Tuberc Respir Dis.  2008 Dec;65(6):464-470.

Presumptive Role of Neutrophilic Oxidative Stress in Oxygen-induced Acute Lung Injury in Rats

• Moon Y
• Kim J
• Lee YM

Abstract

BACKGROUND: This study examined the role of neutrophilc oxidative stress in an O2-induced acute lung injury (ALI).
METHODS
For 48 h, experimental rats were exposed to pure oxygen (normobaric hyperoxia) in a plastic cage. Forty-eight hours after O2 breathing, the rats were sacrificed and the parameters for ALI associated with neutrophilic oxidative stress were assessed.
RESULTS
Normobaric pure oxygen induced ALI, which was quite similar to ARDS. The O2-induced neutrophilic oxidative stress was identified by confirming of the increase in lung myeloperoxidase, BAL neutrophils, malondialdehyde (MDA), cytosolic phospholipase A2 (cPLA2) activity in the lung, histological changes and BAL cytospin morphology.
CONCLUSION
In part, ALI-caused by oxygen is affected by neutrophils especially by the generation of free radicals.

Keyword

Acute lung injury; O2 toxicity; Neutrophils

MeSH Terms

Acute Lung Injury
Animals
Cytosol
Free Radicals
Lung
Malondialdehyde
Neutrophils
Oxidative Stress
Oxygen
Peroxidase
Phospholipases A2
Plastics
Rats
Respiration
Free Radicals
Malondialdehyde
Oxygen
Peroxidase
Phospholipases A2
Plastics

Figure

• 

  Figure 1 (A) The representative photograph of BALF cytospin of rat given ambient air. Almost all of the cells are monocytes (Wright's stain, ×100). (B) The representative photograph of BALF cytospin of rats given 100% O2 for 48 h. The intermingling of monocytes and neutrophils is noted. Neutrophils are much more abundant than monocytes (Wright's stain, ×40).

• 

  Figure 2 (A) The normal histological finding of the lung of rat given ambient air. Patent alveoli and thin, normal alveolar septa are noted (H&E stain, ×100). (B) The histological finding of the lung given 100% O2 for 48 h. Diffuse hemorrhage is noted. Intraalveolar hemorrhage, migrated phagocytes and hyaline membranes are shown (H&E stain, ×40). (C) The photograph of the migration of phagocytes from vascular lumen into alveolar lumen in the lung of rats given 100% O2 for 48 h. Phagocytes including neutrophils are trapped in the perivascular cuff (below right) and phagocytes are migrating into alveolar lumen from adjacent vessel. Extravasation of the red blood cells are noted also (H&E stain, ×100). (D) The magnification of Figure C. Note the migration of phagocytes from vessel into alveolar lumen. Neutrophils are moving into the alveolar lumen by a diapedesis and red blood cells are migrating into the alveolar lumen also (H&E stain, ×400).

Reference

1. Hay WW Jr, Bell EF. Oxygen therapy, oxygen toxicity, and STOP-ROP trial. Pediatrics. 2000. 105:424–425.

2. Crapo JD, Barry BE, Foscue HA, Shelburne J. Structural and biochemical changes in rat lungs occurring during exposures to lethal and adaptive doses of oxygen. Am Rev Respir Dis. 1980. 122:123–143.

3. Chabot F, Mitchell JA, Gutteridge JM, Evans TW. Reactive oxygen species in acute lung injury. Eur Respir J. 1998. 11:745–757.

4. Guidot DM, Repine JE, Kitlowski AD, Flores SC, Nelson SK, Wright RM, et al. Mitochondrial respiration scavenges extramitochondrial superoxide anion via a nonenzymatic mechanism. J Clin Invest. 1995. 96:1131–1136.

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

6. Brown RE, Jarvis KL, Hyland KJ. Protein measurement using bicinchoninic acid: elimination of interfering substances. Anal Biochem. 1989. 180:136–139.

7. Goldblum SE, Wu KM, Jay M. Lung myeloperoxidase as a measure of pulmonary leukostasis in rabbits. J Appl Physiol. 1985. 59:1978–1985.

8. Carraway MS, Piantadosi CA, Jenkinson CP, Huang YC. Differential expression of arginase and iNOS in the lung in sepsis. Exp Lung Res. 1998. 24:253–268.

9. Katsumata M, Gupta C, Goldman AS. A rapid assay for activity of phospholipase A2 using radioactive substrate. Anal Biochem. 1986. 154:676–681.

10. Capellier G, Maupoil V, Boussat S, Laurent E, Neidhardt A. Oxygen toxicity and tolerance. Minerva Anestesiol. 1999. 65:388–392.

11. Torbati D, Tan GH, Smith S, Frazier KS, Gelvez J, Fakioglu H, et al. Multiple-organ effect of normobaric hyperoxia in neonatal rats. J Crit Care. 2006. 21:85–93.

12. Roman K, Vladimíra M, Alexander C. Highly specific, simple and rapid method for the determination of malondialdehyde in blood using high-performance liquid chromatography. Clin Chem Lab Med. 2002. 40:1032–1035.

13. Sukhotnik I, Coran AG, Greenblatt R, Brod V, Mogilner J, Shiloni E, et al. Effect of 100% oxygen on E-selectin expression, recruitment of neutrophils and enterocyte apoptosis following intestinal ischemia-reperfusion in a rat. Pediatr Surg Int. 2008. 24:29–35.

14. Shvedova AA, Kisin ER, Murray AR, Kommineni C, Castranova V, Fadeel B, et al. Increased accumulation of neutrophils and decreased fibrosis in the lung of NADPH oxidase-deficient C57BL/6 mice exposed to carbon nanotubes. Toxicol Appl Pharmacol. 2008. 231:235–240.

15. Shasby DM, Fox RB, Harada RN, Repine JE. Reduction of the edema of acute hyperoxic lung injury by granulocyte depletion. J Appl Physiol. 1982. 52:1237–1244.

16. Hybertson BM, Lee YM, Repine JE. Phagocytes and acute lung injury: dual roles for interleukin-1. Ann N Y Acad Sci. 1997. 832:266–273.

17. Lin Y, Jamieson D. Role of neutrophils in hyperbaric and normobaric oxygen toxicity. Pathophysiology. 1995. 2:9–16.

18. Prudhomme JB, Ware LB. Acute lung injury and acute respiratory distress syndrome: mechanisms and potential new therapies. Drug Discov Today Dis Mech. 2004. 1:123–128.

19. Lee YM, Hybertson BM, Terada LS, Repine AJ, Cho HG, Repine JE. Mepacrine decreases lung leak in rats given interleukin-1 intratracheally. Am J Respir Crit Care Med. 1997. 155:1624–1628.

20. Guidot DM. Endotoxin treatment increases lung mitochondrial scavenging of extramitochondrial superoxide in hyperoxia-exposed rats. Arch Biochem Biophys. 1996. 326:266–270.

21. Boyer CS, Bannenberg GL, Neve EP, Ryrfeldt A, Moldéus P. Evidence for the activation of the signal-responsive phospholipase A2 by exogenous hydrogen peroxide. Biochem Pharmacol. 1995. 50:753–761.