Journal List > Tuberc Respir Dis > v.69(4) > 1001541

TOOLS
Lee: Moxifloxacin Alleviates Oleic Acid-provoked Neutrophilic Respiratory Burst in the Rat Lung through the Inhibition of Cytosolic Phospholipase A2
Original Article
Tuberculosis and Respiratory Diseases

Tuberculosis and Respiratory Diseases 2010; 69(4): 256-264.

Published online: 31 October 2010

DOI: https://doi.org/10.4046/trd.2010.69.4.256

Moxifloxacin Alleviates Oleic Acid-provoked Neutrophilic Respiratory Burst in the Rat Lung through the Inhibition of Cytosolic Phospholipase A2

Young Man Lee, M.D.

Department of Physiology, Daegu Catholic University School of Medicine, Daegu, Korea.

Address for correspondence: Young Man Lee, M.D. Department of Physiology, Daegu Catholic University School of Medicine, 3056-6, Daemyung 4-dong, Nam-gu, Daegu 705-718, Korea. Phone: 82-53-650-4472, Fax: 82-53-621-4106, leeym@cu.ac.kr

Received 5 August 2010       Accepted 16 September 2010

Copyright © 2010. The Korean Academy of Tuberculosis and Respiratory Diseases. All rights reserved.

Abstract

Background

According to the notion of the immunoregulatory functions of moxifloxacin (MFX), the effect of MFX on the neutrophilic respiratory burst in conjunction with the expression of cytosolic phospholipase A2 (cPLA2) was investigated.

Methods

The effects and possible mechanisms of MFX on neutrophilic respiratory burst in oleic acid (OA)-induced acutely injured rats lung and OA-stimulated, isolated murine neutrophils were probed, associated with the expression of cytosolic phospholipase A2 in vivo and in vitro.

Results

In the OA-induced acutely-injured lungs, neutrophils were accumulated, which was attenuated by MFX. The parameters denoting a neutrophilic respiratory burst, such as nitro blue tetrazolium reaction, cytochrome-c reduction, neutrophil aggregation, H2O2 production in neutrophils revealed increased neutrophilic respiratory burst by OA, and MFX decreased all of these parameters. In addition, the enhanced expression of cPLA2 in the lung and isolated murine neutrophils by OA were decreased by MFX.

Conclusion

MFX suppresses the OA-induced neutrophilic respiratory burst by the suppression of cPLA2 in neutrophils.

