Critical Care Medicine  |   October 2019
Recombinant Thrombomodulin on Neutrophil Extracellular Traps in Murine Intestinal Ischemia–Reperfusion
Author Notes
  • From the Departments of Acute Medicine (N.H., K.D., T.H., N.M.) and Nephrology and Endocrinology (R.M., Y.H., E.N., M.N.), University of Tokyo, Tokyo, Japan.
  • This article is featured in “This Month in Anesthesiology,” page 1A.
    This article is featured in “This Month in Anesthesiology,” page 1A.×
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    Supplemental Digital Content is available for this article. Direct URL citations appear in the printed text and are available in both the HTML and PDF versions of this article. Links to the digital files are provided in the HTML text of this article on the Journal’s Web site (www.anesthesiology.org).×
  • Part of the work presented in this article was presented at the 30th Annual Congress of the European Society of Intensive Care Medicine on September 26, 2017, in Vienna, Austria.
    Part of the work presented in this article was presented at the 30th Annual Congress of the European Society of Intensive Care Medicine on September 26, 2017, in Vienna, Austria.×
  • Submitted for publication January 28, 2019. Accepted for publication June 17, 2019.
    Submitted for publication January 28, 2019. Accepted for publication June 17, 2019.×
  • Address correspondence to Dr. Doi: University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan. kdoi-tky@umin.ac.jp. Information on purchasing reprints may be found at www.anesthesiology.org or on the masthead page at the beginning of this issue. Anesthesiology’s articles are made freely accessible to all readers, for personal use only, 6 months from the cover date of the issue.
Article Information
Critical Care Medicine / Basic Science / Cardiovascular Anesthesia / Critical Care / Gastrointestinal and Hepatic Systems
Critical Care Medicine   |   October 2019
Recombinant Thrombomodulin on Neutrophil Extracellular Traps in Murine Intestinal Ischemia–Reperfusion
Anesthesiology 10 2019, Vol.131, 866-882. doi:10.1097/ALN.0000000000002898
Anesthesiology 10 2019, Vol.131, 866-882. doi:10.1097/ALN.0000000000002898
Abstract

Editor’s Perspective:

What We Already Know about This Topic:

  • Intestinal ischemia–reperfusion causes multiple-organ dysfunction syndrome

  • Neutrophil extracellular traps and histone activation may be important in the development of multiple-organ dysfunction

  • Recombinant thrombomodulin binds to circulating histones and may modulate their activation

What This Article Tells Us That Is New:

  • In male mice, recombinant thrombomodulin administration after intestinal ischemia–reperfusion increased survival rate

  • Histones and neutrophil extracellular traps were found in the livers and intestines after ischemia–reperfusion, and recombinant thrombomodulin reduced these accumulations only in the liver

  • Recombinant thrombomodulin may attenuate liver injury by suppressing hepatic histone and neutrophil extracellular trap accumulation

Background: In multiple-organ dysfunction, an injury affecting one organ remotely impacts others, and the injured organs synergistically worsen outcomes. Recently, several mediators, including extracellular histones and neutrophil extracellular traps, were identified as contributors to distant organ damage. This study aimed to elucidate whether these mediators play a crucial role in remote organ damage induced by intestinal ischemia–reperfusion. This study also aimed to evaluate the protective effects of recombinant thrombomodulin, which has been reported to neutralize extracellular histones, on multiple-organ dysfunction after intestinal ischemia–reperfusion.

Methods: Intestinal ischemia was induced in male C57BL/6J mice via clamping of the superior mesenteric artery. Recombinant thrombomodulin (10 mg/kg) was administered intraperitoneally with the initiation of reperfusion. The mice were subjected to a survival analysis, histologic injury scoring, quantitative polymerase chain reaction analysis of tumor necrosis factor-α and keratinocyte-derived chemokine expression, Evans blue dye vascular permeability assay, and enzyme-linked immunosorbent assay analysis of histones in the jejunum, liver, lung, and kidney after 30- or 45-min ischemia. Neutrophil extracellular trap formation was evaluated by immunofluorescence staining.

Results: Recombinant thrombomodulin yielded statistically significant improvements in survival after 45-min ischemia (ischemia–reperfusion without vs. with 10 mg/kg recombinant thrombomodulin: 0% vs. 33%, n = 21 per group, P = 0.001). Recombinant thrombomodulin reduced the histologic injury score, expression of tumor necrosis factor-α and keratinocyte-derived chemokine, and extravasation of Evans blue dye, which were augmented by 30-min ischemia–reperfusion, in the liver, but not in the intestine. Accumulated histones and neutrophil extracellular traps were found in the livers and intestines of 30-min ischemia–reperfusion–injured mice. Recombinant thrombomodulin reduced these accumulations only in the liver.

Conclusions: Recombinant thrombomodulin improved the survival of male mice with intestinal ischemia–reperfusion injury. These findings suggest that histone and neutrophil extracellular trap accumulation exacerbate remote liver injury after intestinal ischemia–reperfusion. Recombinant thrombomodulin may suppress these accumulations and attenuate liver injury.