Share this article on:

The Fibrin-Derived Peptide Bβ15-42 Attenuates Liver Damage in a Rat Model of Liver Ischemia/Reperfusion Injury

Liu, Anding*†; Fang, Haoshu; Yang, Yan*; Sun, Jian; Fan, Hua§; Liu, Shenpei*; Dirsch, Olaf; Dahmen, Uta

doi: 10.1097/SHK.0b013e31828c2b75
Basic Science Aspects

ABSTRACT The inflammatory response after liver ischemia/reperfusion (I/R) contributes to increased risk of liver failure after liver surgery. Strategies aimed to preventing inflammation could be beneficial in reducing liver I/R injury. Recent studies have demonstrated that peptide Bβ15-42 is able to decrease the injury of I/R in heart and kidney by inhibition of leukocyte migration and preserving endothelial barrier function. Prompted by these results, we hypothesized that Bβ15-42 could also possess anti-inflammatory abilities to protect from or reduce hepatic I/R injury. Therefore, in this study, we aimed to evaluate the effects of Bβ15-42 in a model of liver I/R injury in rats. Rats were treated with Bβ15-42 at initiation of reperfusion and 2 h thereafter. Rats were killed at 0.5, 6, 24, and 48 h after reperfusion. Hepatic mRNA levels of fibrinogen-α (Fgα), Fgβ, Fgγ were significantly increased after I/R. Treatment with Fg-derived Bβ15-42 ameliorated liver I/R injury, as indicated by lower serum aminotransferase levels and fewer I/R-associated histopathologic changes. Bβ15-42 treatment decreased leukocyte infiltration and expression of hepatic inflammatory cytokines. Moreover, Bβ15-42 significantly reduced high-mobility group box 1 release and altered mitogen-activated protein kinase activation. In conclusion, Bβ15-42 treatment protected against liver warm I/R injury. The mechanism of protective action of Bβ15-42 seemed to involve its ability to reduce hepatic inflammatory response through preventing high-mobility group box 1 release and altering mitogen-activated protein kinase activation.

*Experimental Medicine Center, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China; Experimental Transplantation Surgery, Department of General, Visceral and Vascular Surgery, Friedrich-Schiller-University Jena, Jena, Germany; Department of General Surgery, Sun Yat-sen Memorial Hospital of Sun Yat-sen University, Guangzhou; and §Department of Hepatobiliary Surgery, Beijing Chaoyang Hospital, Capital Medical University, Beijing, China; and Institute for Pathology, Friedrich-Schiller-University Jena, Jena, Germany.

Received 24 Nov 2012; first review completed 17 Dec 2012; accepted in final form 7 Feb 2013

Address reprint requests to Anding Liu, MD, Experimental Medicine Center, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, No. 1095 Jiefang Ave, Wuhan 430030, China. E-mail: Anding.liu@uk-essen.de.

The authors declare no conflicts of interest.

Back to Top | Article Outline

INTRODUCTION

Ischemia/reperfusion (I/R) injury is triggered when the liver is transiently deprived of oxygen and subsequently reoxygenated. Injury to the liver caused by I/R can occur in the procedure of liver transplantation, liver resection, and trauma (1–3). Liver I/R injury is implicated as a potent contributor in increasing the rate of acute liver failure and chronic liver dysfunction after liver surgery, therefore, the reduction of liver I/R injury can contribute to minimizing postoperative injury to the liver.

The mechanisms of liver damage in I/R is associated with leukocyte infiltration and subsequent release of cytokines, free radicals, and proteolytic enzymes (4, 5). Several studies have demonstrated that blocking post-I/R leukocyte recruitment protects against liver I/R injury (6–8). The process of leukocyte recruitment across endothelial and extracellular matrix barriers involves complex cascades of tethering, rolling, adhesion to the endothelium, and transmigration from the vasculature. Leukocyte adhesion is mediated through interaction with adhesive molecules, such as L-selectin, intercellular adhesion molecule 1, and vascular cell adhesion molecule 1. Vascular endothelial (VE) cadherin is one of the key molecules of mediating endothelial cell-cell contacts at intercellular junctions (9). Vascular endothelial cadherin has binding sites for a part of the amino terminus of the β-chain of fibrin. Binding VE cadherin to the β-chain of fibrin leads to transmigration of leukocytes across endothelial cell monolayers (10, 11).