Keywords: Oleic Acid, Neutrophils, Phospholipases A2, Cytosolic, Moxifloxacin

Figures and Tables

Figure 1
Histological findings of the lung in sham, oleic acid (OA) and moxifloxacin (MFX)-group. In sham rat, alveolar septa and alveolar lumen were well preserved. No inflammatory cell was noted (A, H&E stain, ×100). In contrast, the lung of OA-group showed migrated phagocytes and red blood cells in the alveolar lumen denoting acute inflammatory reaction. Hyaline membranes were shown also (B, H&E stain, ×40). In MFX group, the alveolar lumen was patent and the inflammatory reaction was minimal even if remnants of hyaline membrane and monocytes were noted (C, H&E stain, ×100).
trd-69-256-g001
Figure 2
The representative of findings of the nitro blue tetrazolium (NBT) test in cytospinned bronchalveolar lavage fluid (BALF). In the neutrophil from the BALF of sham group, the resting, normal neutrophil showed no formazan granule in the cytoplasm, denoting absence of respiratory burst (A). In contrast, in BALF of oleic acid-group, neutrophils were clumped and deep, dense formazan granules were found in the cytoplasms of activated neutrophils, depicting severe respiratory burst (B). In neutrophils from the BALF of moxifloxacin group, formazan granules were not found suggesting the abscense or minimal respiratory burst (C).
trd-69-256-g002
Figure 3
Cerium chloride electron microscopic cytochemistry for the detection of hydrogen peroxide in the infilterated neutrophils in the lung. In sham group, cerrous perhydroxide granules was not found in the vicinity neutrophil (A, arrow heads, ×6,000). On the contrary, in the lung of oleic acid-group, dense deposits of cerrous perhydroxide granules were found along the neutrophilic membrane (B, arrow heads, ×6,000). In moxifloxacin group, cerrous perhydroxide granules were found but the deposits were slight comparing with those of oleic acid-group's (C, arrow heads, ×6,000). pmn: polymorphonuclear neutrophil.
trd-69-256-g003
Figure 4
Immunohistochemistry of the cells in bronchalveolar lavage fluid (BALF). Cells from the BALF of sham group were almost monocytes which did not express cytololic phospholipase A2 (cPLA2) (A, ×100). In contrast, the cells from the BALF of oleic acid-group were almost neutrophils which expressed cPLA2 conspicuously in their cytoplasms as bright dots (B, ×100). In BALF of moxifloxacin-group, neutrophils and monocytes were intermingled but monocytes were predominant, and the expression of cPLA2 was not observed (C, ×100).
trd-69-256-g004
Figure 5
Immunoprecipitation of cytololic phospholipase A2 (cPLA2) in isolated murine neutrophils. Oleic acid (OA) increased the expression of cPLA2 compared with that of control neutrophils. Moxifloxacin (MFX) suppressed the expression of cPLA2 caused by OA.
trd-69-256-g005
Table 1
Effect of oleic acid and moxifloxacin on the infiltration and migration of neutrophils in the lung
trd-69-256-i001

Values are given as mean±SE. n indicates number of experiments.

OA: oleic acid; MFX: moxifloxacin; MPO: myeloperoxidase; BAL: bronchoalveolar lavage; PMN: polymorphonuclear neutrophils.

*p<0.001, sham vs. OA, †p<0.001, OA vs. OA+MFX.

Table 2
Effect of oleic acid and moxifloxacin on the neutrophilic oxidative stress and chemotaxis
trd-69-256-i002

Values are given as mean±SE. n indicates number of experiments.

OA: oleic acid; MFX: moxifloxacin; PMN: polymorphonuclear neutrophil.

*p<0.001 sham vs. OA, †positive, ‡p<0.01 OA vs. OA+MFX.

