High dose antithrombin III inhibits HMGB1 and improves endotoxin-induced acute lung injury in rats.
OBJECTIVE
High mobility group box 1 (HMGB1) is an important factor in the development of sepsis. Previous work suggests that antithrombin III (ATIII) inhibits inflammation, but the mechanism of action is still poorly understood.


METHODS
Prospective controlled animal study in a university laboratory.


METHODS
Rats were randomly divided into a lipopolysaccharide (LPS)-induced sepsis control group and an ATIII-treated experimental group. Animals in the experimental group received a bolus of 250 units/kg of ATIII injected into the tail vein.


RESULTS
Animals receiving high-dose ATIII (250 units/kg) had significantly improved lung histopathology and survival compared to the control rats. We measured serum and lung levels of various cytokines and HMGB1 at regular intervals from 0 to 12 h after the induction of sepsis and demonstrated lower HMGB1 levels over time in ATIII-treated animals. In an in vitro experiment, we stimulated the mouse macrophage cell line RAW 264.7 with LPS in the presence or absence of ATIII. Subsequent measurement of HMGB1 concentrations in the supernatant and cell signaling molecules in cell lysates revealed an ATIII dose-dependent decrease in HMGB1 release. Furthermore, inhibition of IkB and p42 phosphorylation was observed with the administration of ATIII, suggestive of downstream signaling pathways.


CONCLUSIONS
High-dose ATIII decreases lung pathology and reduces mortality in a rat sepsis model. This finding may be mediated by the inhibition of HMGB1.
Hagiwara
Satoshi
S
Oita University Faculty of Medicine, Department of Brain and Nerve Science, Anesthesiology, 1-1 Idaigaoka, Hasamamachi, Yufu City, Oita 879-5593, Japan. saku@med.oita-u.ac.jp
Iwasaka
Hideo
H
Matsumoto
Shigekiyo
S
Noguchi
Takayuki
T
D000704:Analysis of Variance; D000818:Animals; D000990:Antithrombin III / Q000494:pharmacology*; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D024243:HMGB1 Protein / Q000187:drug effects*; D007124:Immunoenzyme Techniques; D008070:Lipopolysaccharides; D008297:Male; D011446:Prospective Studies; D011897:Random Allocation; D051381:Rats; D017208:Rats, Wistar; D012128:Respiratory Distress Syndrome / Q000188:drug therapy*
D024243:HMGB1 Protein; D008070:Lipopolysaccharides; D000990:Antithrombin III