The peptide Bβ15-42 is a 28–amino acid corresponding to the N-terminal sequence of the β-chain of fibrin. Because of lack of a leukocyte binding site, Bβ15-42 competes with fibrin for binding to VE cadherin, thereby inhibiting leukocyte transmigration across endothelial junctions and reducing the inflammatory response (11–13). Recently, results from Groger et al. (12) showed that Bβ15-42 prevents stress-induced RhoA activation, leading to endothelial stabilization and preservation of the endothelial barrier in shock.

It has been shown that Bβ15-42 peptide reduces myocardial inflammation and infarct size in rodent and pig models of myocardial I/R (13–15), and the results have been transferred successfully in a subsequent clinical phase II trial in patients with acute myocardial infarction undergoing primary percutaneous coronary intervention (16). The protective effect of Bβ15-42 could be further confirmed in kidney I/R injury, hemorrhagic shock and reperfusion, lipopolysaccaride-induced shock, and sepsis animal models (12, 17–20).

Prompted by these results, we hypothesized that Bβ15-42 could also possess anti-inflammatory abilities to protect from or reduce hepatic I/R injury, which is characterized as an inflammatory injury. To test this hypothesis, we investigated the effect of Bβ15-42 in a rat model of liver I/R.

Back to Top | Article Outline

MATERIALS AND METHODS

Experimental design

The experiments were designed to investigate whether Bβ15-42 treatment could protect from liver I/R injury. Rats were treated with Bβ15-42 (2.4 mg/kg) or control random peptide (2.4 mg/kg) at initiation of reperfusion, followed by a second injection 2 h after the onset of reperfusion. The dose of 2.4 mg/kg Bβ15-42 was based on dose titrations in rodent models of myocardial I/R injury (13, 15). Rats were killed at 0.5, 6, 24, and 48 h of reperfusion. Liver injury, inflammatory cytokines, neutrophil infiltration, hepatic high-mobility group box 1 (HMGB1) expression and release, and mitogen-activated protein kinase (MAPK) activation were analyzed.

Back to Top | Article Outline

Synthesis of peptides

Endotoxin-free Bβ15-42 (GHRPLDKKREEAPSLRPAPPPISGGGYR) control random peptide (DRGAPAHRPPRGPISGRSTPEKEKLLPG) were produced by solid-phase peptide synthesis and purified with reverse-phase HPLC as described previously (13).

Back to Top | Article Outline

Animals

Male inbred Lewis rats, purchased from Vital River Cooperation (Beijing, China), weighing within 270 to 320 g, were used in this study. All animals were housed under standard animal care conditions and had free to access to water and rat chow ad libitum. All procedures were carried out according to the ethical guidelines of the Animal Care and Use Committee of Huazhong University of Science and Technology.

Back to Top | Article Outline

Partial hepatic warm I/R

Partial hepatic warm I/R model was performed as described previously (21). In brief, after opening the abdomen and dissecting interlobular ligaments, a microvascular clamp was used to interrupt the arterial and portal venous blood supply to the left lateral and median liver lobes for 60 min.

Back to Top | Article Outline

Histopathology

Liver tissue was fixed in 4.5% buffered formalin for at least 24 h. Paraffin embedding was performed using standard techniques. Sections (4 μm) were stained with hematoxylin-eosin and assessed for inflammation and tissue damage.

Back to Top | Article Outline

Naphthol-AS-D-chloroacetate esterase staining

Neutrophil infiltration into liver tissue was evaluated by naphthol-AS-D-chloroacetate esterase (ASDCL) staining, as reported previously (22–24). Each sample was evaluated for ASDCL staining–positive neutrophils per high-power field (HPF) of five random sections with magnification of 200×. Naphthol-AS-D-chloroacetate esterase staining–positive neutrophils were counted manually.