References

1. Ware LB. Advances in the pathogenesis and treatment of the acute respiratory distress syndrome. Clin Pul Med. 2003. 10:208–218.
2. Kitsiouli E, Nakos G, Lekka ME. Phospholipase A2 subclasses in acute respiratory distress syndrome. Biochim Biophys Acta. 2009. 1792:941–953.
3. Nakos G, Kitsiouli E, Hatzidaki E, Koulouras V, Touqui L, Lekka ME. Phospholipases A2 and platelet-activating-factor acetylhydrolase in patients with acute respiratory distress syndrome. Crit Care Med. 2005. 33:772–779.
4. Stommer P, Steinmann U. Phospholipase A2 induced diffuse alveolar damage: effect of indomethacin and dexamethasone upon morphology and plasma-histamine level. Klin Wochenschr. 1989. 67:171–176.
5. Schaloske RH, Dennis EA. The phospholipase A2 superfamily and its group numbering system. Biochim Biophys Acta. 2006. 1761:1246–1259.
6. Wheeler DS. Phospholipase A2 and acute lung injury: it's just not that simple. Crit Care Med. 2005. 33:904–905.
7. Pruzanski W, Vadas P. Phospholipase A2: a mediator between proximal and distal effectors of inflammation. Immunol Today. 1991. 12:143–146.
8. Arbibe L, Vial D, Touqui L. Phospholipase A2 and acute respiratory distress syndrome. Prog Surg. 1997. 24:79–87.
9. Kramer RM, Sharp JD. Structure, function and regulation of Ca2+-sensitive cytosolic phospholipase A2 (cPLA2). FEBS Lett. 1997. 410:49–53.
10. Dana R, Malech HL, Levy R. The requirement for phospholipase A2 for activation of the assembled NADPH oxidase in human neutrophils. Biochem J. 1994. 297:217–223.
11. Lucas KK, Dennis EA. The ABC's of Group IV cytosolic phospholipase A2. Biochim Biophys Acta. 2004. 1636:213–218.
12. Huang HC, Shieh CC, Yu WL, Cheng KC, Chen CC, Chang ST, et al. Comparing the protective effects of ciprofloxacin, moxifloxacin and levofloxacin in mice with lipopolysaccharide-induced acute lung injuries. Respirology. 2008. 13:47–52.
13. Wang JP, Raung SL, Huang LJ, Kuo SC. Involvement of cyclic AMP generation in the inhibition of respiratory burst by 2-phenyl-4-quinolone (YT-1) in rat neutrophils. Biochem Pharmacol. 1998. 56:1505–1514.
14. Weiner RE, Sasso DE, Gionfriddo MA, Syrbu SI, Smilowitz HM, Vento J, et al. Early detection of bleomycin-induced lung injury in rat using indium-111-labeled antibody directed against intercellular adhesion molecule-1. J Nucl Med. 1998. 39:723–728.
15. Goldblum SE, Wu KM, Jay M. Lung myeloperoxidase as a measure of pulmonary leukostasis in rabbits. J Appl Physiol. 1985. 59:1978–1985.
16. Hobson J, Wright J, Churg A. Histochemical evidence for generation of active oxygen species on the apical surface of cigarette-smoke-exposed tracheal explants. Am J Pathol. 1991. 139:573–580.
17. Haslett C, Guthrie LA, Kopaniak MM, Johnston RB Jr, Henson PM. Modulation of multiple neutrophil functions by preparative methods or trace concentrations of bacterial lipopolysaccharide. Am J Pathol. 1985. 119:101–110.
18. Botha AJ, Moore FA, Moore EE, Fontes B, Banerjee A, Peterson VM. Postinjury neutrophil priming and activation states: therapeutic challenges. Shock. 1995. 3:157–166.
19. Dalhoff A, Shalit I. Immunomodulatory effects of quinolones. Lancet Infect Dis. 2003. 3:359–371.
20. Koksel O, Cinel I, Tamer L, Cinel L, Ozdulger A, Kanik A, et al. N-acetylcysteine inhibits peroxynitrite-mediated damage in oleic acid-induced lung injury. Pulm Pharmacol Ther. 2004. 17:263–270.
21. Karagiorga G, Nakos G, Galiatsou E, Lekka ME. Biochemical parameters of bronchoalveolar lavage fluid in fat embolism. Intensive Care Med. 2006. 32:116–123.
22. Julien M, Hoeffel JM, Flick MR. Oleic acid lung injury in sheep. J Appl Physiol. 1986. 60:433–440.
23. Lee YM, Kim BY, Park YY. Role of the PLA2-activated neutrophilic oxidative stress in oleic acid-induced acute lung injury. Tuberc Respir Dis. 2010. 68:55–61.
24. Shmelzer Z, Haddad N, Admon E, Pessach I, Leto TL, Eitan-Hazan Z, et al. Unique targeting of cytosolic phospholipase A2 to plasma membranes mediated by the NADPH oxidase in phagocytes. J Cell Biol. 2003. 162:683–692.
25. Leslie CC. Properties and regulation of cytosolic phospholipase A2. J Biol Chem. 1997. 272:16709–16712.
26. Ghosh M, Tucker DE, Burchett SA, Leslie CC. Properties of the Group IV phospholipase A2 family. Prog Lipid Res. 2006. 45:487–510.
27. Nagase T, Uozumi N, Ishii S, Kume K, Izumi T, Ouchi Y, et al. Acute lung injury by sepsis and acid aspiration: a key role for cytosolic phospholipase A2. Nat Immunol. 2000. 1:42–46.
28. Magrioti V, Kokotos G. Phospholipase A2 inhibitors as potential therapeutic agents for the treatment of inflammatory diseases. Expert Opin Ther Pat. 2010. 20:1–18.
TOOLS
Similar articles