J Korean Med Sci.  2009 Oct;24(5):775-781. 10.3346/jkms.2009.24.5.775.

The Role of Keratinocyte-derived Chemokine in Hemorrhage-induced Acute Lung Injury in Mice

• Lee BH
• Lee TJ
• Jung JW
• Oh DJ
• Choi JC
• Shin JW
• Park IW
• Choi BW
• Kim JY

• KMID: 1781993
• DOI: http://doi.org/10.3346/jkms.2009.24.5.775

Abstract

Dominant inflammatory cytokines might be different depending on the underlying causes of acute lung injury (ALI). The role of kertinocyte-derived chemokine (KC), a potent chemoattractant for neutrophils, has not been clearly established in hemorrhage-induced ALI. In this study, lung injury and cytokine expressison were evaluated in LPS- or hemorrhage-induced ALI models of BALB/c mice. The myeloperoxidase activities at 4 hr after hemorrhage and LPS-injection were 47.4+/-13.0 and 56.5+/-16.4 U/g, respectively. NF-kappa B activity peaked at 4 hr after hemorrhage, which was suppressed to the control level by anti-high mobility group B1 (HMGB1) antibody. Lung expressions of TNF-alpha, MIP-2, and IL-1beta were increased by LPS injection. However, there was only a minimal increase in IL-1beta and no expressions of TNF-alpha or MIP-2 in hemorrhage-induced ALI. In contrast, lung KC increased significantly at 4 hr after hemorrhage compared to control levels (83.1+/-12.3 vs. 14.2+/-1.6 pg/mL/mg by ELISA) (P<0.05). By immunohistochemical staining, lung neutrophils stained positive for KC. Increased KC was also observed in bronchoalveolar lavage fluid and plasma. KC plays an important role in hemorrhage-induced ALI.

Keyword

Acute Lung Injury; LPS; Hemorrhage; Keratinocyte-derived Chemokines

MeSH Terms

Acute Lung Injury/etiology/*metabolism
Animals
Antibodies/immunology/metabolism
Chemokine CXCL2/analysis
Chemokines/analysis/blood/*physiology
Chickens
HMGB1 Protein/metabolism
Humans
Interleukin-1beta/analysis
Lipopolysaccharides/toxicity
Mice
Mice, Inbred BALB C
NF-kappa B/metabolism
Neutrophils/immunology/metabolism
Peroxidase/analysis
Shock, Hemorrhagic/*complications
Time Factors
Tumor Necrosis Factor-alpha/analysis

Figure

• 

  Fig. 1 Lung neutrophil accumulation as assessed by MPO activity after hemorrhage- or endotoxemia-induced acute lung injury. Neutrophil accumulation in the lung was evaluated with MPO assay at 0, 4 hr, 24 hr, 48 hr and 72 hr after hemorrhage and 4 hr after lipopolysaccharide (LPS) injection (5 mg/kg, i.p.). Data are mean±SEM of at least 5 mice per group. *P<0.05.

• 

  Fig. 2 Anti-HMGB1 antibody treatment decreases the NF-κB activation in the lung. One hour after the induction of hemorrhage, mice were injected with either control chicken IgY antibody (Hem+Cont Ab, 200 µg/mice) or anti-HMGB1 antibody (Hem+anti-HMGB1, 200 µg/mice). Four hours after the induction of hemorrhage, mice were sacrificed and NF-κB activity was determined in the lung with EMSA. The data shown are representative of three independent experiments.

• 

  Fig. 3 LPS induces an increase in the expression of TNF-α, MIP-2 and IL-1β in the lung. The expressions of TNF-α, MIP-2 and IL-1β of lung tissue were determined by ELISA at 4 hr after injection of LPS (5 mg/kg, i.p.). Data are mean±SEM of at least 5 mice per group. *P<0.05.

• 

  Fig. 4 Hemorrhage induces minimal increase of IL-1β and no increase of TNF-α and MIP-2 expression in the lung. The expressions of TNF-α, MIP-2 and IL-1β of lung tissue were determined by ELISA at 0, 4 hr, 24 hr, 48 hr, and 72 hr after hemorrhage. Data are mean±SEM of at least 5 mice per group. *P<0.05.

• 

  Fig. 5 Hemorrhage induces an increase in the expression of KC in the lung. The expression of KC of lung tissue was determined by ELISA at 0, 4 hr, 24 hr, 48 hr, and 72 hr after hemorrhage. Data are mean±SEM of at least 5 mice per group. *P<0.05.

• 

  Fig. 6 Increased expression of KC in the lung at 4 hr after hemorrhage was confirmed by immunohistochemical staining. The expression of KC of lung tissue was determined by immunohistochemical (IHC) staining at 4 hr after hemorrhage. Rabbit polyclonal GRO alpha (1:50, Abcam, Cambridge, U.K.) was used as a primary antibody. Biotinated, anti-rabbit antibody (1:300, Zymed, South San Francisco, CA, U.S.A.) was applied as a secondary antibody. For background staining, 3-amino-9-ethyl carbazole (AEC) was used (×400).

• 

  Fig. 7 Hemorrhage induces an increase in the expression of KC in bronchoalveolar lavage (BAL) fluid. The expression of KC of BAL fluid was determined by ELISA at 4 hr and 24 hr after hemorrhage. Data are mean±SEM of at least 5 mice per group. *P<0.05.

