Fig 1.
Effect of probucol on BALF and the histological findings in hyperoxic lung injury.
(A) The results of cell count and level of IL-6 in BALF. Each bar shows mean ± SEM of the ten mice in each group. (B-E) Hematoxylin and eosin staining. In room-air-exposed mice without probucol treatment (B) and room-air-exposed mice treated with probucol (C), no abnormalities were observed. In the lung tissue of hyperoxia-exposed mice without probucol treatment (D), invasion of inflammatory cells, pulmonary edema, and alveolar hemorrhage were observed. In hyperoxia-exposed mice treated with probucol (E), those findings were improved. In the pathological grading, each circle corresponds to one mouse (F). Original magnifications: ×200. **P < 0.01, *p < 0.05.
Fig 2.
Probucol ameliorates oxidative stress and apoptosis in hyperoxic lung injury.
(A-D) The results of immunohistochemistry for 8-OHdG. (A) In room-air-exposed mice treated with probucol, expression of 8-OHdG was not detected. (B) In hyperoxia-exposed mice without probucol treatment, 8-OHdG was strongly expressed in nuclei of lung epithelial cells due to the tissue damage. (C) In hyperoxia-exposed mice treated with probucol, signal of 8-OHdG was decreased. (D) The immunostaining grade for 8-OHdG was significantly decreased by probucol treatment. Original magnifications: ×200. Data are shown as the mean ± SEM from four mice in each group. (E) Western blot analysis for p47phox. Each lane corresponds to the data from one mouse. (F) Relative band intensities from western blot analysis. Optical density values for each individual band were normalized to β-tubulin expression from the same tissue. Data are means ± SEM from three mice. **p < 0.01.
Fig 3.
Probucol ameliorates apoptosis in hyperoxic lung injury.
(A-D) The effect of probucol on TUNEL staining. (A) No positive signals for TUNEL were observed in lung tissues of room-air-exposed mice treated with probucol. (B) There were some TUNEL-positive cells (arrows in F) in the lung tissues of hyperoxia-exposed mice. (C) Probucol treatment decreased the number of TUNEL-positive cells in hyperoxia-exposed mice. (D) Quantitative result of the number of TUNEL-positive cells in lung tissues. Original magnifications: ×200. Data are shown as the mean ± SEM from four mice in each group. **p < 0.01.
Fig 4.
Probucol treatment increases Bcl-XL expression in the lung tissues.
(A) There was no detectable expression of Bcl-XL in room-air-exposed mice (A). In both room-air-exposed mice and hyperoxia-exposed mice, probucol treatment increased Bcl-XL expression in lung epithelial cells (B, D). In hyperoxia-exposed mice without probucol treatment, there was a low-level expression of Bcl-XL (C). Original magnifications: ×200.
Fig 5.
Western blot analysis for p47phox, phospho-ERK1/2, NF-κB (p65), Bcl-XL, Bax (6A7), cleaved caspase-9, and β-tubulin.
(A) Each lane corresponds to the data from one mouse. (B) Relative band intensities from western blot analysis. Optical density values for each individual band were normalized to β-tubulin expression from the same tissue. Data are means ± SEM from three mice. **p < 0.01.