Tuberc Respir Dis.  2010 Jun;68(6):334-344. 10.4046/trd.2010.68.6.334.

Moxifloxacin Ameliorates Oleic Acid-induced Acute Lung Injury by Modulation of Neutrophilic Oxidative Stress in Rats

• Kim BY
• Lee YM

• KMID: 1458042
• DOI: http://doi.org/10.4046/trd.2010.68.6.334

Abstract

BACKGROUND
Based on the known immunoregulatory functions of moxifloxacin on phagocytes, the therapeutic effect of moxifloxacin on oleic acid (OA)-induced acute lung injury (ALI) was investigated.
METHODS
Moxifloxacin (10 mg/kg) was given to male Sprague-Dawley rats that had been given oleic acid (OA, 30 microliter) intravenously. Five hours after OA injection, parameters demonstrating ALI were assessed to measure the effects of moxifloxacin on acute lung injury.
RESULTS
The pathological findings of OA-induced ALI's was diminished by moxifloxacin. Through ultrastructural and CeCl3 EM histochemistry, moxifloxacin was confirmed to be effective in decreasing oxidative stress in the lung as well. Indices of ALI, such as lung weight/body weight ratio, protein content in bronchoalveolar lavage fluid, and lung myeloperoxidase were decreased by moxifloxacin. In diaminobenzidine immunohistochemistry, fluorescent immunohistochemistry, and Western blotting of the lung, moxifloxacin had decreased the enhanced expression of secretory phospholipase A2 (sPLA2) by OA.
CONCLUSION
We concluded that moxifloxacin was effective in lessening acute inflammatory pulmonary edema caused by OA, by inhibiting the neutrophilic respiratory burst, which was initiated by the activation of sPLA2.

Keyword

Acute Lung Injury; Neutrophils; Oleic Acid; Free Radicals; Phospholipases A2, Secretory

MeSH Terms

Acute Lung Injury
Animals
Aza Compounds
Blotting, Western
Bronchoalveolar Lavage Fluid
Cerium
Free Radicals
Humans
Immunohistochemistry
Lung
Male
Neutrophils
Oleic Acid
Oxidative Stress
Peroxidase
Phagocytes
Phospholipases A2, Secretory
Pulmonary Edema
Quinolines
Rats
Rats, Sprague-Dawley
Respiratory Burst
Aza Compounds
Cerium
Free Radicals
Oleic Acid
Peroxidase
Phospholipases A2, Secretory
Quinolines

Figure

• 

  Figure 1 (A) The effect of oleic acid on the morphological changes in the lung. Intraalveolar phagocytes, especially neutrophils were found abundantly. Hyaline membrane and wide spread hemorrhage were found also (H&E stain, ×40). (B) The effect of moxifloxacin on the histological changes in the lung of rat given oleic acid intravenously. Alveoli are patent and the accumulation and migration of neutrophils into the alveolar lumen were significantly lessened by moxifloxacin. Generalized inflammatory changes were diminished by moxifloxacin (H&E stain, ×100).

• 

  Figure 2 The ultrastructural changes of alveolar type II cells of the lung in oleic acid (OA), OA with moxifloxacin treated rats. Well preserved lamellar bodies were found in alveolar type II cells of the control lung (A). In OA treated rat, a vacuolization of lamellar bodies of alveolar type II cells was prominent, a direct evidence of oxidative stress (B). In contrast, OA with moxifloxacin treated rats, lamellar bodies of alveolar type II cells were relatively well preserved (C) (Original magnification ×3,000).

• 

  Figure 3 The detection of hydrogen peroxide with CeCl3 cytochemical electro microscopy. In control and oleic acid with moxifloxacin rats, few cerrous perhydroxide granules were found in the vicinity of alveolar type II cells (A, C). In contrast, dense deposits of cerrous perhydroxide granules were found along the membrane of alveolar type II cell (B) (Original magnification ×3,000).

• 

  Figure 4 (A) An immunohistochemistry for the detection of secretory phospholipase A2 (sPLA2) in the normal rat's lung. Staining sPLA2 with diaminobenzidine revealed few reactive cells against sPLA2 antibody. Primary antibody against sPLA2 was goat anti-human sPLA2 polyclonal antibody (dilutional factor ×50) and secondary antibody was biotinylated anti-goat IgG (dilutional factor ×100) (Original magnification ×200). (B) A representative of immunohistochemistry for detection of sPLA2 in the lung of oleic acid (OA) given rat. In the alveoli, abundant migrated neutrophils of which cytoplasm and cell membrane were intensely reacted with diaminobenzidine signifying strong activation of sPLA2 in these cells. Hyaline membranes were found also in the alveolar lumen. Primary antibody against sPLA2 was goat anti-human sPLA2 polyclonal antibody (dilutional factor ×50) and secondary antibody was biotinylated anti-goat IgG (dilutional factor ×100) (Original magnification ×200). (C) An immunohistochemical photograph of detecting sPLA2 in the lung of rat given OA and moxifloxacin. Few phagocytes were found in the lung and diaminobenzidine positive cell was not found. Primary antibody against sPLA2 was goat anti-human sPLA2 polyclonal antibody (dilutional factor ×50) and secondary antibody was biotinylated anti-goat IgG (dilutional factor ×100) (Original magnification ×200).

• 

  Figure 5 (A) A fluorescent immunohistochemistry for detection of secretory phospholipase A2 (sPLA2) in the normal rat's lung. Counter-staining with propidium iodide of sPLA2 revealed few cells of sPLA2 positive against sPLA2 antibody. Primary antibody against sPLA2 was goat anti-human sPLA2 polyclonal antibody (dilutional factor ×50) and secondary antibody was biotinylated anti-goat IgG (dilutional factor ×100) (Original magnification ×200). (B) A representative of fluorescent immunohistochemistry for detection of sPLA2 in the lung of oleic acid (OA) given rat. Dense, highly fluorescent area were found in the lung depicting the strong expression of PLA2 in the lung. Primary antibody against sPLA2 was goat anti-human sPLA2 polyclonal antibody (dilutional factor ×50) and secondary antibody was biotinylated anti-goat IgG (dilutional factor ×100) (Original magnification ×200). (C) A fluorescent immunohistochemical photograph of detecting sPLA2 in the lung of rat given OA and moxifloxacin. Fluorescent area of lung by OA was strikingly diminished by moxifloxacin signifying the inhibition of PLA2. Primary antibody against sPLA2 was goat anti-human sPLA2 polyclonal antibody (dilutional factor ×50) and secondary antibody was biotinylated anti-goat IgG (dilutional factor ×100) (Original magnification ×200).

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  Figure 6 Western blot analysis of secretory phospholipase A2 (sPLA2) in the lung. Oleic acid (OA) treatment increased the expression of sPLA2 but this increased expression was down-regulated by moxifloxacin (MO).

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