help button home button
AJRCCM
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow A correction has been published
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by BOWLER, R. P.
Right arrow Articles by ABRAHAM, E.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by BOWLER, R. P.
Right arrow Articles by ABRAHAM, E.
Am. J. Respir. Crit. Care Med., Volume 164, Number 2, July 2001, 290-294

Extracellular Superoxide Dismutase Attenuates Lung Injury after Hemorrhage

RUSSELL P. BOWLER, JOHN ARCAROLI, JAMES D. CRAPO, ARON ROSS, JAN W. SLOT, and EDWARD ABRAHAM

National Jewish Medical and Research Center, Denver, Colorado; Division of Pulmonary Sciences and Critical Care Medicine, University of Colorado Health Sciences Center, Denver, Colorado; and Department of Pathology, University of Utrecht, Utrecht, The Netherlands




    ABSTRACT
TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES

Reperfusion of the lung after hemorrhage generates free radicals such as superoxide (O2·) that may injure the lung; however, the relative importance of intracellular versus extracellular free radicals is unclear. The superoxide dismutases (SOD) are the primary enzymatic method to reduce superoxide. We examined whether lung-specific overexpression of extracellular superoxide dismutase (EC-SOD) would attenuate hemorrhage-induced lung injury. Wild-type mice and mice overexpressing the human EC-SOD gene with a lung-specific promoter were hemorrhaged by removing 30% of blood volume. After hemorrhage, the lung wet to dry weight ratios increased from 5.4 ± 0.11 in unmanipulated control mice to 6.3 ± 0.16 in wild-type mice, but to only 5.60 ± 0.17 in the EC-SOD transgenic mice (p < 0.05 compared with hemorrhaged wild-type). Hemorrhage-induced lipid peroxidation, as assessed by lung F2 isoprostanes, was lower in the EC-SOD transgenic mice (3.4 ± 0.3 µg/lung) compared with wild-type mice (1.9 ± 0.2 µg/lung; p < 0.05). Compared with wild-type, EC-SOD transgenic mice had attenuated the hemorrhage-induced increase in both pulmonary nuclear factor kappa B (NK-kappa B) activation (relative absorbance 1.1 ± 0.2 for EC-SOD transgenic versus 2.5 ± 0.1 for wild-type; p < 0.05) and myeloperoxidase activity (5.1 ± 0.87 units/g for EC-SOD transgenic versus 11.3 ± 1.8 units/g for wild-type; p < 0.01). Thus, overexpression of pulmonary EC-SOD in the mouse lung attenuates lung injury after hemorrhage.

Keywords: hemorrhage; extracellular superoxide dismutase; NF-kappa B; myeloperoxidase



    INTRODUCTION
TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES

Acute lung injury (ALI) frequently occurs after blood loss (1). Reactive oxygen species (ROS) generated as a result of ischemia-reperfusion associated with hemorrhage have been postulated to contribute to the development of ALI in this setting (2). Both an activation of membrane oxidases (3) and the uncoupling of the respiratory chain in mitochondria (4) contribute to ROS production after reperfusion. Superoxide is one of the ROS that mediate this injury.

The superoxide dismutase (SOD) family catalyzes the dismutation of superoxide into hydrogen peroxide and oxygen at an extremely rapid rate (k > 109 M-1s-1) (5). Three SOD enzymes (intracellular [CuZnSOD or SOD1], mitochondrial [MnSOD or SOD2], and extracellular [EC-SOD or SOD3]) have been identified in mammals. Superoxide is likely to be especially important in the extracellular space of the lung because lung tissue has one of the highest concentrations of EC-SOD (5). The distribution of EC-SOD in normal lung suggests that control of superoxide is important in specific extracellular compartments. In normal lung, EC-SOD immunolocalizes to the alveolar interstitium and extracellular space, but not pulmonary endothelial cells (6). Furthermore, EC-SOD is unique among SOD enzymes in that it has a carboxyterminus with high affinity for the extracellular matrix (ECM). Thus, extracellular superoxide would seem to be particularly important in controlling superoxide in the alveolar interstitium.

Although others have investigated the role of superoxide in hemorrhage, their focus has been on vascular or intracellular compartments. For instance, hemorrhage-induced superoxide has recently been found to play a role in both the intracellular space and on the endothelial surface (7), and exogenously administered SOD, which sticks to endothelium, can ameliorate a hemorrhage-induced decrease in blood pressure (8) and survival (9). However, the role of ROS in the ECM of the alveolar interstitium, particularly for superoxide, has not been fully elucidated.

