Inflammation Research

, Volume 59, Issue 7, pp 511–518 | Cite as

High-dose antithrombin III prevents heat stroke by attenuating systemic inflammation in rats

  • Satoshi Hagiwara
  • Hideo Iwasaka
  • Chihiro Shingu
  • Shigekiyo Matsumoto
  • Tomohisa Uchida
  • Takayuki Noguchi
Original Research Paper



Systemic inflammatory mediators, including the high mobility group box 1 (HMGB1) protein, play important roles in the development of various inflammatory conditions. Although anticoagulants, such as antithrombin III (AT III), inhibit inflammation resulting from various causes, their anti-inflammatory mechanism of action is not well understood. Nevertheless, as heat stroke is a severe inflammatory response disease, we hypothesized that AT III would inhibit inflammation and prevent heat stress-induced acute heat stroke.


Male Wistar rats received a bolus injection of saline or 250 U of AT III per kg of body weight into the tail vein, followed by heat stress (exposure to 42°C for 30 min). Levels of cytokines (interleukin-1 β, interleukin-6, and TNF-α), NOx, and HMGB1 were measured in serum and tissue at regular intervals for 6 h after the heat stress induction.


Levels of cytokines, NOx, and HMGB1 in serum decreased over time in AT III-treated rats. AT III pretreatment also reduced NOx levels during heat stress-induced inflammation. As a result, AT III pretreatment improved survival in a rat model of heat stress-induced acute inflammation.


Our data suggest that AT III pretreatment inhibited the secretion of cytokines, NOx, and HMGB1, and prevented heat stress-induced acute inflammation.


Inflammation Cytokine Nitric oxide Anticoagulant Heat stress 



The authors wish to thank Hiroaki Kawazato and Aiko Yasuda for helpful advice on hematoxylin and eosin staining.


