Tuberc Respir Dis.  2011 Aug;71(2):97-105. 10.4046/trd.2011.71.2.97.

A Study on the Mechanism of Immunomodulating Effects of Moxifloxacin in Oleic Acid-Induced Acute Lung Injury

• Lee YM

• KMID: 1846416
• DOI: http://doi.org/10.4046/trd.2011.71.2.97

Abstract

BACKGROUND
It was hypothesized that the immunomodulating effects of moxifloxacin contribute to ameliorate oleic acid (OA)-induced acute lung injury (ALI) by suppression of cytosolic phospholipase A2 (cPLA2). This was based on observations from experiments on rats associated with neutrophilic respiratory burst, cPLA2 activity, and expressions of cPLA2, TNFalpha, and COX-II in the lung.
METHODS
ALI was induced by intravenous injection of OA in male Sprague-Dawley rats. Five hours after OA injection, protein content in bronchoalveolar lavage (BAL), lung myeloperoxidase (MPO) activity, and numbers of BAL neutrophils were measured. As an index of oxidative stress-induced lung injury, the content of malondialdehyde (MDA) in lung tissues was also determined. Lung histology, immunohistochemistry and determination of activity of cPLA2 in lung tissues were carried out. In addition, Western blotting of TNFalpha and COX-II in lung tissues was performed.
RESULTS
The accumulation of neutrophils in the lungs was observed after OA injection. BAL protein was increased along with neutrophilic infiltration and migration by OA. Moxifloxacin decreased all of these parameters of ALI and ameliorated ALI histologically. The increased malondialdehyde (MDA) in the lung by OA was also decreased by moxifloxacin. Moxifloxacin not only suppressed cPLA2 expression in the lungs and neutrophils but also decreased cPLA2 activity in lung tissues of rats given OA. The enhanced expressions of TNFalpha and COX-2 in the lung tissues of rats given OA were also suppressed by moxifloxacin.
CONCLUSION
Moxifloxacin inhibited cPLA2 and down-regulated TNFalpha and COX-2 in the lungs of rats given OA, which resulted in the attenuation of inflammatory lung injury.

Keyword

Moxifloxacin; Oleic Acid; Neutrophils; Acute Lung Injury

MeSH Terms

Acute Lung Injury
Animals
Aza Compounds
Blotting, Western
Bronchoalveolar Lavage
Cytosol
Humans
Immunohistochemistry
Injections, Intravenous
Lung
Lung Injury
Male
Malondialdehyde
Neutrophils
Oleic Acid
Peroxidase
Phospholipases A2
Quinolines
Rats
Rats, Sprague-Dawley
Respiratory Burst
Tumor Necrosis Factor-alpha
Aza Compounds
Malondialdehyde
Oleic Acid
Peroxidase
Phospholipases A2
Quinolines
Tumor Necrosis Factor-alpha

Figure

• 

  Figure 1 Effects of moxifloxacin on histopathology in the lung. (A) Patent alveoli and normal septa of alveoli were well preserved and no phagocyte was found in sham-treated rats (H&E stain, ×40). (B) In contrast, in rats given OA, phagocytes, mainly neutrophils, and red blood cells were present in alveoli. Perivascular cuffing, hyaline membrane, and edematous alveolar septa were evident (H&E stain, ×100). (C) By the administration of moxifloxacin, these pathological findings were mitigated. Even if slight migration of phagocytes was noted, neutrophils were difficult to find. Alveoli were relatively patent and septal edema was not prominent compared with that of rats given OA (H&E stain, ×100). OA: oleic acid.

• 

  Figure 2 Immunohistochemical staining of cPLA2 in the lung. (A) Note the absence of phagocytic cells in alveoli in sham-treated rats (DAB staining, ×100). (B) By contrast, in rats given OA, DAB-stained neutrophils and monocytes were present in alveoli. In the cytoplasm of neutrophils, cPLA2s were well stained and visualized (DAB staining, ×200). (C) In moxifloxacin-treated rats, there was almost no difference in cPLA2 expression comparing with that of sham-treated rats (DAB staining, ×200). cPLA2: cytosolic phospholipase A2; OA: oleic acid.

• 

  Figure 3 Fluorescent immunohistochemistry of lung tissues. (A) The staining of cPLA2 in the lung tissue was not found in sham-treated rats (propidium iodide staining, ×100). (B) By contrast, in rats given OA, cPLA2 was expressed as bright fluorescent area in the lung (propidium iodide staining, ×100). (C) Moxifloxacin suppressed cPLA2 expression in the lung almost completely in rats given OA (propidium iodide staining, ×100). cPLA2: cytosolic phospholipase A2; OA: oleic acid.

• 

  Figure 4 Western blot of TNFα and COX-2 in the lung tissue. The OA increased the expression of TNFα and COX-2. Moxifloxacin effectively suppressed the expression of TNFα and COX-2 which had been upregulated by OA. OA: oleic acid; MO: moxifloxacin.

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