Our laboratory has constructed an EC-SOD transgenic mouse that specifically overexpresses EC-SOD in the lung in order to study the relationship between superoxide, the extracellular space, and lung injury. In the present investigation, we examined the ability of EC-SOD to attenuate hemorrhage- induced lung injury during oxidative stress. Using a mouse that overexpressed human EC-SOD under the control of the surfactant protein C (SPC) promoter, the effect of selectively increasing EC-SOD in the lung on the development of ALI was evaluated.


    METHODS
TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES

Experiments were conducted in accordance with institutional review board-approved protocols. Sixteen mice overexpressing full-length human EC-SOD complementary DNA (cDNA) using a SPC promoter that directs transgene expression to alveolar type II and nonciliated distal bronchial epithelial cells (12) and 33 negative (wild-type) littermates 8 to 12 wk of age were included in the hemorrhaged and untreated control groups.

Hemorrhage was performed as previously described (13). Thirty percent of the blood volume calculated by weight (0.55 ml per 20 g mouse) was removed by cardiac puncture. Mice were killed after 1 h.

EC-SOD Quantitation

A lobe of the lung was homogenized in lysis buffer (Tris-Cl 25 mM, pH 7.5; NaCl 500; ethylenediaminetetraacetic acid [EDTA] 5 mM; triton 0.5%). A volume of 10 µg was used for a Western blot (rabbit polyclonal alpha -EC-SOD antibody) that was quantitated using ECL Plus (Amersham Pharmacia Biotech, Piscataway, NJ) and a Molecular Dynamics Imaging system (Amersham Pharmacia Biotech).

Immunocytochemistry

Multiple EC-SOD transgenic mouse lungs were inflation fixed in a mixture of 2% formaldehyde and 0.2% glutaraldehyde in 0.05 M phosphate-buffered saline (PBS) for 1 h followed by overnight fixation in 4% paraformaldehyde at 4° C (14). After fixation, the tissues were equilibrated in 30% sucrose overnight at 4° C. Ultrathin cryosections of EC-SOD transgenic mouse lung tissue were immunolabeled with rabbit antirecombinant human-EC-SOD and 10 nm protein A-gold (6). After immunolabeling, the sections were stained and embedded in mixtures of uranyl acetate and methylcellulose (15).

Wet-to-Dry Lung Weight Ratios

Lungs were excised, rinsed briefly in PBS, blotted, then weighed to obtain the "wet" weight. Lungs were dried in an oven at 80° C for 7 d to obtain the "dry" weight.

Myeloperoxidase (MPO)

Using a modification from Anderson and coworkers (16, 17), a lobe of the lung from each animal was homogenized for 30 s in 1.5 ml 20 mM potassium phosphate, pH 7.4, and centrifuged at 4° C for 30 min at 40,000 × g. The pellet was resuspended in 1.5 ml 40 mM potassium phosphate, pH 6.0, containing 0.5% hexacetyltrimethyl ammonium bromide, sonicated for 90 s, incubated at 60° C for 2 h, and centrifuged. The supernatant was assayed for peroxidase activity corrected to lung weight.

Electrophoretic Mobility Shift Assay (EMSA) Analysis of Nuclear Factor Kappa B (NK-kappa B)

Nuclear extracts were prepared as previously described (18, 19). Activation of NF-kappa B was determined by EMSA analysis (2, 20, 21).

Determination of Lipid Peroxidation

The lipid fraction was extracted from homogenized tissue, partially purified by solid-phase extraction and then derivatized to the pentafluorobenzyl ester trimethylsilyl ethers. F2-isoprostanes were measured by negative ion chemical ionization gas chromatography/mass spectrometry (GC/MS) analysis (22).

Statistical Analysis

A Tukey-Kramer multiple group comparison test was used to compare individual groups. All values were calculated using GraphPad Prism version 3.00 for Windows (GraphPad Software, San Diego, CA).


    RESULTS
TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES

Characterization of the EC-SOD Transgenic Mice

Compared with wild-type mice, mice overexpressing the EC-SOD transgene have a threefold increase in lung tissue EC-SOD (Figure 1), yet have been previously shown to have no difference in SOD1, SOD2, glutathione peroxidase, glutathione, and catalase (12).



View larger version (22K):
[in this window]
[in a new window]
  		Figure 1.   EC-SOD protein expression in wild-type and transgenic mouse lungs. Western blots reveal that transgene mice have a threefold increase in lung EC-SOD protein. The antibody recognizes both mouse EC-SOD (mEC-SOD) and human EC-SOD (hEC-SOD) protein. The molecular weight of hEC-SOD is less than mEC-SOD.