Back to Top | Article Outline

Gel electrophoresis and Western blotting

Western blotting was performed as described previously (25). In brief, proteins were separated by sodium dodecyl sulfate–polyacrylamide gel electrophoresis and transferred onto polyvinyldifluoride membranes. Antibodies against rabbit anti-HMGB1 (1:1,000; Abcam, Cambridge, UK), rabbit anti–phosphor–extracellular signal–regulated kinase (ERK) (Thr202/Tyr204, 1:1,000; Cell Signaling Technology, Beverly, Mass), rabbit anti-ERK (1:1,000; Cell Signaling Technology), rabbit anti–phosphor–c-Jun N-terminal kinase (JNK) (Thr183/Tyr185, 1:1,000; Cell Signaling Technology), rabbit anti-JNK (1:1,000; Cell Signaling Technology), rabbit anti–phospho-p38 (Thr180/Tyr182, 1:1,000; Cell Signaling Technology), rabbit anti-p38 (1:1,000; Cell Signaling Technology), and rabbit anti–glyceraldehyde 3-phosphate dehydrogenase (1:20,000; Sigma-Aldrich, St. Louis, Mo) were used for Western blot analysis. The gray value of bands was calculated by ImageJ 1.43 G (National Institutes of Health, Bethesda, Md).

Back to Top | Article Outline

Enzyme-linked immunosorbent assay

Serum HMGB1 was determined with a commercial enzyme-linked immunosorbent assay ([ELISA] Shino-Test, Kanagawa, Japan) according to the manufacturer’s instruction. Serum tumor necrosis factor-α (TNF-α) and interleukin 6 (IL-6) levels were analyzed using commercially available ELISA kits (R&D Systems, Minneapolis, Minn).

Back to Top | Article Outline

Quantitative polymerase chain reaction

Total RNA was isolated by TRIzol Reagent (Invitrogen, Carlsbad, Calif) according to the manufacturer’s instruction. Complementary DNA synthesis was performed using the First-Strand cDNA synthesis kit (Invitrogen). The levels of mRNA expression were determined by real-time polymerase chain reaction (PCR) using a Roch Light cycle system (Roche, Rotkreuz, Switzerland) with SYBR green master mix (Qiagen, Hilden, Germany) and primers (Table 1). Thermal cycling conditions consisted of a 10-min template denaturation step at 95°C, followed by 50 cycles of 95°C for 30 s, 50 °C for 30 s and 72 °C for 20 s. Normal liver tissue was used as reference sample to generate the standard curve. Relative quantification of target mRNA expression was calculated and further normalized to hypoxanthine guanine phosphoribosyltransferase.

Table 1

Table 1

Back to Top | Article Outline

Statistical analysis

Data were expressed as mean ± SD. Differences between groups were evaluated for significance by one-way analysis of variance combined with Bonferroni post hoc test. A value of P < 0.05 was considered to indicate statistical significance.

Back to Top | Article Outline

RESULTS

Fgα, Fgβ, Fgγ mRNA expression is increased in the liver after warm I/R injury

It has been reported that expression of Fg is increased in a rat model of kidney I/R (20). To determine whether liver warm I/R injury also could upregulate Fg expression, hepatic mRNA levels for Fgα, Fgβ, and Fgγ were measured by quantitative PCR. As shown in Figure 1, after 60 min of warm ischemia, hepatic Fgα, Fgβ, and Fgγ mRNA expression levels were significantly increased as early as 6 h, reached a peak at 24 h, and remained higher at 48 h after reperfusion, accounting for a 5-, 10-, 5-fold increase at 24 h of reperfusion, respectively.

Fig. 1

Fig. 1

Back to Top | Article Outline

Bβ15-42 treatment protects against liver I/R injury

Bβ15-42 has been demonstrated to protect against inflammation and injury after I/R in heart and kidney (13–15, 17, 20, 26). To determine whether Bβ15-42 treatment could attenuate liver I/R injury, a total dose of Bβ15-42 or control random peptide was administered to rats at initiation of reperfusion and 2 h thereafter. As shown in Figure 2A, liver warm I/R resulted in significant increases in serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST). After 60 min of warm ischemia injury, serum ALT was significantly increased at 6 and 24 h of reperfusion (6 h, 890 ± 119 IU/L; 24 h, 329 ± 75 IU/L vs. normal control, 58 ± 7 IU/L; P < 0.001). In contrast, Bβ15-42 treatment significantly decreased ALT and AST levels when compared with random peptide control group. The serum levels of ALT were drastically decreased after reperfusion to 487 ± 85 IU/L and 201 ± 55 IU/L at 6 and 24 h, respectively. Similar results were obtained for AST.