• 

  Fig. 8 Hemorrhage induces an increase in plasma expression of KC. The expressions of KC of plasma was determined by ELISA at 0, 4 hr, 24 hr, 48 hr, and 72 hr after hemorrhage. Data are mean±SEM of at least 5 mice per group. *P<0.05.

Reference

1. Bernard GR, Artigas A, Brigham KL, Carlet J, Falke K, Hudson L, Lamy M, Legall JR, Morris A, Spragg R. The American-European consensus conference on ARDS. Definition, mechanisms, relevant outcomes, and clinical trial coordination. Am J Respir Crit Care Med. 1994. 149:818–824.

2. Ashbaugh DG, Bigelow DB, Petty TL, Levine BE. Acute respiratory distress in adults. Lancet. 1967. 2:319–323.

3. Rubenfeld GD, Caldwell E, Peabody E, Weaver J, Martin DP, Neff M, Stern EJ, Hudson LD. Incidence and outcomes of acute lung injury. N Engl J Med. 2005. 353:1685–1693.

4. Ware LB, Matthay MA. The acute respiratory distress syndrome. N Engl J Med. 2000. 342:1334–1349.

5. Sevransky JE, Levy MM, Marini JJ. Mechanical ventilation in sepsis-induced acute lung injury/acute respiratory distress syndrome: an evidence-based review. Crit Care Med. 2004. 32(11):Suppl. S548–S553.

6. Hudson LD, Milberg JA, Anardi D, Maunder RJ. Clinical risks for development of the acute respiratory distress syndrome. Am J Respir Crit Care Med. 1995. 151:293–301.

7. Abraham E, Carmody A, Shenkar R, Arcaroli J. Neutrophils as early immunologic effectors in hemorrhage- or endotoxemia-induced acute lung injury. Am J Physiol Lung Cell Mol Physiol. 2000. 279:L1137–L1145.

8. Molls RR, Savransky V, Liu M, Bevans S, Mehta T, Tuder RM, King LS, Rabb H. Keratinocyte-derived chemokine is an early biomarker of ischemic acute kidney injury. Am J Physiol Renal Physiol. 2006. 290:1187–1193.

9. Schober A. Chemokines in vascular dysfunction and remodeling. Arterioscler Thromb Vasc Biol. 2008. 28:1950–1959.

10. Huang S, Paulauskis JD, Godleski JJ, Kobzik L. Expression of macrophage inflammatory protein-2 and KC mRNA in pulmonary inflammation. Am J Pathol. 1992. 141:981–988.

11. Frink M, Hsieh YC, Hsieh CH, Pape HC, Choudhry MA, Schwacha MG, Chaudry IH. Keratinocyte-derived chemokine plays a critical role in the induction of systemic inflammation and tissue damage after trauma-hemorrhage. Shock. 2007. 28:576–581.

12. Kim JY, Park JS, Strassheim D, Douglas I, Diaz del Valle F, Asehnoune K, Mitra S, Kwak SH, Yamada S, Maruyama I, Ishizaka A, Abraham E. HMGB1 contributes to the development of acute lung injury after hemorrhage. Am J Physiol Lung Cell Mol Physiol. 2005. 288:L958–L965.

13. Choi JC, Jung JW, Kwak HW, Song JH, Jeon EJ, Shin JW, Park IW, Choi BW, Kim JY. Granulocyte macrophage-colony stimulating factor (GM-CSF) augments acute lung injury via its neutrophil priming effects. J Korean Med Sci. 2008. 23:288–295.

14. Schell MT, Spitzer AL, Johnson JA, Lee D, Harris HW. Heat shock inhibits NF-kB activation in a dose- and time-dependent manner. J Surg Res. 2005. 129:90–93.

15. Barsness KA, Arcaroli J, Harken AH, Abraham E, Banerjee A, Reznikov L, McIntyre RC. Hemorrhage-induced acute lung injury is TLR-4 dependent. Am J Physiol Regul Integr Comp Physiol. 2004. 287:R592–R599.

16. Brundage SI, Schreiber MA, Holcomb JB, Zautke N, Mastrangelo MA, Xq X, Macaitis J, Tweardy DJ. Amplification of the proinflammatory transcription factor cascade increases with severity of uncontrolled hemorrhage in swine. J Surg Res. 2003. 113:74–80.

17. Andersson U, Erlandsson-Harris H, Yang H, Tracey KJ. HMGB1 as a DNA-binding cytokine. J Leukoc Biol. 2002. 72:1084–1091.

18. Yang H, Wang H, Czura CJ, Tracey KJ. The cytokine activity of HMGB1. J Leukoc Biol. 2005. 78:1–8.

19. Abraham E, Arcaroli J, Carmody A, Wang H, Tracey KJ. HMG-1 as a mediator of acute lung inflammation. J Immunol. 2000. 165:2950–2954.

20. Ombrellino M, Wang H, Ajemian MS, Talhouk A, Scher IA, Friedman SG, Tracey KJ. Increased serum concentration of high-mobility-group protein 1 in hemorrhagic shock. Lancet. 1999. 354:1446–1447.

21. Lomas-Neira JL, Chung CS, Grutkoski PS, Miller EJ, Ayala A. CXCR2 inhibition suppresses hemorrhage-induced priming for acute lung injury in mice. J Leukoc Biol. 2004. 76:58–64.