The distribution of EC-SOD within the alveolar septum was evaluated from multiple EC-SOD transgenic mouse lungs using immunocytochemistry (Figure 2). The antibody was specific for human EC-SOD and did not label wild-type lungs. This micrograph clearly demonstrates that the transgene product is mainly associated with ECM elements in the interstitial space of the alveoli. There was little intracellular labeling. The luminal endothelial cell surface, which is rich in heparin proteoglycans, did not have significant EC-SOD labeling. The procedure did not preserve airspace surfactant, so we were not able to assess the level of EC-SOD transgene product on the surface of alveolar epithelial cells. Thus, the transgenic mice have a threefold increase in EC-SOD that immunolocalizes primarily in the alveolar interstitium and is associated with the ECM.



View larger version (153K):
[in this window]
[in a new window]
  		Figure 2.   Electron microscopy with gold labeling in a mouse overexpressing human EC-SOD indicates that EC-SOD (small arrows) can be found in the alveolar interstitial space (I ) between endothelial (En) and epithelial cells (Ep). EC-SOD is also associated with ECM elements (large arrows). EC-SOD is not associated with red blood cells (RBC), white blood cells (WBC), or airspace (A) (Bar 200 nm).

Reduction of Lung Injury with Lung EC-SOD Overexpression

Lung wet-to-dry weight ratios were 5.5 ± 0.11 in unmanipulated mice and increased to 6.3 ± 0.16 after hemorrhage (Figure 3). However, in EC-SOD transgenic mice, the wet-dry ratios were 5.60 ± 0.17 after hemorrhage (p < 0.05 comparing wild-type to EC-SOD transgenic).



View larger version (11K):
[in this window]
[in a new window]
  		Figure 3.   Lung edema. Lung wet-to-dry ratios were measured in either unmanipulated mice (control) or wild-type mice and EC-SOD transgenic mice 1 h after 30% of blood volume had been removed. Hemorrhage significantly increased the wet-to-dry ratio in the wild-type mouse lung, but this increase was significantly attenuated in mice overexpressing EC-SOD in the lungs. * p < 0.05. Bar indicates SEM.

Hemorrhage-induced neutrophil accumulation in the lungs was also reduced in EC-SOD transgenic mice compared with wild-type mice (Figure 4). Lung MPO was 1.1 ± 0.1 units/g in unmanipulated mouse lungs, 11.3 ± 1.8 units/g after hemorrhage in the wild-type lungs, and 5.1 ± 0.87 units/g after hemorrhage in the EC-SOD transgenic mouse lungs (p < 0.01 comparing hemorrhaged wild-type with EC-SOD transgenic; p < 0.001 comparing unmanipulated mice with hemorrhaged wild-type).



View larger version (9K):
[in this window]
[in a new window]
  		Figure 4.   Lung MPO activity was measured in either unmanipulated mice (control) or wild-type mice and EC-SOD transgenic mice 1 h after 30% of blood volume had been removed. Hemorrhage significantly increased the MPO activity in the wild-type mouse lung, but this increase was significantly attenuated in mice overexpressing EC-SOD in the lungs. **p < 0.01; ***p < 0.001. Bar indicates SEM.

Overexpression of EC-SOD Decreases Lipid Peroxidation

Hemorrhage has previously been shown to increase lipid peroxidation in the lungs (23). F2-isoprostanes are free radical catalyzed prostaglandin isomers that increase in tissue during oxidative stress (24). To determine the role of EC-SOD in modulating such oxidant-induced lung injury, we measured lung F2-isoprostanes in EC-SOD transgenic and wild-type mice before and after blood loss (Figure 5).



View larger version (12K):
[in this window]
[in a new window]
  		Figure 5.   Lung F2-isoprostanes were measured in either unmanipulated mice (control) or wild-type mice and EC-SOD transgenic mice 1 h after 30% of blood volume had been removed. Hemorrhage significantly increased F2-isoprostanes in the wild-type mouse lung, but this increase was significantly attenuated in mice overexpressing EC-SOD in the lungs. *p < 0.05. Bar indicates SEM.

F2-isoprostanes in lung increased from 2.3 ± 0.2 µg/lung in unmanipulated mice to 3.4 ± 0.3 µg/lung after hemorrhage in the wild-type (p < 0.05 compared with unmanipulated mice). In the EC-SOD transgene positive mice, there was a smaller increase in isoprostanes after hemorrhage (2.0 ± 0.2 µg/lung; p < 0.05 comparing hemorrhaged wild-type with EC-SOD transgenic; p < 0.05 comparing unmanipulated mice with hemorrhaged wild-type).

