title: Combination of antithrombin and recombinant thrombomodulin modulates neutrophil cell-death and decreases circulating DAMPs levels in endotoxemic rats.
abstract: BACKGROUND
  The activation of coagulation is recognized as a universal event in severe sepsis. Both antithrombin and thrombomodulin play pivotal roles as suppressors of coagulation. Since the levels of both anticoagulants decrease significantly, we hypothesized that a combination therapy would be beneficial.
  
  
  METHODS
  A sepsis model was established using the intravenous infusion of lipopolysaccharide (LPS). Either 125 IU/kg of antithrombin, 0.25mg/kg of recombinant thrombomodulin, or a combination of both agents was injected immediately after LPS infusion (n=7 each), while only a physiological saline was injected in the control group (n=7). Blood samples were obtained at eight hours after LPS infusion, and organ damage markers and the plasma levels of damage-associated molecular patterns (DAMPs), such as histone H3 and cell-free DNA (cf-DNA), were measured. In another series, the leukocytes harvested from normal rats were cultured in plasma obtained from each group (n=7). Eight hours later, the leukocytes were stained with green fluorescent protein, Annexin V and 7-AAD, and the proportion of alive+apoptic/necrotic cells was calculated.
  
  
  RESULTS
  Organ damage markers such as ALT and BUN were maintained best in the combination group (P<0.05). The circulating levels of histone H3 and cf-DNA were both significantly lower in the combination therapy group (P<0.01, 0.05, respectively). The proportion of alive+apoptic/necrotic cells was significantly higher in the combination therapy group (P<0.05).
  
  
  CONCLUSIONS
  The coadministration of antithrombin and recombinant thrombomodulin can modulate cell death and decrease the circulating levels of histone H3 and cf-DNA, leading to protection against organ damage in a rat model of sepsis.
authors.affiliation: Department of Emergency Disaster Medicine, Juntendo University Graduate School of Medicine. Electronic address: toshiiba@cf6.so-net.ne.jp.
authors.forename: Toshiaki
authors.initials: T
authors.lastname: Iba
authors.affiliation: Department of Emergency Disaster Medicine, Juntendo University Graduate School of Medicine. Electronic address: miki.takahiro@nihon-u.ac.jp.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Miki
authors.affiliation: Department of Emergency Disaster Medicine, Juntendo University Graduate School of Medicine. Electronic address: nhashigu@juntendo.ac.jp.
authors.forename: Naoyuki
authors.initials: N
authors.lastname: Hashiguchi
authors.affiliation: Department of Clinical Laboratory Medicine, Juntendo University Graduate School of Medicine. Electronic address: tabe@juntendo.ac.jp.
authors.forename: Yoko
authors.initials: Y
authors.lastname: Tabe
authors.affiliation: Department of Host Defense and Biochemical Research, Juntendo University Graduate School of Medicine. Electronic address: nagaokai@juntendo.ac.jp.
authors.forename: Isao
authors.initials: I
authors.lastname: Nagaoka
mesh_terms: D000818:Animals; D000991:Antithrombins / Q000494:pharmacology*; D001777:Blood Coagulation / Q000187:drug effects; D016923:Cell Death / Q000187:drug effects; D019446:Endotoxemia / Q000097:blood* / Q000188:drug therapy*; D018375:Neutrophil Activation / Q000187:drug effects; D009504:Neutrophils / Q000187:drug effects* / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D011994:Recombinant Proteins / Q000008:administration & dosage; D018805:Sepsis / Q000097:blood / Q000188:drug therapy; D018180:Thrombomodulin / Q000008:administration & dosage*
chemical_list: D000991:Antithrombins; D011994:Recombinant Proteins; D018180:Thrombomodulin
keywords: Antithrombin; Cell-free DNA; Damage-associated molecular patterns (DAMPs); Histone; Recombinant thrombomodulin; Sepsis