Fig. 2

Fig. 2

As shown in Figure 2B, liver histology confirmed the serum aminotransferase estimation of liver injury. Hepatocellular necrosis and cytoplasmic vacuolization of hepatocytes were observed 6 h after reperfusion in the control group. In contrast, minimal damage was noted in Bβ15-42–treated rats.

Back to Top | Article Outline

Bβ15-42 treatment decreases the production of inflammatory mediators

Hepatic TNF-α, IL-6, IL-1β, and inducible nitric oxide synthase (iNOS) mRNA expression levels were measured by quantitative PCR. As shown in Figure 3, A to D, after 60 min of warm ischemia and 6 h of reperfusion, hepatic TNF-α, IL-6, IL-1β, and iNOS mRNA expression levels were significantly increased in both random peptide control group and Bβ15-42 treatment group. However, Bβ15-42–injected rats expressed significantly less hepatic TNF-α, IL-6, IL-1β, and iNOS mRNA compared with random peptide-treated rats. Furthermore, serum TNF-α and IL-6 levels in Bβ15-42–treated rats were significantly less than those in control random peptide–treated rats (Fig. 3, E and F).

Fig. 3

Fig. 3

Back to Top | Article Outline

Bβ15-42 decreases hepatic neutrophil infiltration

For quantification of the inflammatory response in livers subjected to 60 min of warm ischemia and 24 h of reperfusion, the number of ASDCL staining–positive neutrophils infiltrating the liver was determined. The number of ASDCL staining–positive neutrophils was increased after I/R injury. Neutrophil infiltration was significantly less in Bβ15-42–treated rats than in control random peptide–injected rats, indicating a profound abrogation of leukocyte infiltration into the liver (Fig. 4).

Fig. 4

Fig. 4

Back to Top | Article Outline

Bβ15-42 treatment decreases HMGB1 translocation and release

Warm liver I/R injury resulted in upregulation of HMGB1 expression and release in hepatic I/R (21, 27). As shown in Figure 5, I/R increased hepatic HMGB1 expression when compared with normal rats. Of note, Bβ15-42–treated rats exhibited a minimal increase in HMGB1 protein compared with random peptide–injected rats. To assess the extracellular release of HMGB1 after warm liver I/R injury, serum HMGB1 concentration was quantified using an ELISA. We previously demonstrated that serum levels of HMGB1 were highest at 0.5 h after reperfusion and rapidly decreased thereafter (21). Therefore, we determined the serum HMGB1 levels at 0.5 h after reperfusion. Serum HMGB1 levels were significantly increased in both groups when compared with normal rats. However, Bβ15-42–treated rats showed lower serum HMGB1 levels when compared with control random peptide–injected rats (10.7 ± 3.1 ng/mL vs. 5.9 ± 2.0 ng/mL; P < 0.05).

Fig. 5

Fig. 5

Back to Top | Article Outline

Bβ15-42 treatment modulates liver I/R–induced inflammatory signaling pathways

Among the most proximal events in I/R, the best known is the activation of mitogen-activated protein kinases (MAPKs) (28). To determine if Bβ15-42 treatment could influence MAPK activation, we assessed phosphorylation of ERK, JNK, and p38. After I/R, phosphorylation of ERK was increased at 0.5 h after reperfusion, and then decreased thereafter. However, phosphorylations of ERK levels were higher in Bβ15-42–treated rats than that in control random peptide–treated rats. In addition, liver I/R–induced JNK and p38 activation was decreased in Bβ15-42–treated rats. Treatment with Bβ15-42 did not affect total cellular levels of ERK, JNK, and p38. Thus, these data suggest that Bβ15-42 treatment could modulate liver I/R–induced inflammatory signaling pathways (Fig. 6).

Fig. 6

Fig. 6

Back to Top | Article Outline

DISCUSSION

Hepatic I/R related to leukocyte-dependent inflammatory response remains a significant complication in liver surgery. Recent studies suggest that Bβ15-42 has protective properties in I/R injury in heart and kidney (13–15, 17, 20, 26). However, its role in the pathogenesis of liver I/R injury has not been explored. In the present study, we documented that hepatic Fgα, Fgβ, and Fgγ mRNA expression levels were increased after liver I/R injury. Furthermore, we demonstrated that rats treated with Fgβ-derived Bβ15-42 peptide presented reduced liver enzymes, reduced hepatic inflammatory cytokine levels, and neutrophil infiltration. The protective mechanism of Bβ15-42 on hepatic I/R injury seems to involve its ability to reduce HMGB1 release and modulate liver I/R–induced MAPK activation.