Overexpression of EC-SOD Decreases NF-kappa B Activation

Hemorrhage results in activation of the transcriptional regulatory factor NF-kappa B in the lungs (2) through pathways involving ROS (25, 26). To examine the role of EC-SOD in modulating hemorrhage-induced NF-kappa B activation, we measured lung NF-kappa B activity in wild-type and EC-SOD transgenic mice after blood loss (Figure 6). After quantitation of NF-kappa B activity using densitometry, values were normalized to unmanipulated wild-type mice. Overexpression of EC-SOD attenuated hemorrhage-induced NF-kappa B activation compared with wild-type mouse lungs (relative absorbance 1.3 ± 0.1 versus 2.5 ± 0.3; p < 0.05).



View larger version (18K):
[in this window]
[in a new window]
  		Figure 6.   Pulmonary NF-kappa B activation 1 h after hemorrhage. (A) Scanning densitometry of NF-kappa B analysis is shown for each group with the average density of the unhemorrhaged (control) set equal to one. (B) A representative experiment is shown. Two additional experiments with separate sets of animals gave similar results. *p < 0.05; **p < 0.01. Bar indicates SEM.

EC-SOD Protein in Hemorrhaged Mice

Proinflammatory cytokines, including interleukin-1beta , tumor necrosis factor-alpha , and macrophage inflammatory protein-2 are increased in the lungs after hemorrhage (2, 13, 17). The cytokines interleukin-1beta , tumor necrosis factor-alpha , and interferon-gamma increase EC-SOD protein in cell culture (27, 28). As shown in Figure 1, EC-SOD transgenic mice have a threefold increase in lung tissue EC-SOD protein. To see if hemorrhage induced a potentiating effect on EC-SOD expression in vivo, we examined lung EC-SOD protein after blood loss. Hemorrhage did not significantly increase total lung EC-SOD protein in either the wild-type (6 ± 10% increase) or EC-SOD transgenic mice (15 ± 25% increase).


    DISCUSSION
TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES

Hemorrhage causes whole body ischemia with resultant generation of ROS when reperfusion occurs (29). In the lung, ROS are capable not only of potentiating neutrophil recruitment (30), but also are associated with injury to proteins, lipids, and DNA (24, 31). Superoxide production is increased after blood loss and contributes to oxidant-induced tissue injury in this setting (8, 34, 35). SODs are the primary defense against excess superoxide.

The consequences of the superoxide/SOD interaction are likely to be highly dependent on the domain in which they occur because superoxide is impermeable to membranes and the three mammalian SODs are fixed within their subcellular localization. The specific lung domains likely to be relevant to reperfusion injury include: the endothelial membrane and the vascular space; the cytoplasm of endothelial cells, smooth muscle cells, or alveolar epithelial cells; mitochondria; the alveolar interstitium and its ECM; and the alveolar epithelial membrane and the alveolar airspace. Multiple published reports address the role of the first two domains, but we are unaware of any reports specifically examining the role of superoxide in the alveolar interstitium and ECM after hemorrhage.

The role of superoxide in activating the endothelial membrane and initiating neutrophil adhesion is well established (36). For instance, exogenous SOD1 blunts the vascular expression of P-selectin, an adhesion molecule crucial for recruitment of activated leukocytes to the lung (7). Other adhesion molecules, such as E-selectin, can also be inhibited in vitro by adding SOD1 (37). These pharmacologic experiments used SOD1, an enzyme that is normally found in the cytoplasm. However, in these experiments it is likely that most of the added SOD activity remained extracellularly. Additionally, SOD1 has a net negative charge, unlike EC-SOD, whose carboxyterminus gives it a strong positive charge that determines distribution. Finally, because there is normally scant SOD found on the surface of pulmonary vessels (6), it is unclear how much SOD modulates hemorrhage-induced superoxide production in vivo. Thus, the distribution of EC-SOD activity in our EC-SOD transgenic animals is likely very different from that in the pharmacologic experiments. The cytoplasm, on the other hand, has high levels of superoxide activity. Overexpression of cytoplasmic SOD has been shown to blunt activation of endothelium after hemorrhage (7), but these experiments did not elucidate in which cells overexpression was important.

This is the first report to specifically address the role of superoxide in the alveolar interstitium and ECM. We found that overexpression of EC-SOD could attenuate several markers of lung injury after hemorrhage: lung edema, lipid peroxidation, and MPO activity. Lung edema has been previously shown to increase after lung injury and likely represents increased microvascular permeability (24, 38). Lipid peroxidation, as assessed by F2-isoprostanes, has also been demonstrated to increase after hemorrhage. For example, blood loss due to aortic rupture produces elevated plasma F2-isoprostanes in humans (29) and increased F2-isoprostanes in the lungs of rats (23). MPO causes much of the lung damage associated with hemorrhage (38). Thus, a reduction of all of these markers in the EC-SOD transgenic mouse implies that superoxide activity in the alveolar interstitium and ECM is an important mediator in hemorrhage-induced lung injury.

