Recombinant factor XIII mitigates hemorrhagic shock-induced organ dysfunction

Abstract

Background: Plasma factor XIII (FXIII) is responsible for stabilization of fibrin clot at the final stage of blood coagulation. Since FXIII has also been shown to modulate inflammation, endothelial permeability, as well as diminish multiple organ dysfunction (MOD) after gut ischemia-reperfusion injury, we hypothesized that FXIII would reduce MOD caused by trauma-hemorrhagic shock (THS).

Materials and methods: Rats were subjected to a 90 min THS or trauma sham shock (TSS) and treated with either recombinant human FXIII A(2) subunit (rFXIII) or placebo immediately after resuscitation with shed blood or at the end of the TSS period. Lung permeability, lung and gut myeloperoxidase (MPO) activity, gut histology, neutrophil respiratory burst, microvascular blood flow in the liver and muscles, and cytokine levels were measured 3 h after the THS or TSS. FXIII levels were measured before THS or TSS and after the 3-h post-shock period.

Results: THS-induced lung permeability as well as lung and gut MPO activity was significantly lower in rFXIII-treated than in placebo-treated animals. Similarly, rFXIII-treated rats had lower neutrophil respiratory burst activity and less ileal mucosal injury. rFXIII-treated rats also had a higher liver microvascular blood flow compared with the placebo group. Cytokine response was more favorable in rFXIII-treated animals. Trauma-hemorrhagic shock did not cause a drop in FXIII activity during the study period.

Conclusions: Administration of rFXIII diminishes THS-induced MOD in rats, presumably by preservation of the gut barrier function, limitation of polymorphonuclear leukocyte (PMN) activation, and modulation of the cytokine response.

FIG. 1

rFXIII reduces the severity of ileal mucosal damage after THS. Hematoxylin-eosin staining. Magnification ×100. (A) Rat is subjected to THS and placebo treatment. Villi tips are denuded (arrows). Red blood cell congestion is seen. Gut injury level is classified as grade 3 mucosal damage score. (B) Rat is subjected to THS and rFXIII treatment. Extension of the subepithelial space as well as villi edema (arrows) is seen. Gut injury level is classified as grade 2 mucosal damage score. (C) Rat is subjected to TSS and placebo treatment. Almost normal villi are seen (grades 0–1 mucosal damage score). (D) Rat is subjected to TSS and rFXIII treatment. Almost normal villi are seen (grades 0–1 mucosal damage score).

FIG. 2

rFXIII diminishes lung permeability alterations after THS. Lung permeability is evaluated by measuring concentration of EBD in bronchoalveolar lavage fluid and expressed as the percentage of that present in the plasma. Data are expressed as means ± SD (n = 8 in each group). *P < 0.05 versus all other groups; **P < 0.05 versus both shams.

FIG. 3

rFXIII reduces neutrophil sequestration in lung and gut after THS. The level of neutrophil sequestration is evaluated by measuring myeloperoxidase activity. Data are expressed as means ± SD (n = 8 in each group). *P < 0.01 versus all other groups. **P < 0.01 versus both shams.

FIG. 4

rFXIII decreases the level of oxidative stress after THS. The level of oxidative stress is assessed by neutrophil respiratory burst activity. Data are expressed as means ± SD (n = 8 in each group). *P < 0.05 versus all other groups; **P < 0.05 versus both shams.

FIG. 5

THS does not decrease FXIII activity. FXIII activity in rat plasma is determined using Berichrom assay before and 3 h after THS or TSS. Data are expressed as means ± SD (n = 8 in each group). *P < 0.05 versus before THS/TSS; **P < 0.05 versus groups with placebo treatment.