Fibrinogen, a major blood plasma clotting protein, is made up of two sets of three different polypeptide chains (Fgα, Fgβ, and Fgγ). In addition to its role in blood coagulation, Fg has been recognized as an important regulator of tissue repair. Drew et al. (29) demonstrated that Fg seems to be important in tissue repair, and Fg-deficient mice showed a delayed wound closure and reduced tensile strength. Fibrinogen has also been shown to regulate the tissue repair process in neurons and lung epithelial cells (30). Furthermore, Krishnamoorthy et al. (20) found that Fg was increased in the kidney as a protective mechanism to facilitate cell proliferation and tissue repair in kidney I/R injury. Here, we showed that hepatic Fg was upregulated, suggesting that Fg may function as a protective mechanism after hepatic I/R injury.

Currently, there is only indirect evidence for the role of Fg-derived products in liver I/R injury. For example, Harada et al. (31) demonstrated that serum levels of fragment E of fibrin and Fg degradation products were increased after liver I/R injury in rats. In fact, recent studies have shown a pathogenic role for Fg-derived products in other organ systems. Fibrinogen-β–derived Bβ15-42 peptide presented reduced organ injury by inhibition of infiltrating leukocytes in I/R in heart and kidney (13–15, 17, 26). We showed that Bβ15-42 administration substantially reduced inflammation and injury in liver after I/R. These findings provide direct evidence that fibrin(ogen)-derived Bβ15-42 are involved in the pathogenesis of liver I/R injury.

The mechanism of protective action of Bβ15-42 involves its ability to inhibit leukocyte-endothelial interaction, which plays an important role in I/R injury. By binding to VE cadherin, Bβ15-42 blocks potential binding sites for E fragments, which is derived from the N-terminal segments of fibrin. Fragment E carries a binding site for VE cadherin and also CD11c and can function as a bridge molecule between endothelial cells and leukocytes (32). Bβ15-42 competes for the VE cadherin binding site, thereby preventing transmigration of leukocytes across endothelial cell monolayers (13). Furthermore, Groger et al. (12) demonstrated that the molecular key for the protective effect of Bβ15-42 is the src kinase Fyn, which associates with VE cadherin–containing junctions. Bβ15-42 binding to VE cadherin leads to the dissociation of Fyn from VE cadherin. Fyn association with p190RhoGAP results in the inactivation of RhoA, causing endothelial stabilization and preservation of the endothelial barrier in shock (12). More recently, Krishnamoorthy et al. (20) documented that Bβ15-42 administration protected mice from I/R-induced kidney injury by aiding in epithelial cell proliferation and tissue repair. It should be noted that the peptide Bβ15-42 is a natural degradation product occurring after fibrin inactivation. Under normal physiological conditions, Bβ15-42 does not inhibit fibrin fragment interactions with VE cadherin because of the low affinity. When Bβ15-42 was given at a supraphysiological dose, it protected against I/R injury in the heart and kidney (13–15, 20). Consistent with these observations, we found that pharmacologic blocking of interaction by Bβ15-42 reduced leukocyte infiltration and liver I/R injury. To study the effects of Bβ15-42 on proinflammatory cytokine gene expression, rats were preconditioned with Bβ15-42 after reperfusion. We demonstrated that after 60 min of warm ischemia, Bβ15-42 treatment reduced TNF-α, IL-6, IL-1β, and iNOS gene expression after reperfusion. These cytokines are known to play a pivotal role in the pathophysiology of hepatic I/R injury.

Because recent studies suggest that extracellular HMGB1, released from damaged liver cells or secreted by immune cells, causes a proinflammatory intrahepatic microenvironment in the postischemic liver (21, 27), we investigated HMGB1 expression and release. High-mobility group box 1 was initially defined as a nuclear protein that loosely binds to chromatin and plays a pivotal role for bending DNA and regulating transcription (33–35). It has become apparent in recent years that HMGB1 mediates a response to inflammation and tissue damage (35, 36). High-mobility group box 1 in the normal rat liver is mainly present in the nucleus of hepatocytes, and it is translocated from nucleus to cytoplasm and release into circulation in liver I/R injury (21, 25). Furthermore, blocking HMGB1 using a neutralizing antibody to HMGB1 protected against liver I/R injury (27). Our results showed that Bβ15-42 treatment led to a lower hepatic HMGB1 expression and serum levels, suggesting that the mechanism of protective action of Bβ15-42 seems to involve its ability to reduce I/R-induced HMGB1 expression and release.