EC-SOD probably reduces lung injury by decreasing superoxide in the alveolar interstitium and ECM. This may lead to decreased recruitment of activated neutrophils to the lung by an unclear mechanism. Our data suggest that it may be mediated by reducing activity of NF-kappa B. We have recently reported that neutrophils are responsible for most of the increased activity of NF-kappa B in the lungs after hemorrhage (41). Thus, the attenuated increase in lung NF-kappa B activity in the EC-SOD transgenic mouse lung is most likely to a decrease in activated lung neutrophils. Whether neutrophils have a direct or indirect interaction with alveolar interstitial superoxide is currently the subject of investigation in our laboratory.

The link between activation of inflammatory cells and EC-SOD is strengthened by the observation that NF-kappa B controls transcriptional regulation of EC-SOD in alveolar type II cells (27). Because hemorrhage results in rapid activation of NF-kappa B in the lungs, we expected to find increased EC-SOD in this setting. However, no hemorrhage-induced alteration in lung EC-SOD was found in the present experiments. It is possible that the time elapsed posthemorrhage was too short to permit increased expression of EC-SOD. In vitro, maximal transcription of the EC-SOD gene only occurs 6 h after NF-kappa B activation is induced (27) and optimal EC-SOD protein secretion only occurs after 3 d of continuous cytokine stimulation (28).

This study has potentially important implications in the prevention and treatment of acute respiratory distress syndrome after hemorrhagic shock. Although previous studies have demonstrated the importance of superoxide in reperfusion injury (7, 34), this is the first study to show that EC-SOD can attenuate hemorrhage-induced lung injury and suggests that the alveolar interstitial space and ECM play a role in superoxide-mediated lung injury. Our data suggest that elevated alveolar interstitial SOD activity may decrease the likelihood of developing acute inflammatory lung injury after hemorrhage. However, because the present experiments used a model where increased lung concentrations of EC-SOD were present before hemorrhage, the potential clinical benefit of EC-SOD administration after blood loss remains to be proven.


    Footnotes

Correspondence and requests for reprints should be addressed to Russell P. Bowler, M.D., National Jewish Medical and Research Center, K707, 1400 Jackson Street, Denver, CO 80206. E-mail: BowlerR{at}njc.org

(Received in original form November 13, 2000 and in revised form March 30, 2001).

Acknowledgments: Supported by Grants NIH HL-04407, HL-62221, HL31992, HL 42444, and the Andrew Goodman Fellowship in Medicine at National Jewish Medical and Research Center.
    References
TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES

1. Rainer TH, Lam PK, Wong EM, Cocks RA. Derivation of a prediction rule for post-traumatic acute lung injury. Resuscitation 1999; 42: 187-196 [Medline].

2. Shenkar R, Abraham E. Mechanisms of lung neutrophil activation after hemorrhage or endotoxemia: roles of reactive oxygen intermediates, NF-kappa B, and cyclic AMP response element binding protein. J Immunol 1999; 163: 954-962 [Abstract/Free Full Text].

3. Henderson LM, Chappel JB. NADPH oxidase of neutrophils. Biochim Biophys Acta 1996; 1273: 87-107 [Medline].

4. Ambrosio G, Zweier JL, Duilio C, Kuppusamy P, Santoro G, Elia PP, Tritto I, Cirillo P, Condoreli M, Chiariello M, et al . Evidence that mitochondrial respiration is a source of potentially toxic oxygen free radicals in intact rabbit hearts subjected to ischemia and reflow. J Biol Chem 1993; 268: 18532-18541 [Abstract/Free Full Text].

5. Marklund SL. Human copper-containing superoxide dismutase of high molecular weight. Proc Natl Acad Sci USA 1982; 79: 7634-7638 [Abstract/Free Full Text].

6. Oury TD, Chang LY, Marklund SL, Day BJ, Crapo JD. Immunocytochemical localization of extracellular superoxide dismutase in human lung. Lab Invest 1994; 70: 889-898 [Medline].

7. Akgur FM, Brown MF, Zibari GB, McDonald JC, Epstein CJ, Ross CR, Granger DN. Role of superoxide in hemorrhagic shock-induced P-selectin expression. Am J Physiol Heart Circ Physiol 2000; 279: H791-797 [Abstract/Free Full Text].

