Recombinant human soluble thrombomodulin reduces the severity and incidence of necrotizing enterocolitis in a newborn rat model

  • Bo Li
  • Ryuta Saka
  • Yuichi Takama
  • Takehisa Ueno
  • Yuko Tazuke
  • Hiroomi OkuyamaEmail author
Original Article



Necrotizing enterocolitis (NEC) remains the leading cause of death in preterm infants. Recombinant human soluble thrombomodulin (rTM) has been reported to have anti-inflammatory effects as well as antithrombogenic effects. The aim of this study was to evaluate the effect of rTM in a rat NEC model.


NEC was induced by enteral feeding with hyperosmolar formula, gavage administration of lipopolysaccharide and asphyxia stress. Controls were fed by their mother ad libitum. In the treatment group, rTM was administered subcutaneously twice (once each on the first and second day). All animals surviving beyond 96 h or that developed signs of distress were euthanized. The ileum was harvested for a histological evaluation and the measurement of the mRNA and protein expression.


The rate of NEC-like intestinal injury in the treatment group (9/25, 36%) was significantly lower than in the NEC group (25/34, 73.5%). Tissue levels of TNF-α, IL-6 and HMGB1 were significantly elevated in the NEC group, whereas those in the treatment group were decreased to similar values as in the control group.


Our experimental study showed that rTM is able to reduce the severity and incidence of NEC. It may be an alternative option for the treatment of NEC.


Necrotizing enterocolitis Rat Cytokines Inflammatory Recombinant human soluble thrombomodulin 


Compliance with ethical standards

Conflict of interest

The authors declare no conflicts of interest in association with this study.

Supplementary material

595_2019_1832_MOESM1_ESM.tif (8 mb)
Supplementary material 1 (TIFF 8209 kb)
595_2019_1832_MOESM2_ESM.tif (8 mb)
Supplementary material 2 (TIFF 8209 kb)