  1. 1.
    Grogan H, Hopkins PM. Heat stroke: implications for critical care and anaesthesia. Br J Anaesth. 2002;88(5):700–7.CrossRefPubMedGoogle Scholar
  2. 2.
    Bouchama A, Knochel JP. Heat stroke. N Engl J Med. 2002;346:1978–88.CrossRefPubMedGoogle Scholar
  3. 3.
    Simon HB. Hyperthermia. N Engl J Med. 1993;329:483–7.CrossRefPubMedGoogle Scholar
  4. 4.
    Lee JJ, Lin MT, Wang NL, Lin CL, Chang CK. Platonin, a cyanine photosensitizing dye, causes attenuation of circulatory shock, hypercoagulable state, and tissue ischemia during heat stroke. Shock. 2005;24:577–82.CrossRefPubMedGoogle Scholar
  5. 5.
    Lu KC, Wang JY, Lin SH, Chu P, Lin YF. Role of circulating cytokines and chemokines in exertional heatstroke. Crit Care Med. 2004;32:399–403.CrossRefPubMedGoogle Scholar
  6. 6.
    Bouchama A, Knochel JP. Heat stroke. N Engl J Med. 2002;346:1978–88.CrossRefPubMedGoogle Scholar
  7. 7.
    Moncada S, Randomashi NW, Palmer RMJ. Endothelium-derived relaxing factor: identification as nitric oxide and role in control of vascular tone and platelet function. Biochem Pharmacol. 1988;37:2495–501.CrossRefPubMedGoogle Scholar
  8. 8.
    Alzeer AH, Al-Arifi A, Warsy AS, Ansari Z, Zhang H, Vincent JL. Nitric oxide production is enhanced in patients with heat stroke. Intensive Care Med. 1999;25:58–62.CrossRefPubMedGoogle Scholar
  9. 9.
    Bustin M. Regulation of DNA-dependent activities by the functional motifs of the high-mobility-group chromosomal proteins. Mol Cell Biol. 1999;19:5237–46.PubMedGoogle Scholar
  10. 10.
    Wang H, Bloom O, Zhang M, Vishnubhakat JM, Ombrellino M, Che J, et al. HMG-1 as a late mediator of endotoxin lethality in mice. Science. 1999;285:248–51.CrossRefPubMedGoogle Scholar
  11. 11.
    Levy RM, Mollen KP, Prince JM, Kaczorowski DJ, Vallabhaneni R, Liu S, et al. Systemic inflammation and remote organ injury following trauma require HMGB1. Am J Physiol Regul Integr Comp Physiol. 2007;293:R1538–44.PubMedGoogle Scholar
  12. 12.
    Tsung A, Hoffman RA, Izuishi K, Critchlow ND, Nakao A, Chan MH, et al. Hepatic ischemia/reperfusion injury involves functional TLR4 signaling in nonparenchymal cells. J Immunol. 2005;175:7661–8.PubMedGoogle Scholar
  13. 13.
    Opal SM, Kessler CM, Roemisch J, Knaub S. Antithrombin, heparin, and heparan sulfate. Crit Care Med. 2002;30:S325–31.CrossRefPubMedGoogle Scholar
  14. 14.
    Taylor FB Jr, Emerson TE Jr, Jordan R, Chang AK, Blick KE. Antithrombin-III prevents the lethal effects of Escherichia coli infusion in baboons. Circ Shock. 1988;26:227–35.PubMedGoogle Scholar
  15. 15.
    Wiedermann CJ. Clinical review: molecular mechanisms underlying the role of antithrombin in sepsis. Crit Care. 2006;10:209.CrossRefPubMedGoogle Scholar
  16. 16.
    Hagiwara S, Iwasaka H, Matsumoto S, Noguchi T. High dose antithrombin III inhibits HMGB1 and improves endotoxin-induced acute lung injury in rats. Intensive Care Med. 2008;34:361–7.CrossRefPubMedGoogle Scholar
  17. 17.
    Murakami K, McGuire R, Cox RA, Jodoin JM, Bjertnaes LJ, Katahira J, et al. Heparin nebulization attenuates acute lung injury in sepsis following smoke inhalation in sheep. Shock. 2002;18:236–41.CrossRefPubMedGoogle Scholar
  18. 18.
    Heijnen BH, Straatsburg IH, Gouma DJ, Van Gulik TM. Decrease in core liver temperature with 10°C by in situ hypothermic perfusion under total hepatic vascular exclusion reduces liver ischemia and reperfusion injury during partial hepatectomy in pigs. Surgery. 2003;134:806–17.CrossRefPubMedGoogle Scholar
  19. 19.
    Chiu CJ, McArdle AH, Brown R, Scott HJ, Gurd FN. Intestinal mucosal lesion in low-flow states: I, a morphological, hemodynamic, and metabolic reappraisal. Arch Surg. 1970;101:478–83.PubMedGoogle Scholar
  20. 20.
    Green LC, Wagner DA, Glogowski J, Skipper PL, Wishnok JS, Tannenbaum SR. Analysis of nitrate, nitrite and [15 N] nitrate in biological fluids. Anal Biochem. 1982;126:131–8.CrossRefPubMedGoogle Scholar
  21. 21.
    Voll RE, Urbonaviciute V, Herrmann M, Kalden JR. High mobility group box 1 in the pathogenesis of inflammatory and autoimmune diseases. Isr Med Assoc J. 2008;10:26–8.PubMedGoogle Scholar
  22. 22.
    Mantell LL, Parrish WR, Ulloa L. Hmgb-1 as a therapeutic target for infectious and inflammatory disorders. Shock. 2006;25:4–11.CrossRefPubMedGoogle Scholar
  23. 23.
    Lin XJ, Li YL, Mei GP, Zou F, He DD, Liu XQ, Li YJ, Zhao TB, Lin MT. Activated protein c can be used as a prophylactic as well as a therapeutic agent for heat stroke in rodents. Shock. 2009 Mar 13.Google Scholar
  24. 24.
    Wiedermann CJ, Hoffmann JN, Juers M, Ostermann H, Kienast J, Briegel J, et al. High-dose antithrombin III in the treatment of severe sepsis in patients with a high risk of death: efficacy and safety. Crit Care Med. 2006;34:285–92.CrossRefPubMedGoogle Scholar
  25. 25.
    Dabrowski A, Gabryelewicz A. Nitric oxide contributes to multiorgan oxidative stress in acute pancreatitis. Scand J Gastroenterol. 1994;29:943–8.CrossRefPubMedGoogle Scholar
  26. 26.
    Viola G, al-Mufti RA, Sohail M, Williamson RC, Mathie RT. Nitric oxide induction in a rat model of selective pancreatic ischemia and reperfusion. Hepatogastroenterology. 2000;47:1250–5.PubMedGoogle Scholar
  27. 27.
    Jiang W, Pisetsky DS. The role of IFN-alpha and nitric oxide in the release of HMGB1 by RAW 264.7 cells stimulated with polyinosinic-polycytidylic acid or lipopolysaccharide. J Immunol. 2006;177:3337–43.PubMedGoogle Scholar
  28. 28.
    Hall DM, Buettner GR, Matthes RD, Gisolfi CV. Hyperthermia stimulates nitric oxide formation: electron paramagnetic resonance detection of NO-heme in blood. J Appl Physiol. 1994;77:548–53.PubMedGoogle Scholar
  29. 29.
    al-Mashhadani SA, Gader AG, al Harthi SS, Kangav D, Shaheen FA, Bogus F. The coagulopathy of heatstroke: alterations in coagulation and fibrinolysis in heatstroke patients during the pilgrimage (Haj) to Makkah. Blood Coagul Fibrinolysis. 1994;5:731–6.CrossRefPubMedGoogle Scholar
  30. 30.
    Bouchama A, Bridey F, Hammami MM, Lacombe C, al-Shail E, al-Ohali Y, et al. Activation of coagulation and fibrinolysis in heatstroke. Thromb Haemost. 1996;76:909–15.PubMedGoogle Scholar

Copyright information

© Birkhäuser Verlag, Basel/Switzerland 2010

Authors and Affiliations

  • Satoshi Hagiwara
    • 1
  • Hideo Iwasaka
    • 1
  • Chihiro Shingu
    • 1
  • Shigekiyo Matsumoto
    • 1
  • Tomohisa Uchida
    • 2
  • Takayuki Noguchi
    • 1
  1. 1.Department of Anesthesiology and Intensive Care MedicineOita University Faculty of MedicineYufu CityJapan
  2. 2.Department of Molecular PathologyOita University Faculty of MedicineYufu CityJapan

Personalised recommendations