by anti‑β2GPI in vitro. The present study aimed to define the roles of TLR‑4 in vivo. C3H/HeN mice (TLR‑4 intact) and C3H/HeJ mice (TLR‑4 defective) were stimulated with an intraperitoneal injection with anti‑β2GPI‑immunoglobulin G(IgG), then peritoneal macrophages and vascular endothelial cells (VECs) were extracted from treated mice, and analyses were conducted on the expression profiles of pro‑inflammatory cytokines and adhesion molecules. The results demonstrated that the expression of pro‑inflammatory cytokines, including tumor necrosis factor‑α (TNF‑α), interleukin (IL)‑1β and IL‑6, in the peritoneal macrophages, and adhesion molecules, including intercellular cell adhesion molecule‑1 (ICAM‑1), vascular cell adhesion molecule‑1 (VCAM‑1) and E‑selectin, in VECs of C3H/HeN mice (TLR‑4 intact) were significantly higher than those of C3H/HeJ mice (TLR‑4 defective). The phosphorylation levels of p38 mitogen‑activated protein kinase (MAPK) and nuclear factor‑κB (NF‑κB) p65 in peritoneal macrophages and VECs from C3H/HeN mice stimulated with anti‑β2GPI‑IgG were significantly increased compared with those from C3H/HeJ mice (TLR‑4 defective). The isotype control antibody (NR‑IgG) had no such effects on peritoneal macrophages and VECs. Furthermore, the inhibitors of TLR‑4, p38 MAPK and NF‑κB may significantly reduce the anti‑β2GPI‑IgG‑induced TNF‑α, IL‑1β and IL‑6 mRNAs expression in the peritoneal macrophages from TLR‑4 intact mice. The results indicated that a TLR‑4 signal transduction pathway is involved in anti‑β2GPI‑IgG‑induced activation of peritoneal macrophages and VECs. This study has provided a basis for subsequent investigations to elucidate the pathological mechanisms underlying anti‑phospholipid syndrome.