  1. 1.
    Neu J, Walker WA. Necrotizing enterocolitis. N Engl J Med. 2011;364:255–64.CrossRefGoogle Scholar
  2. 2.
    Ganapathy V, Hay JW, Kim JH, Lee ML, Rechtman DJ. Long term healthcare costs of infants who survived neonatal necrotizing enterocolitis: a retrospective longitudinal study among infants enrolled in Texas Medicaid. BMC Pediatr. 2013;13:127.CrossRefGoogle Scholar
  3. 3.
    Lu P, Sodhi CP, Jia H, Shaffiey S, Good M, Branca MF, et al. Animal models of gastrointestinal and liver diseases. Animal models of necrotizing enterocolitis: pathophysiology, translational relevance, and challenges. Am J Physiol Gastrointest Liver Physiol. 2014;306:G917–928.CrossRefGoogle Scholar
  4. 4.
    Tanner SM, Berryhill TF, Ellenburg JL, Jilling T, Cleveland DS, Lorenz RG, et al. Pathogenesis of necrotizing enterocolitis: modeling the innate immune response. Am J Pathol. 2015;185:4–16.CrossRefGoogle Scholar
  5. 5.
    Morser J. Thrombomodulin links coagulation to inflammation and immunity. Curr Drug Targets. 2012;13:421–31.CrossRefGoogle Scholar
  6. 6.
    Maruyama I. Recombinant thrombomodulin and activated protein C in the treatment of disseminated intravascular coagulation. Thromb Haemost. 1999;82:718–21.CrossRefGoogle Scholar
  7. 7.
    Nagato M, Okamoto K, Abe Y, Higure A, Yamaguchi K. Recombinant human soluble thrombomodulin decreases the plasma high-mobility group box-1 protein levels, whereas improving the acute liver injury and survival rates in experimental endotoxemia. Crit Care Med. 2009;37:2181–6.CrossRefGoogle Scholar
  8. 8.
    Kadono K, Uchida Y, Hirao H, Miyauchi T, Watanabe T, Iida T, et al. Thrombomodulin attenuates inflammatory damage due to liver ischemia and reperfusion injury in mice in toll-like receptor 4-dependent manner. Am J Transplant. 2017;17:69–80.CrossRefGoogle Scholar
  9. 9.
    Hunter CJ, De Plaen IG. Inflammatory signaling in NEC: role of NF-κB, cytokines and other inflammatory mediators. Pathophysiology. 2014;21:55–65.CrossRefGoogle Scholar
  10. 10.
    Osumi W, Jin D, Imai Y, Tashiro K, Li ZL, Otsuki Y, et al. Recombinant human soluble thrombomodulin improved lipopolysaccharide/d-galactosamine-induced acute liver failure in mice. J Pharmacol Sci. 2015;129:233–9.CrossRefGoogle Scholar
  11. 11.
    Rajashekhar G, Gupta A, Marin A, Friedrich J, Willuweit A, Berg DT, et al. Soluble thrombomodulin reduces inflammation and prevents microalbuminuria induced by chronic endothelial activation in transgenic mice. Am J Physiol Renal Physiol. 2012;302:703–12.CrossRefGoogle Scholar
  12. 12.
    Barlow B, Santulli TV. Importance of multiple episodes of hypoxia or cold stress on the development of enterocolitis in an animal model. Surgery. 1975;77:687–90.Google Scholar
  13. 13.
    Nakame K, Kaji T, Mukai M, Shinyama S, Matsufuji H. The protective and anti-inflammatory effects of glucagon-like peptide-2 in an experimental rat model of necrotizing enterocolitis. Peptides. 2016;75:1–7.CrossRefGoogle Scholar
  14. 14.
    Feng J, El-Assal ON, Besner GE. Heparin-binding EGF-like growth factor (HB-EGF) and necrotizing enterocolitis. Semin Pediatr Surg. 2005;14:167–74.CrossRefGoogle Scholar
  15. 15.
    Dvorak B, Halpern MD, Holubec H, Williams CS, McWilliam DL, Dominguez JA, et al. Epidermal growth factor reduces the development of necrotizing enterocolitis in a neonatal ratmodel. Am J Physiol Gastrointest Liver Physiol. 2002;282:156–64.CrossRefGoogle Scholar
  16. 16.
    Ares GJ, McElroy SJ, Hunter CJ. The science and necessity of using animal models in the study of necrotizing enterocolitis. Semin Pediatr Surg. 2018;27:29–33.CrossRefGoogle Scholar
  17. 17.
    Rentea RM, Welak SR, Fredrich K, Donohoe D, Pritchard KA, Oldham KT, et al. Early enteral stressors in newborns increase inflammatory cytokine expression in a neonatal necrotizing enterocolitis rat model. Eur J Pediatr Surg. 2013;23:39–47.Google Scholar
  18. 18.
    Ikeguchi H, Maruyama S, Morita Y, Fujita Y, Kato T, Natori Y, et al. Effects of human soluble thrombomodulin on experimental glomerulonephritis. Kidney Int. 2002;61:490–501.CrossRefGoogle Scholar
  19. 19.
    Ito T, Kawahara K, Okamoto K, Yamada S, Yasuda M, Imaizumi H, et al. Proteolytic cleavage of high mobility group box 1 protein by thrombin-thrombomodulincomplexes. Arterioscler Thromb Vasc Biol. 2008;28:1825–30.CrossRefGoogle Scholar
  20. 20.
    Wang H, Yang H, Czura CJ, Sama AE, Tracey KJ. HMGB1 as a late mediator of lethal systemic inflammation. Am J Respir Crit Care Med. 2001;164:1768–73.CrossRefGoogle Scholar
  21. 21.
    Scaffidi P, Misteli T, Bianchi ME. Release of chromatin protein HMGB1 by necrotic cells triggers inflammation. Nature. 2002;418:191–5.CrossRefGoogle Scholar
  22. 22.
    Schmidt AM, Yan SD, Yan SF, Stern DM. The multiligand receptor RAGE as a progression factor amplifying immune and inflammatory responses. J Clin Invest. 2001;108:949–55.CrossRefGoogle Scholar
  23. 23.
    Abeyama K, Stern DM, Ito Y, Kawahara K, Yoshimoto Y, Tanaka M, et al. The N-terminal domain of thrombomodulin sequesters high-mobility group-B1 protein, a novel antiinflammatory mechanism. J Clin Invest. 2005;115:1267–74.CrossRefGoogle Scholar
  24. 24.
    Zamora R, Grishin A, Wong C, Boyle P, Wang J, Hackam D, et al. High-mobility group box 1 protein is an inflammatory mediator in necrotizing enterocolitis: protective effect of the macrophage deactivator semapimod. Am J Physiol Gastrointest Liver Physiol. 2005;289:G643–652.CrossRefGoogle Scholar
  25. 25.
    Feng J, El-Assal ON, Besner GE. Heparin-binding epidermal growth factor-like growth factor decreases the incidence of necrotizing enterocolitis in neonatal rats. J Pediatr Surg. 2006;41:144–9.CrossRefGoogle Scholar
  26. 26.
    Downard CD, Grant SN, Matheson PJ, Guillaume AW, Debski R, Fallat ME, et al. Altered intestinal microcirculation is the critical event in the development of necrotizing enterocolitis. J Pediatr Surg. 2011;46:1023–8.CrossRefGoogle Scholar
  27. 27.
    Zhang HY, Wang F, Feng JX. Intestinal microcirculatory dysfunction and neonatal necrotizing enterocolitis. Chin Med J (Engl). 2013;126:1771–8.Google Scholar
  28. 28.
    Watkins DJ, Besner GE. The role of the intestinal microcirculation in necrotizing enterocolitis. Semin Pediatr Surg. 2013;22:83–7.CrossRefGoogle Scholar

Copyright information

© Springer Nature Singapore Pte Ltd. 2019

Authors and Affiliations

  • Bo Li
    • 1
  • Ryuta Saka
    • 1
  • Yuichi Takama
    • 1
  • Takehisa Ueno
    • 1
  • Yuko Tazuke
    • 1
  • Hiroomi Okuyama
    • 1
    Email author
  1. 1.Department of Pediatric SurgeryOsaka University Graduate School of MedicineSuitaJapan

Personalised recommendations