8. Simon HM, Scalea T, Paskanik A, Yang B. Superoxide dismutase (SOD) prevents hypotension after hemorrhagic shock and aortic cross clamping. Am J Med Sci 1996; 312: 155-159 [Medline].

9. Rhee P, Waxman K, Clark L, Tominaga G, Soliman MH. Superoxide dismutase poilyethylene glycol improves survival in hemorrhagic shock. Am Surg 1991; 57: 747-750 [Medline].

10. Tan LR, Waxman K, Clark L, Eloi L, Chhieng N, Miller B, Young A. Superoxide dismutase and allopurinol improve survival in an animal model of hemorrhagic shock. Am Surg 1993; 59: 797-800 [Medline].

11. Tominaga GT, Bailey S, Daughters K, Sarfeh IJ, Waxman K. The effect of polyethylene glycol-superoxide dismutase on gastric mucosa and survival in shock with tissue injury. Am Surg 61:925-929.

12. Folz RJ, Abushamaa AM, Suliman HB. Extracellular superoxide dismutase in the airways of transgenic mice reduces inflammation and attenuates lung toxicity following hyperoxia. J Clin Invest 103:1055-1066.

13. Abraham E, Freitas AA. Hemorrhage produces abnormalities in lymphocyte function and lymphokine generation. J Immunol 142:899-906.

14. Liou W, Geuze HJ, Slot JW. Improving structural integrity of cryosections for immunogold labeling. Histochem Cell Biol 106:41-58.

15. Crapo JD, Oury T, Rabouille C, Slot JW, Chang LY. Copper, zinc superoxide dismutase is primarily a cytosolic protein in human cells. Proc Natl Acad Sci USA 89:10405-10409.

16. Anderson BO, Brown JM, Shanley PF, Bensard DD, Harken AH. Marginating neutrophils are reversibly adherent to normal lung endothelium. Surgery 109:51-61.

17. Parsey MV, Tuder RM, Abraham E. Neutrophils are major contributors to intraparenchymal lung IL-1 beta expression after hemorrhage and endotoxemia. J Immunol 160:1007-1013.

18. Kelley DM, Lichtenstein A, Wang J, Taylor AN, Dubinett SM. Corticotropin-releasing factor reduces lipopolysaccharide-induced pulmonary vascular leak. Immunopharmacol Immunotoxicol 1994; 16: 139-148 [Medline].

19. Kollef MH, Schuster DP. The acute respiratory distress syndrome. N Engl J Med 1995; 332: 27-37 [Free Full Text].

20. Moine P, McIntyre R, Schwartz MD, Kaneko D, Shenkar R, Le Tulzo Y, Moore EE, Abraham E. NF-kappaB regulatory mechanisms in alveolar macrophages from patients with acute respiratory distress syndrome. Shock 2000; 13: 85-91 [Medline].

21. Shenkar R, Abraham E. Hemorrhage induces rapid in vivo activation of CREB and NF-kappaB in murine intraparenchymal lung mononuclear cells. Am J Respir Cell Mol Biol 1997; 16: 145-152 [Abstract].

22. Waugh RJ, Morrow JD, Roberts LJ 2nd,, Murphy RC. Identification and relative quantitation of F2-isoprostane regioisomers formed in vivo in the rat. Free Radic Biol Med 1997; 23: 943-954 [Medline].

23. Boyd AJ, Rubin BB, Walker PM, Romaschin A, Issekutz TB, Lindsay TF. A CD18 monoclonal antibody reduces multiple organ injury in a model of ruptured abdominal aortic aneurysm. Am J Physiol 1999; 277(1 Pt 2):H172-182.

24. Becker PM, Sanders SP, Price P, Christman BW. F2-isoprostane generation in isolated ferret lungs after oxidant injury or ventilated ischemia. Free Radic Biol Med 1998; 25: 703-711 [Medline].

25. Ogata N, Yamamoto H, Kugiyama K, Yasue H, Miyamoto E. Involvement of protein kinase C in superoxide anion-induced activation of nuclear factor-kappa B in human endothelial cells. Cardiovasc Res 2000; 45: 513-521 [Abstract/Free Full Text].

26. Jaspers I, Samet JM, Erzurum S, Reed W. Vanadium-induced kappaB-dependent transcription depends upon peroxide-induced activation of the p35 mitogen-activated protein kinase. Am J Respir Cell Mol Biol 2000; 23: 95-102 [Abstract/Free Full Text].

