Evaluation of the effect of recombinant thrombomodulin on a lipopolysaccharide-induced murine sepsis model

Abstract

Abstract.To evaluate the effect of recombinant human thrombomodulin (rTM) on sepsis, the levels of nucleosome as well as high-mobility group box 1 (HMGB1) and cytokines in sera and peritoneal fluids were measured in a mouse model of lipopolysaccharide (LPS)-induced sepsis after administration of rTM. C57BL/6 mice were intraperitoneally injected with LPS (15 mg/kg; Escherichia coli O111:B4) with or without the intravenous administration of rTM (3 mg/kg; 30 min prior to or 2 h after LPS injection). The survival rates were evaluated and levels of tumor necrosis factor (TNF)-α, interleukin (IL)-6, monocyte chemoattractant protein (MCP)-1, HMGB1 and nucleosome in sera and peritoneal fluids were analyzed by ELISA. Administration of rTM prior to or after LPS improved the survival rate of septic mice. In addition, rTM administered prior to or after LPS suppressed the level of pro-inflammatory cytokine TNF-α in sera at 1-3 h after LPS injection, whereas only the administration of rTM after LPS suppressed the levels of HMGB1 and nucleosome (late-phase mediators of sepsis) (9-12 h) in sera after the LPS injection. Furthermore, administration of rTM prior to or after LPS suppressed the level of TNF-α in the peritoneal fluids at 1-3 h after LPS injection, whereas only the administration of rTM after LPS suppressed the levels of IL-6 and MCP-1 in the peritoneal fluids at 6-9 h after LPS injection. These observations indicated that administration of rTM significantly improves the survival rate and suppresses the increased levels of TNF-α, IL-6, MCP-1, HMGB1 and nucleosome in the LPS-induced septic shock model. Thus, rTM may exert a protective action on sepsis and reduce mortality, possibly by reducing not only the levels of cytokines and chemokine but also the levels of late-phase mediators of sepsis.

Recombinant human thrombomodulin attenuated sepsis severity in a non-surgical preterm mouse model

“…Nagato et al reported that the intravenous administration of 1 mg/kg of rhTM 30 min prior to sepsis induction significantly improved sepsis survival, and suppressed inflammatory cytokines and HMGB1 elevation in an LPS-treated rat sepsis model 16 . Takehara et al reported that the intravenous administration of 3 mg/kg of rhTM 30 min prior to sepsis induction improved sepsis survival, and suppressed the elevation of inflammatory cytokines and HMGB1 in serum and ascites in an LPS-treated mouse sepsis model 25 . However, these studies used adult rodent sepsis models, and there has been no report investigating the effects of rhTM in an animal model of neonatal sepsis.…”

Thrombomodulin as a Physiological Modulator of Intravascular Injury

“…In a mouse histone-induced septic model, pretreatment with rTM reduced mortality rates by neutralizing histones ( 20 ). In a rat sepsis/peritonitis model ( 33 ) and a murine LPS-induced septic model ( 34 ), rTM controlled sepsis-related immunothrombosis by limiting abnormal hemostasis and NET formation.…”

Recombinant Human Soluble Thrombomodulin Suppresses Monocyte Adhesion by Reducing Lipopolysaccharide-Induced Endothelial Cellular Stiffening

“…Although the anti-inflammatory and cytoprotective effects deployed by rsTM to counter inflamed endothelial cells have been known [3,5,6], the impact of rsTM on endothelial cellular stiffening under inflammatory conditions remains elusive. Therefore, we investigated whether rsTM suppresses endothelial cellular stiffening during LPS-mediated sterile inflammation.…”

“…Thus, endothelial cell dysfunction is a cause of the vascular lesions associated with functional and structural abnormalities in vascular vessels. Thrombomodulin (TM) is an anticoagulant protein that is predominantly expressed on the surface of endothelial cells [1][2][3]. TM directly binds to thrombin and inhibits thrombin procoagulant activity [1,2].…”