Because Bβ15-42 treatment led to lower serum HMGB1 levels, which could modulate liver I/R–induced MAPK activation (27), we next examined MAPK activation. Activation of MAPK contributes to the inflammatory response after hepatic I/R (28). Activation of ERK occurred early in ischemic livers, and ERK activation after redox stress is protective in hepatocytes (37). Our results showed that Bβ15-42 pretreatment partially increased I/R-induced ERK1/2 activation when compared with control random peptide–injected rats. Furthermore, we demonstrated that Bβ15-42 decreased I/R-induced JNK and p38 activation. Conversely, JNK and p38 MAPK are phosphorylated and activated after reperfusion, and their activation is associated with the induction of both necrosis and apoptosis in liver I/R. Inhibition of JNK or p38 MAPK activation could decrease hepatic I/R injury in the liver (38, 39). Taken together, these data suggest that modulating MAPK activation is also involved in the hepatic protective effects of Bβ15-42. It remains unclear whether Bβ15-42 could directly alter MAPK activation or whether this is secondary to the Bβ15-42–mediated less inflammatory mediator release, such as HMGB1, which could modulate MAPK activation in liver I/R injury.

In summary, we documented that Bβ15-42 treatment could ameliorate liver I/R injury. The mechanism of protective action of Bβ15-42 seemed to involve its ability to reduce the hepatic inflammatory response by preventing HMGB1 release and altering MAPK activation. Therefore, the protective effect of Bβ15-42 on hepatic warm I/R injury might extend its potential clinical applications in liver surgery.