27. Brady TC, Chang LY, Day BJ, Crapo JD. Extracellular superoxide dismutase is upregulated with inducible nitric oxide synthase after NF-kappa B activation. Am J Physiol 1997;273(5 PT 1):L1002-1006.

28. Marklund SL. Regulation by cytokines of extracellular superoxide dismutase and other superoxide dismutase isoenzymes in fibroblasts. J Biol Chem 1992; 267: 6696-6701 [Abstract/Free Full Text].

29. Lindsay TF, Luo XP, Lehotay DC, Rubin BB, Anderson M, Walker PM, Romaschin AD. Ruptured abdominal aortic aneurysm, a "two-hit" ischemia/reperfusion injury: evidence from an analysis of oxidative products. J Vasc Surg 1999; 30: 219-228 [Medline].

30. Wang Q, Doerschuk CM. Neutrophil-induced changes in the biomechanical properties of endothelial cells: roles of ICAM-1 and reactive oxygen species. J Immunol 2000; 164: 6487-6494 [Abstract/Free Full Text].

31. Weitberg AB, Corvese D. Oxygen radicals potentiate the genetic toxicity of tobacco-specific nitrosamines. Clin Genet 1993; 43: 88-91 [Medline].

32. Mota-Filipe H, McDonald MC, Cuzzocrea S, Thiemermann C. A membrane-permeable radical scavenger reduces the organ injury in hemorrhagic shock. Shock 1999; 12: 255-261 [Medline].

33. Ischiropoulos H, al-Mehdi AB, Fisher AB. Reactive species in ischemic rat lung injury: contribution of peroxynitrite. Am J Physiol 1995;269(2 Pt 1):L158-164.

34. Obayashi H, Koshi S, Miyauchi Y, Inoue M. Inhibition of posthemorrhagic transfusion-induced gastric injury by a long-acting superoxide dismutase derivative. Proc Soc Exp Biol Med 1991; 196: 164-169 [Medline].

35. McCord JM. Oxygen-derived free radicals in postischemic tissue injury. N Engl J Med 1985; 312: 159-163 [Abstract].

36. Okayama N, Park JH, Coe L, Granger DN, Ma L, Hisa CJ, Alexander JS. Polynitroxyl alpha-alpha-hemoglobin (PNH) inhibits peroxide and superoxide-mediated neutrophil adherence to human endothelial cells. Free Radic Res 1999; 31: 53-58 [Medline].

37. Russell J, Epstein CJ, Grisham MB, Alexander JS, Yeh KY, Granger DN. Regulation of E-selectin expression in postischemic intestinal microvasculature. Am J Physiol Gastrointest Liver Physiol 2000; 278: G878-885 [Abstract/Free Full Text].

38. Claridge JA, Enelow RI, Young JS. Hemorrhage and resuscitation induce delayed inflammation and pulmonary dysfunction in mice. J Surg Res 2000; 92: 206-213 [Medline].

39. Hierholzer C, Kalff JC, Omert L, Tsukada K, Loeffert JE, Watkins SC, Billiar TR, Tweardy DJ. Inteleukin-6 production in hemorrhagic shock is accompanied by neutrophil recruitment and lung injury. Am J Physiol 1998;275(3 Pt 1):L611-621.

40. Angle N, Hoyt DB, Coimbra R, Liu F, Herdon-Remelius C, Loomis W, Junger WG. Hypertonic saline resuscitation diminishes lung injury by suppressing neutrophil activation after hemorrhagic shock. Shock 1998; 9: 164-170 [Medline].

41. Abraham E, Carmondy A, Shenkar R, Arcaroli R. Neutrophils as early immunologic effectors in hemorrhage or endotoxemia induced acute lung injury. Am J Physiol 2000; 279: L1137-1146 [Abstract/Free Full Text].




This article has been cited by other articles:


