Title Page
Contents
I. Introduction 8
II. Materials and Methods 11
Animal model 11
Tissue preparation 12
Lung morphometric analysis 12
TUNEL assay 13
Myeloperoxidase activity 14
Semiquantitative reverse transcriptase - polymerase chain reaction (RT-PCR) of cytokines 14
Western blot of p67phox membrane translocation(이미지참조) 16
Statistics 16
III. Results 20
Survival rate and body weight gain 20
WBC count in blood 20
Morphometric analysis 20
TUNEL assay 21
MPO activity 21
mRNA expression of pro-inflammatory cytokines 22
Oxidative stress in lung tissue 22
IV. Discussion 33
V. References 39
Abstract 45
Abstract 47
Table 1. The following sequences of primers for tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), transforming growth factor-beta (TGF-β) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were used. 19
Figure 1. Ratpups of hyperoxia group (HC and HG) were kept with dam in Plexiglas chambers in which the oxygen concentration of 90% was maintained. Humidity and environmental temperature were maintained at 50% and 24°C, respectively. Nursing mother rats were rotated between... 18
Figure 2. (A) Kaplan-Meier curve showing decreased survival in hyperoxia (HC), and improved survival with G-CSF treatment (HG), but there was no statistical significance. (B) Hyperoxia exposed rats (HC) show significantly retarded weight gain compared to normoxia (NC, NG), and this retarded... 23
Figure 3. Exposure to hyperoxia (HC) significantly increases WBC in blood. G-CSF treatment decreases WBC in blood, but there was no statistical significance. 24
Figure 4. (A) Lung structure of neonatal rats in normoxia at day 14 (NC); (B) G-CSF treatment to normoxia group did not result in any changes in day 14 (NG); (C) hyperoxia results in impaired alveolarization, or fewer and larger alveoli and simplified alveolar architecture (HC); (D) G-CSF... 25
Figure 5. (A) mean linear intercept; (B) mean alveolar volume Hyperoxia exposed lungs (HC) show significant increase in mean linear intercept (MLI) and mean alveolar volume. G-CSF treatment (HG) improves hyperoxia-induced morphometric abnormalities. 26
Figure 6. (A) Hyperoxia exposed lungs (HC) show significant increase in TUNEL positive cells, and this increase is attenuated by G-CSF treatment (HG). (B) Fluorescent green spots mean TUNEL positive cells with apoptotic nuclei on fluorescent microscope (TUNEL staining, magnification... 28
Figure 7. Hyperoxia exposed lungs (HC) show significant increase in MPO activity, and this increase is attenuated by G-CSF treatment (HG). 30
Figure 8. Hyperoxia exposed lungs (HC) show significant increase in mRNA expression of TNF-α, IL-6 and TGF-β. G-CSF treatment attenuated TGF-β expression at day 14. 31
Figure 9. Hyperoxia exposed lungs (HC) show significant increase increase in p67phox as an indicator of oxidative stress both in cytosol and membrane. G-CSF treatment (HG) decreased p67phox membrane translocation at day 14.(이미지참조) 32