Back to Top | Article Outline

REFERENCES

1. Fondevila C, Busuttil RW, Kupiec-Weglinski JW: Hepatic ischemia/reperfusion injury—a fresh look. Exp Mol Pathol 74 (2): 86–93, 2003.
2. Selzner N, Rudiger H, Graf R, Clavien PA: Protective strategies against ischemic injury of the liver. Gastroenterology 125 (3): 917–936, 2003.
3. de Rougemont O, Lehmann K, Clavien PA: Preconditioning, organ preservation, and postconditioning to prevent ischemia-reperfusion injury to the liver. Liver Transpl 15 (10): 1172–1182, 2009.
4. Vardanian AJ, Busuttil RW, Kupiec-Weglinski JW: Molecular mediators of liver ischemia and reperfusion injury: a brief review. Mol Med 14 (5–6): 337–345, 2008.
5. Montalvo-Jave EE, Escalante-Tattersfield T, Ortega-Salgado JA, Pina E, Geller DA: Factors in the pathophysiology of the liver ischemia-reperfusion injury. J Surg Res 147 (1): 153–159, 2008.
6. Dulkanchainun TS, Goss JA, Imagawa DK, Shaw GD, Anselmo DM, Kaldas F, Wang T, Zhao D, Busuttil AA, Kato H, et al.: Reduction of hepatic ischemia/reperfusion injury by a soluble P-selectin glycoprotein ligand-1. Ann Surg 227 (6): 832–840, 1998.
7. Teoh NC, Ito Y, Field J, Bethea NW, Amr D, McCuskey MK, McCuskey RS, Farrell GC, Allison AC: Diannexin, a novel annexin V homodimer, provides prolonged protection against hepatic ischemia-reperfusion injury in mice. Gastroenterology 133 (2): 632–646, 2007.
8. Hamada T, Fondevila C, Busuttil RW, Coito AJ: Metalloproteinase-9 deficiency protects against hepatic ischemia/reperfusion injury. Hepatology 47 (1): 186–198, 2008.
9. Vestweber D: VE-cadherin: the major endothelial adhesion molecule controlling cellular junctions and blood vessel formation. Arterioscler Thromb Vasc Biol 28 (2): 223–232, 2008.
10. Gorlatov S, Medved L: Interaction of fibrin(ogen) with the endothelial cell receptor VE-cadherin: mapping of the receptor-binding site in the NH2-terminal portions of the fibrin beta chains. Biochemistry 41 (12): 4107–4116, 2002.
11. Zacharowski K, Zacharowski P, Reingruber S, Petzelbauer P: Fibrin(ogen) and its fragments in the pathophysiology and treatment of myocardial infarction. J Mol Med (Berl) 84 (6): 469–477, 2006.
12. Groger M, Pasteiner W, Ignatyev G, Matt U, Knapp S, Atrasheuskaya A, Bukin E, Friedl P, Zinkl D, Hofer-Warbinek R, et al.: Peptide Bbeta(15-42) preserves endothelial barrier function in shock. PLoS One 4 (4): e5391, 2009.
13. Petzelbauer P, Zacharowski PA, Miyazaki Y, Friedl P, Wickenhauser G, Castellino FJ, Groger M, Wolff K, Zacharowski K: The fibrin-derived peptide Bbeta15-42 protects the myocardium against ischemia-reperfusion injury. Nat Med 11 (3): 298–304, 2005.
14. Roesner JP, Petzelbauer P, Koch A, Mersmann J, Zacharowski PA, Boehm O, Reingruber S, Pasteiner W, Mascher D, Wolzt M, et al.: The fibrin-derived peptide Bbeta15-42 is cardioprotective in a pig model of myocardial ischemia-reperfusion injury. Crit Care Med 35 (7): 1730–1735, 2007.
15. Zacharowski K, Zacharowski PA, Friedl P, Mastan P, Koch A, Boehm O, Rother RP, Reingruber S, Henning R, Emeis JJ, et al.: The effects of the fibrin-derived peptide Bbeta(15-42) in acute and chronic rodent models of myocardial ischemia-reperfusion. Shock 27 (6): 631–637, 2007.
16. Atar D, Petzelbauer P, Schwitter J, Huber K, Rensing B, Kasprzak JD, Butter C, Grip L, Hansen PR, Suselbeck T, et al.: Effect of intravenous FX06 as an adjunct to primary percutaneous coronary intervention for acute ST-segment elevation myocardial infarction results of the F.I.R.E. (Efficacy of FX06 in the Prevention of Myocardial Reperfusion Injury) trial. J Am Coll Cardiol 53 (8): 720–729, 2009.
17. Sorensen I, Rong S, Susnik N, Gueler F, Shushakova N, Albrecht M, Dittrich AM, von Vietinghoff S, Becker JU, Melk A, et al.: Bbeta(15-42) attenuates the effect of ischemia-reperfusion injury in renal transplantation. J Am Soc Nephrol 22 (10): 1887–1896, 2011.
18. Roesner JP, Petzelbauer P, Koch A, Tran N, Iber T, Vagts DA, Scheeren TW, Vollmar B, Noldge-Schomburg GE, Zacharowski K: Bbeta15-42 (FX06) reduces pulmonary, myocardial, liver, and small intestine damage in a pig model of hemorrhagic shock and reperfusion. Crit Care Med 37 (2): 598–605, 2009.