Home page
 Am. J. Respir. Crit. Care Med.Home page
T. R. Richens, D. J. Linderman, S. A. Horstmann, C. Lambert, Y.-Q. Xiao, R. L. Keith, D. M. Boe, K. Morimoto, R. P. Bowler, B. J. Day, et al.
Cigarette Smoke Impairs Clearance of Apoptotic Cells through Oxidant-dependent Activation of RhoA
Am. J. Respir. Crit. Care Med., June 1, 2009; 179(11): 1011 - 1021.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Respir. Crit. Care Med.Home page
J. W. Zmijewski, E. Lorne, X. Zhao, Y. Tsuruta, Y. Sha, G. Liu, and E. Abraham
Antiinflammatory Effects of Hydrogen Peroxide in Neutrophil Activation and Acute Lung Injury
Am. J. Respir. Crit. Care Med., April 15, 2009; 179(8): 694 - 704.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Respir. Crit. Care Med.Home page
J. J. Arcaroli, J. E. Hokanson, E. Abraham, M. Geraci, J. R. Murphy, R. P. Bowler, C. A. Dinarello, L. Silveira, J. Sankoff, D. Heyland, et al.
Extracellular Superoxide Dismutase Haplotypes Are Associated with Acute Lung Injury and Mortality
Am. J. Respir. Crit. Care Med., January 15, 2009; 179(2): 105 - 112.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Lung Cell. Mol. Physiol.Home page
B. A. Jacob, K. M. Porter, S. C. Elms, P.-Y. Cheng, D. P. Jones, and R. L. Sutliff
HIV-1-induced pulmonary oxidative and nitrosative stress: exacerbated response to endotoxin administration in HIV-1 transgenic mouse model
Am J Physiol Lung Cell Mol Physiol, October 1, 2006; 291(4): L811 - L819.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Respir. Cell Mol. Bio.Home page
R. J. Tan, J. S. Lee, M. L. Manni, C. L. Fattman, J. M. Tobolewski, M. Zheng, J. K. Kolls, T. R. Martin, and T. D. Oury
Inflammatory Cells as a Source of Airspace Extracellular Superoxide Dismutase after Pulmonary Injury
Am. J. Respir. Cell Mol. Biol., February 1, 2006; 34(2): 226 - 232.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Lung Cell. Mol. Physiol.Home page
J. Y. Kim, J. S. Park, D. Strassheim, I. Douglas, F. Diaz del Valle, K. Asehnoune, S. Mitra, S. H. Kwak, S. Yamada, I. Maruyama, et al.
HMGB1 contributes to the development of acute lung injury after hemorrhage
Am J Physiol Lung Cell Mol Physiol, May 1, 2005; 288(5): L958 - L965.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Respir. Cell Mol. Bio.Home page
R. P. Bowler, M. Nicks, K. Tran, G. Tanner, L.-Y. Chang, S. K. Young, and G. S. Worthen
Extracellular Superoxide Dismutase Attenuates Lipopolysaccharide-Induced Neutrophilic Inflammation
Am. J. Respir. Cell Mol. Biol., October 1, 2004; 31(4): 432 - 439.
[Abstract] [Full Text] [PDF]

Home page
 Eur Respir JHome page
J.D. Crapo
Oxidative stress as an initiator of cytokine release and cell damage
Eur. Respir. J., September 20, 2003; 22(44_suppl): 4s - 6s.
[Full Text] [PDF]

Home page
 Am. J. Physiol. Lung Cell. Mol. Physiol.Home page
R. P. Bowler, J. Arcaroli, E. Abraham, M. Patel, L.-Y. Chang, and J. D. Crapo
Evidence for extracellular superoxide dismutase as a mediator of hemorrhage-induced lung injury
Am J Physiol Lung Cell Mol Physiol, April 1, 2003; 284(4): L680 - L687.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Respir. Crit. Care Med.Home page
R. P. Bowler and J. D. Crapo
Oxidative Stress in Airways: Is There a Role for Extracellular Superoxide Dismutase?
Am. J. Respir. Crit. Care Med., December 15, 2002; 166(12): S38 - 43.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
R. P. Bowler, M. Nicks, D. Aa. Olsen, I. B. Thogersen, Z. Valnickova, P. Hojrup, A. Franzusoff, J. J. Enghild, and J. D. Crapo
Furin Proteolytically Processes the Heparin-binding Region of Extracellular Superoxide Dismutase
J. Biol. Chem., May 3, 2002; 277(19): 16505 - 16511.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Respir. Crit. Care Med.Home page
M. J. TOBIN
Critical Care Medicine in AJRCCM 2001
Am. J. Respir. Crit. Care Med., March 1, 2002; 165(5): 565 - 583.
[Full Text] [PDF]

Home page
 Am. J. Respir. Crit. Care Med.Home page
R. Bowler
ERRATUM: EXTRACELLULAR SUPEROXIDE DISMUTASE ATTENUATES LUNG INJURY AFTER HEMORRHAGE
Am. J. Respir. Crit. Care Med., March 1, 2002; 165(5): 730 - 730.
[Full Text] [PDF]

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow A correction has been published
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by BOWLER, R. P.
Right arrow Articles by ABRAHAM, E.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by BOWLER, R. P.
Right arrow Articles by ABRAHAM, E.

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Proc. Am. Thorac. Soc. Am. J. Respir. Cell Mol. Biol.
Copyright © 2001 American Thoracic Society