Combination of antithrombin and recombinant thrombomodulin attenuates leukocyte-endothelial interaction and suppresses the increase of intrinsic damage-associated molecular patterns in endotoxemic rats.

INTRODUCTION

Both antithrombin (AT) and thrombomodulin are key players in physiological anticoagulant systems. Because the levels of both factors are known to decrease significantly during severe sepsis, we hypothesized that a combination therapy would be effective.

METHODS

A sepsis model was established using the intravenous infusion of lipopolysaccharide (LPS). A dose of 125 IU/kg of AT, 0.25 mg/kg of recombinant thrombomodulin, or a combination of both agents was injected immediately after LPS infusion (n = 7, each). Intravital observation of the mesenteric microcirculation was performed, and leukocyte adhesion and blood flow were calculated at 3 h after LPS infusion. Immediately after the observation, blood samples were obtained and coagulation markers, organ damage markers, the circulating levels of nucleosome and high-mobility group box 1 were measured.

RESULTS

Microscopic findings revealed the suppression of leukocyte adhesion and thrombus formation in the combination group. The number of adhesive leukocytes on the endothelium was significantly suppressed (P < 0.01), and the blood flow in venules was better maintained in the combination group compared with the placebo control (P < 0.01). The blood samples showed the suppressed activation in coagulation, no significant changes were observed in the organ damage markers in the treatment groups. The circulating levels of nucleosome and high-mobility group box 1 were both decreased significantly in the combination group compared with the placebo control (P < 0.01).

CONCLUSIONS

The coadministration of AT and recombinant thrombomodulin is effective for the suppression of leukocyte activation and cell death during sepsis.