19. Jennewein C, Mehring M, Tran N, Paulus P, Ockelmann PA, Habeck K, Latsch K, Scheller B, Zacharowski K, Mutlak H: The fibrinopeptide bbeta15–42 reduces inflammation in mice subjected to polymicrobial sepsis. Shock 38 (3): 275–280, 2012.
20. Krishnamoorthy A, Ajay AK, Hoffmann D, Kim TM, Ramirez V, Campanholle G, Bobadilla NA, Waikar SS, Vaidya VS: Fibrinogen beta-derived Bbeta(15–42) peptide protects against kidney ischemia/reperfusion injury. Blood 118 (7): 1934–1942, 2011.
21. Liu A, Dirsch O, Fang H, Sun J, Jin H, Dong W, Dahmen U: HMGB1 in ischemic and non-ischemic liver after selective warm ischemia/reperfusion in rat. Histochem Cell Biol 135 (5): 443–452, 2011.
22. Moloney WC, Mcpherson K, Fliegelman L: Esterase activity in leukocytes demonstrated by the use of naphthol AS-D chloroacetate substrate. J Histochem Cytochem 8: 200–207, 1960.
23. Schlayer HJ, Woort-Menker M, Eyhorn S, Becker H, Schaefer HE, Decker K: Beta-glucuronidase and chloroacetate-esterase staining discriminates rat liver sinusoidal endothelial cells from Kupffer cells in primary culture. Virchows Arch B Cell Pathol Incl Mol Pathol 55 (4): 225–232, 1988.
24. Leder LD: Diagnostic experiences with the naphthol AS-D chloroacetate esterase reaction. Blut 21 (1): 1–8, 1970.
25. Liu A, Dirsch O, Fang H, Dong W, Jin H, Huang H, Sun J, Dahmen U: HMGB1 translocation and expression is caused by warm ischemia reperfusion injury, but not by partial hepatectomy in rats. Exp Mol Pathol 91 (2): 502–508, 2011.
26. Wiedemann D, Schneeberger S, Friedl P, Zacharowski K, Wick N, Boesch F, Margreiter R, Laufer G, Petzelbauer P, Semsroth S: The fibrin-derived peptide Bbeta(15-42) significantly attenuates ischemia-reperfusion injury in a cardiac transplant model. Transplantation 89 (7): 824–829, 2010.
27. Tsung A, Sahai R, Tanaka H, Nakao A, Fink MP, Lotze MT, Yang H, Li J, Tracey KJ, Geller DA, et al.: The nuclear factor HMGB1 mediates hepatic injury after murine liver ischemia-reperfusion. J Exp Med 201 (7): 1135–1143, 2005.
28. Bradham CA, Stachlewitz RF, Gao W, Qian T, Jayadev S, Jenkins G, Hannun Y, Lemasters JJ, Thurman RG, Brenner DA: Reperfusion after liver transplantation in rats differentially activates the mitogen-activated protein kinases. Hepatology 25 (5): 1128–1135, 1997.
29. Drew AF, Liu H, Davidson JM, Daugherty CC, Degen JL: Wound-healing defects in mice lacking fibrinogen. Blood 97 (12): 3691–3698, 2001.
30. Ryu JK, Davalos D, Akassoglou K: Fibrinogen signal transduction in the nervous system. J Thromb Haemost 7 (suppl 7): 151–154, 2009.
31. Harada N, Okajima K, Uchiba M: Dalteparin, a low molecular weight heparin, attenuates inflammatory responses and reduces ischemia-reperfusion–induced liver injury in rats. Crit Care Med 34 (7): 1883–1891, 2006.
32. Loike JD, Sodeik B, Cao L, Leucona S, Weitz JI, Detmers PA, Wright SD, Silverstein SC: CD11c/CD18 on neutrophils recognizes a domain at the N terminus of the A alpha chain of fibrinogen. Proc Natl Acad Sci U S A 88 (3): 1044–1048, 1991.
33. Bustin M, Reeves R: High-mobility-group chromosomal proteins: architectural components that facilitate chromatin function. Prog Nucleic Acid Res Mol Biol 54: 3–100 5–100, 1996.
34. Bianchi ME, Beltrame M, Paonessa G: Specific recognition of cruciform DNA by nuclear protein HMG1. Science 243 (4894 Pt 1): 1056–1059, 1989.
35. Klune JR, Dhupar R, Cardinal J, Billiar TR, Tsung A: HMGB1: endogenous danger signaling. Mol Med 14 (7–8): 476–484, 2008.
36. Lotze MT, Tracey KJ: High-mobility group box 1 protein (HMGB1): nuclear weapon in the immune arsenal. Nat Rev Immunol 5 (4): 331–342, 2005.
37. Czaja MJ, Liu H, Wang Y: Oxidant-induced hepatocyte injury from menadione is regulated by ERK and AP-1 signaling. Hepatology 37 (6): 1405–1413, 2003.
38. Uehara T, Bennett B, Sakata ST, Satoh Y, Bilter GK, Westwick JK, Brenner DA: JNK mediates hepatic ischemia reperfusion injury. J Hepatol 42 (6): 850–859, 2005.
39. Kobayashi M, Takeyoshi I, Yoshinari D, Matsumoto K, Morishita Y: P38 mitogen-activated protein kinase inhibition attenuates ischemia-reperfusion injury of the rat liver. Surgery 131 (3): 344–349, 2002.
Keywords:

Liver ischemia/reperfusion; fibrinogen; Bβ15-42; inflammatory response; HMGB1; MAPK

©2013The Shock Society