[The effect on anti oxidative damage with aspirin pretreatment in primary cultured rat type II alveolar epithelial cell].

OBJECTIVE: To investigate the protective effect of aspirin on primary cultured type II alveolar epithelial cell (AEC II), and the mechanism of its effect on anti oxidation damage. METHODS: The original generation of adult rat AEC II were cultured and purified. They were divided into normal saline (NS) group, hydrogen peroxide injury group (H(2)O(2) group), and 1, 2, 3 aspirin pretreatment groups (A1-3 groups).In H(2)O(2) group, 0.5 mmol/L H(2)O(2) was added to AEC II after 40 hours of culture to reproduce a cell oxidative injury model. In NS group, only NS was added to AEC II culture. To the A1-3 groups aspirin 50, 100 and 200 μmol/L were added respectively. Cell form, cell count and cell survival rate were observed at 3 hours after H(2)O(2) was given . Immunohistochemical and polymerase chain reaction (PCR) methods were used for the determination of heme oxygenase-1 (HO-1) protein and HO-1 mRNA (20, 40, 60 hours of culture). RESULTS: With trypsin digestion and immune adherence method AEC II could be harvested (2.0-2.5)× , and the purity and activity were both over 90%. Compared with NS group, gaps between cells were widened in H(2)O(2) group, cell account was reduced, and the survival rate (A value) was reduced significantly (0.054 6±0.004 0 vs. 0.103 8±0.009 9, P<0.01). Compared with H(2)O(2) group, in A1-3 groups the number of adherent cells was increased, cell morphology was intact, and no obvious cell shrinkage was found. Higher survival rate (A value) was found in A1-3 groups than that of H(2)O(2) group (0.066 9±0.003 9, 0.071 0±0.006 5, 0.078 7±0.009 2 vs. 0.054 6±0.004 0, all P<0.01). Compared with NS group, HO-1 protein and HO-1 mRNA expression in AEC II after 20, 40 and 60 hours of culture reached peak level at 60 hours, and they were increased significantly in A1-3 groups [protein (A value): 1.59±0.12, 1.60±0.09, 1.61±0.08 vs. 1.25±0.11; mRNA (the ratio of Ct value: 24.31±1.74, 30.45± 2.53, 32.63±3.74 vs. 22.99±1.95, all P<0.05]. There was no significant difference in HO-1 protein expression among A1-3 groups. CONCLUSION: There are significant protective effects of aspirin against anti oxidative damage in cultured AEC II cell. As expression of HO-1 is increased in aspirin groups, it may be considered as a protective factor against anti oxidative damage in AEC II cell culture.