Anti-HMGB1 neutralizing antibody ameliorates gut barrier dysfunction and improves survival after hemorrhagic shock - PubMed (original) (raw)

Comparative Study

. 2006 Apr-Jun;12(4-6):105-14.

doi: 10.2119/2006-00010.Yang.

Tomoyuki Harada, Kevin P Mollen, Jose M Prince, Ryan M Levy, Joshua A Englert, Margot Gallowitsch-Puerta, LiHong Yang, Huan Yang, Kevin J Tracey, Brian G Harbrecht, Timothy R Billiar, Mitchell P Fink

Affiliations

Comparative Study

Anti-HMGB1 neutralizing antibody ameliorates gut barrier dysfunction and improves survival after hemorrhagic shock

Runkuan Yang et al. Mol Med. 2006 Apr-Jun.

Abstract

Intestinal barrier dysfunction occurs following hemorrhagic shock and resuscitation (HS/R). High-mobility group B1 (HMGB1) has been shown to increase the permeability of Caco-2 human enterocyte-like epithelial monolayers in vitro. In this study, we found that serum concentrations of HMGB1 were higher in blood samples obtained from 25 trauma victims with hemorrhagic shock than in 9 normal volunteers. We also studied whether treatment with anti-HMGB1 antibody can ameliorate HS/R-induced gut barrier dysfunction in mice. Animals were shocked by withdrawal of blood to maintain mean arterial pressure at 25 to 30 mmHg for 2 h. After resuscitation with shed blood plus Ringer's lactate solution, the mice were treated with either anti-HMGB1 antibody or nonimmune rabbit IgG. Serum HMGB1 concentrations were significantly higher in trauma victims than control mice. Treatment with anti-HMGB1 antibody improved survival at 24 h and ameliorated the development of ileal mucosal hyperpermeability to FITC-labeled dextran. At 24 h after HS/R, treatment with anti-HMGB1 antibody decreased bacterial translocation to mesenteric lymph nodes and was associated with lower circulating concentrations of IL-6 and IL-10. These data support the notion that HMGB1 is a mediator of HS/R-induced gut barrier dysfunction and suggest that anti-HMGB1 antibodies warrant further evaluation as a therapeutic to ameliorate the morbidity of HS/R in trauma patients.

PubMed Disclaimer

Figures

Figure 1

Figure 1

Serum HMGB1 concentrations in trauma victims with hemorrhagic shock and healthy human volunteers (n = 9). Blood samples from the trauma victims were obtained as soon as possible after admission to the Emergency Department and always < 6 h after the time of injury (n = 25) and again the next day (i.e., approximately 24 h after injury; n = 24). Serum HMGB1 concentrations in the trauma victims at either time point were significantly greater than the values measured in the healthy subjects (P < 0.0001).

Figure 2

Figure 2

Effect of treatment with anti-HMGB1 antibody on HS/R-induced gut mucosal permeability (A) and bacterial translocation to MLN (B). Ileal mucosal permeability and bacterial translocation were assessed 24 h after HS/R (or the sham procedure). Mice in the sham group (n = 6) were subjected to anesthesia and vascular cannulation but not hemorrhage. Mice in the IgG (n = 6) and aHMGB1 (n = 8) groups were shocked by withdrawal of blood over 10 min until MAP decreased to 25 mmHg. MAP was maintained at 25 to 30 mmHg for 2 h, and then the mice were resuscitated by infusing all of the remaining shed blood plus a volume of Ringer’s lactate solution equal to twice the total volume of shed blood. Mice in the IgG group were treated with a single intraperitoneal dose of rabbit IgG (600 μg in 500 μL) immediately after the resuscitation procedure. Mice in the aHMGB1 group were similarly treated, except these animals received a dose of a polyclonal rabbit neutralizing aHMGB1 antibody. Mice in the sham group were treated with PBS (500 μL intraperitoneally). Results are means ± SEM. *P < 0.05 vs. sham; †P < 0.05 vs. IgG.

Figure 3

Figure 3

Effect of treatment with anti-HMGB1 antibody on HS/R-induced changes in serum IL-6 (A) and IL-10 (B) concentrations. Cytokine levels were assessed 24 h after HS/R (or the sham procedure). Groups and sample sizes are the same as in Figure 1. Results are means ± SEM. *P < 0.05 vs. sham; †P < 0.05 vs. IgG.

Figure 4

Figure 4

Effect of treatment with IgG or HMGB1 antibody on the expression of TNF, iNOS, and IL-6 mRNA in samples of intestinal mucosal tissue. RT-PCR was performed using tissue samples obtained 24 h after HS/R. Bands visualized after agarose gel electrophoresis of PCR reaction products were scanned using a NucleoVision imaging workstation and quantified using GelExpert release 3.5. Data are means ± SE (n = 3 per condition). A typical gel is depicted.

Figure 5

Figure 5

Effect of treatment with anti-HMGB1 antibody on HS/R-induced changes in ZO-1, occludin, and RAGE protein expression in gut mucosal tissue lysates. Western blots were performed using mucosal extracts prepared from tissues obtained 24 h after HS/R or the sham procedure. The groups are same as in Figure 4. The figure depicts results from representative assays that were performed 3 or 4 times with comparable findings. Typical gels are depicted.

Figure 6

Figure 6

Effect of treatment with anti-HMGB1 antibody on HS/R-induced changes in serum ALT concentration assessed 24 h after HS/R (or the sham procedure). Groups and sample sizes are the same as in Figure 1. Results are means ± SEM. *P < 0.05 vs. sham; †P < 0.05 vs. IgG.

Similar articles

Cited by

References

    1. Anderson RN, Smith BL. Deaths: Leading causes for 2002. National Vital Statistics Reports. 2005;53:1–90. - PubMed
    1. Sauaia A, et al. Epidemiology of trauma deaths: a reassessment. J Trauma. 1995;38:185–93. - PubMed
    1. Acosta JA, et al. Lethal injuries and time to death in a level I trauma center. J Am Coll Surg. 1998; 186:528–33. - PubMed
    1. Sauaia A, Moore FA, Moore EE, Norris JM, Lezotte DC, Hamman RF. Multiple organ failure can be predicted as early as 12 hours after injury. J Trauma. 1998;45:301–3. - PubMed
    1. Moore FA, Moore EE, Sauaia A. Blood transfusion: an independent risk factor for postinjury multiple organ failure. Arch Surg. 1997;132:620–4. - PubMed

Publication types

MeSH terms

Substances

Grants and funding

LinkOut - more resources